SHIGA TOXIN A SUBUNIT EFFECTOR POLYPEPTIDES, SHIGA TOXIN EFFECTOR SCAFFOLDS, AND CELL-TARGETING MOLECULES FOR SITE-SPECIFIC CONJUGATION
20240082404 ยท 2024-03-14
Inventors
Cpc classification
A61K45/06
HUMAN NECESSITIES
A61K38/04
HUMAN NECESSITIES
A61K2039/6037
HUMAN NECESSITIES
A61K47/6829
HUMAN NECESSITIES
C07K2319/55
CHEMISTRY; METALLURGY
A61P35/00
HUMAN NECESSITIES
International classification
A61K45/06
HUMAN NECESSITIES
A61K47/68
HUMAN NECESSITIES
A61K38/16
HUMAN NECESSITIES
Abstract
The present invention provides Shiga toxin A Subunit derived polypeptides, scaffolds, and cell-targeting molecules comprising amino acid substitutions which equip the molecules with site-specific positions (and often unique amino acid residues in the molecule) for linking other molecules while retaining Shiga toxin function(s), such as, e.g., efficient intracellular routing and/or potent cytotoxicity. The present invention also provides cell-targeting molecules, and/or components thereof, which comprise site-specific positions for linking other molecules, such as, e.g., agents that alters a property of the cell-targeting molecule or a cargo for delivery. Certain molecules comprising a polypeptide of the present invention exhibit reduced immunogenicity and/or are well-tolerated by mammals. The cell-targeting molecules of the present invention, and compositions thereof, have uses, e.g., for the selective delivery of cargos to target-expressing cells and as diagnostic and/or therapeutic molecules for the treatment of a variety of diseases, disorders, and conditions, which include genetic disorders, genetic predispositions, infections, cancers, tumors, growth abnormalities, and/or immune disorders.
Claims
1. A method of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a cell-targeting molecule comprising: (a) a Shiga toxin effector polypeptide; (b) a binding region capable of specifically binding at least one extracellular target biomolecule; and (c) a linker peptide between the Shiga toxin effector polypeptide and the binding region, wherein the Shiga toxin effector polypeptide comprises an amino acid sequence having at least 90% sequence identity to amino acids 1 to 251 of SEQ ID NO: 1, wherein the Shiga toxin effector polypeptide comprises amino acid substitutions S45I, V54I, R55L, I57F, P59F, E60T, E61L, G110A, and R188A in SEQ ID NO: 1, wherein the Shiga toxin effector polypeptide comprises one or more cysteines suitable for conjugation to a heterologous molecule, wherein the Shiga toxin effector polypeptide comprises at least one of the following amino acid substitutions in SEQ ID NO: 1: S8C, S16C, S22C, S43C, S146C, and S186C; or a pharmaceutical composition thereof, wherein treating the cancer comprises selectively killing target cells that contribute to the cancer; and wherein the target cells express the extracellular target biomolecule.
2. The method of claim 1, wherein the Shiga toxin effector polypeptide comprises an asparagine at the amino acid residue corresponding to position 75 of SEQ ID NO: 1, a tyrosine at the amino acid residue corresponding to position 114 of SEQ ID NO: 1, an arginine at the amino acid residue corresponding to position 170 of SEQ ID NO: 1, an arginine at the amino acid residue corresponding to position 176 of SEQ ID NO: 1, and a tryptophan at the amino acid residue corresponding to position 203 of SEQ ID NO: 1.
3. The method of claim 1, wherein the Shiga toxin effector polypeptide comprises the amino acid substitutions R248A and R251A in SEQ ID NO: 1.
4. The method of claim 1, wherein the Shiga toxin effector polypeptide comprises the amino acid substitution C242S in SEQ ID NO: 1.
5. The method of claim 1, wherein the Shiga toxin effector polypeptide comprises at least one of the amino acid substitutions Y77S and E167D in SEQ ID NO: 1.
6. The method of claim 1, wherein the linker peptide comprises one or more amino acid residues having a functional group suitable for conjugation to a heterologous molecule, and wherein the amino acid residue having the functional group is a cysteine, lysine, histidine, or a non-natural amino acid residue.
7. The method of claim 1, further comprising one or more amino acid residues having a functional group suitable for conjugation to a second heterologous molecule in the linker peptide, and wherein the amino acid residue having the functional group is a cysteine, lysine, histidine, or a non-natural amino acid residue.
8. The method of claim 1, wherein the binding region comprises one or more amino acid residues having a functional group suitable for conjugation to a heterologous molecule, and wherein the amino acid residue having the functional group is a cysteine, lysine, histidine, or a non-natural amino acid residue.
9. The method of claim 8, wherein the binding region comprises a single-chain variable fragment comprising a heavy chain variable domain (V.sub.H) and a light chain variable domain (V.sub.L), and wherein a linker between the V.sub.H and the V.sub.L comprises the one or more amino acid residues having the functional group.
10. The method of claim 1, wherein the Shiga toxin effector polypeptide comprises the amino acid substitution S8C in SEQ ID NO: 1.
11. The method of claim 7, wherein the amino acid residue having the functional group is a lysine, a selenocysteine, or a pyrroline-carboxy-lysine.
12. The method of claim 7, wherein the amino acid residue having the functional group comprises a reactive chemical group.
13. The method of claim 1, wherein the Shiga toxin effector polypeptide comprises SEQ ID NO: 86 or SEQ ID NO: 106.
14. The method of claim 1, wherein the linker peptide comprises the amino acid sequence of any one of SEQ ID NOs: 757-761 and 769-772.
15. The method of claim 1, wherein the binding region comprises a: single-domain antibody fragment, single-chain variable fragment, antibody variable fragment, complementary determining region 3 fragment, constrained FR3-CDR3-FR4 polypeptide, Fd fragment, antigen-binding fragment, Armadillo repeat polypeptide, fibronectin-derived 10.sup.th fibronectin type III domain, tenascin type III domain, ankyrin repeat motif domain, low-density-lipoprotein-receptor-derived A-domain, lipocalin, Kunitz domain, Protein-A-derived Z domain, gamma-B crystallin-derived domain, ubiquitin-derived domain, Sac7d-derived polypeptide, Fyn-derived SH2 domain, miniprotein, or C-type lectin-like domain scaffold.
16. The method of claim 1, wherein the binding region comprises a single-chain variable fragment or single-domain antibody fragment.
17. The method of claim 1, wherein the extracellular target biomolecule is: B7-H3, BCMA, CD20, PD-L1, CD22, CD40, CD45, CD79, CD25, CD30, HER2/neu/ErbB2, EGFR, EphB2, prostate-specific membrane antigen, Cripto, endoglin/CD105, fibroblast activation protein, Lewis-Y, CD19, CD21, CS1/SLAMF7, CD33, CD52, CD74, EpCAM, gpA33, mucin, TAG-72, carbonic anhydrase IX, folate binding protein, ganglioside GD2, ganglioside GD3, ganglioside GM2, ganglioside Lewis-Y2, VEGFR, Alpha Vbeta3, Alpha5beta1, ErbB1/EGFR, Erb3, c-MET, IGF1R, EphA3, LMP1, TRAIL-R1, TRAIL-R2, RANKL, tenascin, CD64, mesothelin, BRCA1, MART-1/MelanA, gp100, tyrosinase, TRP-1, TRP-2, MAGE-1, MAGE-3, GAGE-1/2, BAGE, RAGE, NY-ESO-1, CDK-4, beta-catenin, MUM-1, caspase-8, KIAA0205, HPVE6, SART-1, PRAME, carcinoembryonic antigen, prostate specific antigen, prostate stem cell antigen, human aspartyl (asparaginyl) beta-hydroxylase, EphA2, HER3/ErbB-3, MUC1, tyrosinase associated antigen, human tyrosinase-related protein 1, HPV-E7, Epstein-Barr virus antigen, Bcr-Abl, alpha-fetoprotein antigen, 17-A1, bladder tumor antigen, CD38, CD15, CD23, CD53, CD88, CD129, CD183, CD191, CD193, CD244, CD294, CD305, C3AR, FceRIa, galectin-9, mrp-14, siglec-8, siglec-10, CD49d, CD13, CD44, CD54, CD63, CD69, CD123, TLR4, IgE, CD107a, CD203c, CD14, CD68, CD80, CD86, CD115, F4/80, ILT-3, galectin-3, CD11a-c, GITRL, MHC class II, CD284/TLR4, CD107/Mac3, CD195/CCR5, HLA-DR, CD16/32, or CD282/TLR2.
18. The method of claim 1, wherein the cell-targeting molecule is covalently linked via the one or more cysteines to the heterologous molecule.
19. The method of claim 18, wherein the heterologous molecule is an antibiotic, antigen, cytotoxic agent, radionuclide, solubility-altering agent, pharmacokinetic-altering agent, immunogenicity-altering agent, pharmacodynamics-altering agent, detection-promoting agent, dye, T-cell epitope, fluorophore, immunogen, enzyme, zymoxin, lipid, polymer, polyethylene glycol, serum albumin binding agent, small molecule chemotherapeutic agent, prodrug, peptide, protein, nucleic acid, or protein-nucleic acid complex.
20. The method of claim 18, wherein the heterologous molecule is an auristatin, a pyrrolobenzodiazepine or a pyrrolobenzodiazepine dimer.
21. The method of claim 1, wherein the cancer is selected from the group consisting of: bone cancer, breast cancer, central/peripheral nervous system cancer, gastrointestinal cancer, germ cell cancer, glandular cancer, head-neck cancer, hematological cancers, kidney-urinary tract cancer, liver cancer, lung/pleura cancer, prostate cancer, sarcoma, skin cancer, and uterine cancer.
Description
BRIEF DESCRIPTION OF THE FIGURES
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DETAILED DESCRIPTION
[0197] The present invention is described more fully hereinafter using illustrative, non-limiting embodiments, and references to the accompanying figures. This invention may, however, be embodied in many different forms and should not be construed as to be limited to the embodiments set forth below. Rather, these embodiments are provided so that this disclosure is thorough and conveys the scope of the invention to those skilled in the art.
[0198] In order that the present invention may be more readily understood, certain terms are defined below. Additional definitions may be found within the detailed description of the invention.
[0199] As used in the specification and the appended claims, the terms a, an and the include both singular and the plural referents unless the context clearly dictates otherwise.
[0200] As used in the specification and the appended claims, the term and/or when referring to two species, A and B, means at least one of A and B. As used in the specification and the appended claims, the term and/or when referring to greater than two species, such as A, B, and C, means at least one of A, B, or C, or at least one of any combination of A, B, or C (with each species in singular or multiple possibility).
[0201] Throughout this specification, the word comprise or variations such as comprises or comprising will be understood to imply the inclusion of a stated integer (or components) or group of integers (or components), but not the exclusion of any other integer (or components) or group of integers (or components).
[0202] Throughout this specification, the term including is used to mean including but not limited to. Including and including but not limited to are used interchangeably.
[0203] The term amino acid residue or amino acid includes reference to an amino acid that is incorporated into a protein, polypeptide, or peptide. The term polypeptide includes any polymer of amino acids or amino acid residues. The term polypeptide sequence refers to a series of amino acids or amino acid residues from which a polypeptide is physically composed. A protein is a macromolecule comprising one or more polypeptides or polypeptide chains. A peptide is a small polypeptide of sizes less than about 15 to 20 amino acid residues. The term amino acid sequence refers to a series of amino acids or amino acid residues which physically comprise a peptide or polypeptide depending on the length. Unless otherwise indicated, polypeptide and protein sequences disclosed herein are written from left to right representing their order from an amino terminus to a carboxy terminus.
[0204] For purposes of the claimed invention and with regard to a Shiga toxin protein sequence or Shiga toxin derived polypeptide, the term wild-type generally refers to a naturally occurring, Shiga toxin protein sequence(s) found in a living species, such as, e.g., a pathogenic bacterium, wherein that Shiga toxin protein sequence(s) is one of the most frequently occurring variants. This is in contrast to infrequently occurring Shiga toxin protein sequences that, while still naturally occurring, are found in less than one percent of individual organisms of a given species out of individual organisms of that same species when sampling a statistically powerful number of naturally occurring individual organisms of that species which comprise at least one Shiga toxin protein variant. A clonal expansion of a natural isolate outside its natural environment (regardless of whether the isolate is an organism or molecule comprising biological sequence information) does not alter the naturally occurring requirement as long as the clonal expansion does not introduce new sequence variety not present in naturally occurring populations of that species and/or does not change the relative proportions of sequence variants to each other.
[0205] The terms amino acid, amino acid residue, amino acid sequence, or polypeptide sequence include naturally occurring amino acids (including L and D isosteriomers) and, unless otherwise limited, also include known analogs of the twenty common natural amino acids that can function in a similar manner, such as, e.g., selenocysteine, pyrrolysine, N-formylmethionine, gamma-carboxyglutamate, hydroxyprolinehypusine, pyroglutamic acid, and selenomethionine (see e.g. Nagata K et al., Bioinformatics 30: 1681-9 (2014)). The amino acids referred to herein are described by shorthand designations as follows in Table A:
TABLE-US-00001 TABLE A Amino Acid Nomenclature Name 3-letter 1-letter Alanine Ala A Arginine Arg R Asparagine Asn N Aspartic Acid or Aspartate Asp D Cysteine Cys C Glutamic Acid or Glutamate Glu E Glutamine Gln Q Glycine Gly G Histidine His H Isoleucine Ile I Leucine Leu L Lysine Lys K Methionine Met M Phenylalanine Phe F Proline Pro P Pyrroline-carboxy-lysine Pcl Selenocysteine Sec Serine Ser S Threonine Thr T Tryptophan Trp W Tyrosine Tyr Y Valine Val V
[0206] The phrase conservative substitution with regard to a protein, polypeptide, or polypeptide region refers to a change in the amino acid composition of the polypeptide that does not substantially alter the function and structure of the overall protein, polypeptide, or polypeptide region (see Creighton, Proteins: Structures and Molecular Properties (W. H. Freeman and Company, New York (2nd ed., 1992)).
[0207] As used herein, the terms expressed, expressing, or expresses, and grammatical variants thereof, refer to translation of a polynucleotide or nucleic acid into a protein. The expressed protein may remain intracellular, become a component of the cell surface membrane or be secreted into an extracellular space.
[0208] As used herein, the phrase target-expressing cell encompasses any cell that expresses, at a cellular surface, a target biomolecule bound by a binding region of the cell-targeting molecule of the present invention.
[0209] As used herein, cells which express a significant amount of an extracellular target biomolecule at least one cellular surface are target positive cells or target+ cells and are cells physically coupled to the specified, extracellular target biomolecule.
[0210] As used herein, cells which express a significant amount of target biomolecule at least one cellular surface are target-positive cells or target+ cells and are cells physically coupled to the extracellular target biomolecule. A significant amount of target biomolecule is defined below.
[0211] As used herein, the symbol is shorthand for an immunoglobulin-type binding region capable of binding to the biomolecule following the symbol. The symbol is used to refer to the functional characteristic of an immunoglobulin-type binding region based on its ability to bind to the biomolecule following the symbol with a binding affinity described by a dissociation constant (K.sub.D) of 10.sup.5 or less.
[0212] The terms associated, associating, linked, or linking with regard to the claimed invention refers to the state of two or more components of a molecule being joined, attached, connected, or otherwise coupled to form a single molecule or the act of making two molecules associated with each other to form a single molecule by creating an association, linkage, attachment, and/or any other connection between the two molecules. For example, the term linked may refer to two or more components associated by one or more atomic interactions such that a single molecule is formed and wherein the atomic interactions may be covalent and/or non-covalent. Non-limiting examples of covalent associations between two components include peptide bonds and cysteine-cysteine disulfide bonds. Non-limiting examples of non-covalent associations between two molecular components include ionic bonds.
[0213] For purposes of the present invention, the term linked refer to two or more molecular components associated by one or more atomic interactions such that a single molecule is formed and wherein the atomic interactions includes at least one covalent bond. For purposes of the present invention, the term linking refers to the act of creating a linked molecule as described above.
[0214] For purposes of the present invention, the term fused refers to two or more proteinaceous components associated by at least one covalent bond which is a peptide bond, regardless of whether the peptide bond involves the participation of a carbon atom of a carboxyl acid group or involves another carbon atom, such as, e.g., the -carbon, -carbon, -carbon, -carbon, etc. Non-limiting examples of two proteinaceous components fused together include, e.g., an amino acid, peptide, or polypeptide fused to a polypeptide via a peptide bond such that the resulting molecule is a single, continuous polypeptide. For purposes of the present invention, the term fusing refers to the act of creating a fused molecule as described above, such as, e.g., a fusion protein generated from the recombinant fusion of genetic regions which when translated produces a single proteinaceous molecule.
[0215] The symbol :: means the proteinaceous molecules before and after the symbol are physically linked together to form a continuous polypeptide.
[0216] The symbol _ with regard to a cell-targeting molecule means the molecules before and after the symbol are covalently linked together, either directly or indirectly.
[0217] For purposes of the present invention, the term effector means providing a biological activity, such as cytotoxicity, biological signaling, enzymatic catalysis, subcellular routing, and/or intermolecular binding resulting in the recruitment of one or more factors and/or allosteric effect(s). For example, a Shiga toxin effector polypeptide provides one or more biological activities present in a Shiga toxin, Shiga toxin component, and/or fragment thereof.
[0218] As used herein, the phrase multivalent target-binding molecule refers to a target-binding molecule or plurality of target-binding molecules comprising two or more high-affinity binding regions, such as, e.g. a protein comprising two or more binding regions wherein each individual binding region has a dissociation constant of 10.sup.5 to 10.sup.12 moles per liter toward an extracellular part of a target biomolecule.
[0219] For purposes of the present invention, the phrase derived from when referring to a polypeptide or polypeptide region means that the polypeptide or polypeptide region comprises amino acid sequences originally found in a parental protein and which may now comprise certain amino acid residue additions, deletions, truncations, rearrangements, or other alterations relative to the original polypeptide or polypeptide region as long as a certain function(s) and a structure(s) of the parental molecule are substantially conserved. The skilled worker will be able to identify a parental molecule from which a polypeptide or polypeptide region was derived using techniques known in the art, e.g., protein sequence alignment software.
[0220] For purposes of the present invention, a Shiga toxin effector function is a biological activity conferred by a polypeptide region derived from a Shiga toxin A Subunit. Non-limiting examples of Shiga toxin effector functions include cellular internalization, subcellular routing, catalytic activity, and cytotoxicity. Shiga toxin catalytic activities include, for example, ribosome inactivation, protein synthesis inhibition, N-glycosidase activity, polynucleotide:adenosine glycosidase activity, RNAase activity, and DNAase activity. Shiga toxins are ribosome inactivating proteins (RIPs). RIPs can depurinate nucleic acids, polynucleosides, polynucleotides, rRNA, ssDNA, dsDNA, mRNA (and polyA), and viral nucleic acids (see e.g. Brigotti M et al., Toxicon 39: 341-8 (2001); Brigotti M et al., FASEB J 16: 365-72 (2002)). Some RIPs show antiviral activity and superoxide dismutase activity. Shiga toxin catalytic activities have been observed both in vitro and in vivo. Non-limiting examples of assays for Shiga toxin effector activity measure protein synthesis inhibitory activity, depurination activity, inhibition of cell growth, cytotoxicity, supercoiled DNA relaxation activity, and nuclease activity.
[0221] As used herein, the retention of Shiga toxin effector function refers to being capable of exhibiting a level of Shiga toxin functional activity, as measured by an appropriate quantitative assay with reproducibility, comparable to a wild-type, Shiga toxin effector polypeptide control (e.g. a Shiga toxin A1 fragment) or cell-targeting molecule comprising a wild-type Shiga toxin effector polypeptide (e.g. a Shiga toxin A1 fragment) under the same conditions. For the Shiga toxin effector function of ribosome inactivation or ribosome inhibition, retained Shiga toxin effector function is exhibiting an IC.sub.50 of 10,000 picomolar (pM) or less in an in vitro setting, such as, e.g., by using an assay known to the skilled worker and/or described herein. For the Shiga toxin effector function of cytotoxicity in a target positive cell-kill assay, retained Shiga toxin effector function is exhibiting a CD.sub.50 of 1,000 nanomolar (nM) or less, depending on the cell type and its expression of the appropriate extracellular target biomolecule, as shown, e.g., by using an assay known to the skilled worker and/or described herein.
[0222] As used herein, the retention of significant Shiga toxin effector function refers to a level of Shiga toxin functional activity, as measured by an appropriate quantitative assay with reproducibility comparable to a wild-type Shiga toxin effector polypeptide control. For in vitro ribosome inhibition, significant Shiga toxin effector function is exhibiting an IC.sub.50 of 300 pM or less depending on the source of the ribosomes (e.g. bacteria, archaea, or eukaryote (algae, fungi, plants, or animals)). This is significantly greater inhibition as compared to the approximate IC.sub.50 of 100,000 pM for the catalytically inactive SLT-1A 1-251 double mutant (Y77S/E167D). For cytotoxicity in a target-positive cell kill assay in laboratory cell culture, significant Shiga toxin effector function is exhibiting a CD.sub.50 of 100, 50, or 30 nM or less, depending on the cell line and its expression of the appropriate extracellular target biomolecule. This is significantly greater cytotoxicity to the appropriate target cell line as compared to an SLT-1A subunit alone, without a cell targeting binding region, which has a CD.sub.50 of 100-10,000 nM, depending on the cell line.
[0223] For some samples, accurate values for either IC.sub.50 or CD.sub.50 might be unobtainable due to the inability to collect the required data points for an accurate curve fit. For example, theoretically, neither an IC.sub.50 nor CD.sub.50 can be determined if greater than 50% ribosome inhibition or cell death, respectively, does not occur in a concentration series for a given sample. Data insufficient to accurately fit a curve as described in the analysis of the data from exemplary Shiga toxin effector function assays, such as, e.g., assays described in the Examples, should not be considered as representative of actual Shiga toxin effector function.
[0224] A failure to detect activity in Shiga toxin effector function may be due to improper expression, polypeptide folding, and/or polypeptide stability rather than a lack of cell entry, subcellular routing, and/or enzymatic activity. Assays for Shiga toxin effector functions may not require much cell-targeting molecule of the invention to measure significant amounts of Shiga toxin effector function activity. To the extent that an underlying cause of low or no effector function is determined empirically to relate to protein expression or stability, one of skill in the art may be able to compensate for such factors using protein chemistry and molecular engineering techniques known in the art, such that a Shiga toxin functional effector activity may be restored and measured. As examples, improper cell-based expression may be compensated for by using different expression control sequences; improper polypeptide folding and/or stability may benefit from stabilizing terminal sequences, or compensatory mutations in non-effector regions which stabilize the three-dimensional structure of the protein, etc. When new assays for individual Shiga toxin functions become available, Shiga toxin effector regions or polypeptides may be analyzed for any level of those Shiga toxin effector functions, such as for being within a certain-fold activity of a wild-type Shiga toxin effector polypeptide. Examples of meaningful activity differences are, e.g., Shiga toxin effector regions that have 1000-fold or 100-fold or less the activity of a wild-type Shiga toxin effector polypeptide; or that have 3-fold to 30-fold or more activity compared to a functional knock-down or knockout Shiga toxin effector polypeptide.
[0225] Certain Shiga toxin effector functions are not easily measurable, e.g. subcellular routing functions. Currently there is no routine, quantitative assay to distinguish whether the failure of a Shiga toxin effector polypeptide to be cytotoxic is due to improper subcellular routing, but at a time when tests are available, Shiga toxin effector polypeptides may be analyzed for any significant level of subcellular routing as compared to the appropriate wild-type Shiga toxin effector region.
[0226] It should be noted that even if the cytotoxicity of a Shiga toxin effector polypeptide is reduced relative to wild-type, in practice, applications using attenuated, Shiga toxin effector polypeptides may be equally or more effective than those using wild-type, Shiga toxin effector polypeptides because the highest potency variants might exhibit undesirable effects which are minimized or reduced in reduced-potency variants. Wild-type Shiga toxin effector polypeptides are very potent, being able to kill with only one molecule reaching the cytosol or perhaps 40 molecules being internalized (Tam P, Lingwood C, Microbiology 153: 2700-10 (2007)). Shiga toxin effector polypeptides with even considerably reduced Shiga toxin effector functions, such as, e.g., subcellular routing or cytotoxicity, as compared to wild-type Shiga toxin effector polypeptides may still be potent enough for practical applications involving targeted cell killing and/or detection of certain subcellular compartments of specific cell types. And such effector polypeptides may also be useful for delivering cargos (e.g. additional exogenous material) to certain intracellular locations or subcellular compartments.
[0227] The term selective cytotoxicity with regard to the cytotoxic activity of a cytotoxic, cell-targeting molecule refers to the relative levels of cytotoxicity between a targeted cell population and a non-targeted bystander cell population, which can be expressed as a ratio of the half-maximal cytotoxic concentration (CD.sub.50) for a targeted cell type over the CD.sub.50 for an untargeted cell type to show the preferentiality of cell killing of the targeted cell type as a metric for selectivity.
[0228] As used in the specification and the claims herein, the phrase physiological temperature appropriate for the cell refers to temperatures known in the art and/or identifiable by the skilled worker which fall within a range suitable for healthy growth, propagation, and/or function of that particular cell or cell type; corresponding to the core temperature of the species from which the cell is derived; and/or corresponding to a healthy, living organism comprising the cell. For example, temperatures around 37 C. are appropriate for many mammalian cells depending on the species.
[0229] For purposes of the present invention, the phrase internalization of a molecular complex comprising the cell-targeting molecule bound to target biomolecule means the cellular internalization of the cell-targeting molecule is target-mediated in that the internalization begins with the cell-targeting molecule and cell-surface target biomolecule forming a complex at an extracellular position and ends with both the cell-targeting molecule and target biomolecule(s) entering the cell prior to dissociation of the cell-targeting molecule from the target biomolecule(s) to which the cell-targeting molecule has bound.
[0230] For purposes of the present invention, the phrase target biomolecule natively present on the surface of a cell means a cell expresses the target biomolecule using its own internal machinery and localizes the target biomolecule to a cellular surface using its own internal machinery such that the target biomolecule is physically coupled to said cell and at least a part of the target biomolecule is accessible from an extracellular space, i.e. on the surface of a cell.
[0231] For purposes of the present invention, the phrase cell-targeting molecule altering agent refers to any of a number of different types of atoms or molecules known to the skilled worker and/or described herein which may be conjugated to a molecule of the invention in order to alter one or more properties of the molecule of the invention.
[0232] For the purposes of certain embodiments of the present invention, cellular internalization is considered rapid if the time for internalization to occur due to the binding of the cell-targeting molecule of the present invention is reduced as compared to the time for internalization of a prior art reference molecule at the same percent target biomolecule occupancy as determined by the same assay using the same cell type at the same temperature.
[0233] As used in the specification and the claims herein, the phrase rapid cellular internalization refers to the ability of a cell-targeting molecule of the present invention to decrease the time on average for cellular internalization of an extracellular target antigen or cell surface localized target biomolecule as compared to the time on average required for cellular internalization of an extracellular target antigen or cell surface localized target biomolecule, as measured by any one of a number of cell internalization assays known in the art or described herein.
[0234] As used in the specification and the claims herein, the phrase rapid internalization includes internalization which may be assayed as compared to a basal target biomolecule internalization rate and/or molecular binding induced internalization rate for target biomolecule after administration of an immunoglobulin-type binding molecule (e.g. a monoclonal antibody) known in the art to bind an extracellular part of target biomolecule. The scope of the phrase rapid cellular internalization is intended to encompass internalization rates, on average, faster than those observed when testing a target-specific antibody or immunoglobulin-derived protein molecule with an Fc region. In general, an internalization rate constant may be defined as the time after administration of a target-specific binding molecule of interest to target-positive cells at which 50% of cell surface target antigens, target biomolecules, and/or the target-specific binding molecule is internalized at a given administered concentration, mass, molarity, or target biomolecule occupancy-adjusted concentration, to a particular cell type, and at a particular temperature. Cell-surface target biomolecule internalization, whether basally or in response to administration of a target-binding molecule, may be assayed by various methods known to the skilled worker.
[0235] For the purposes of certain embodiments of the present invention, cellular internalization is considered rapid if the time for internalization to occur due to the binding of the cell-targeting molecule of the present invention is reduced as compared to the time for internalization of the target biomolecule with the binding of a well-characterized antibody recognizing an extracellular target biomolecule antigen. The term rapid as used throughout the present description is intended to indicate that a cell-targeting molecule of the present invention enters one or more target-expressing and/or target-positive cells in less than six hours. In certain embodiments, rapid can be as quickly as less than about thirty minutes, but can also encompass a range of from about 1 hour to about 2 hours, to about 3 hours, to about 4 hours, to about 5 hours; a range of about 2 hours to about 3 hours, to about 4 hours, to about 5 hours; a range of about 3 hours to about 4 hours, to about 5 hours; and a range of about 4 hours to about 5 hours.
[0236] For purposes of the present invention, the phrase one or more non-covalent linkages, with regard to a molecule comprising two or more components linked together, includes the types of linkages connecting the components that in certain molecules may be observed as being eliminated (i.e., no longer connecting two or more components) when changing the molecule from native protein-folding conditions to protein-denaturing conditions. For example, when using techniques known in the art and/or described herein, such as, e.g., electrophoretic and/or chromatographic assays, for assaying the sizes of proteinaceous molecules, a multi-component molecule that appears as a single-sized species under native protein-folding conditions (e.g. pH-buffered environments intended to be similar to the lumen of the endoplasmic reticulum of a eukaryotic cell or to an extracellular environment within an organism), can also be observed as being composed of two or more smaller-sized, proteinaceous molecules under denaturing conditions and/or after being subjected to a denaturing condition. Protein-denaturing conditions are known to the skilled worker and include conditions markedly different from native protein-folding conditions, such as, e.g., environments with a high temperature (e.g., greater than 50 degrees Celsius) and/or those characterized by the presence of chemical denaturants and/or detergents, such as, e.g., 1-10% sodium dodecyl sulfate, polysorbates, Triton X-100, sarkosyl, and other detergents whether ionic, non-ionic, zwitterionic, and/or chaotropic.
[0237] As used herein, the term monomeric with regard to describing a protein and/or proteinaceous molecule refers to a molecule comprising only one polypeptide component consisting of a single, continuous polypeptide, regardless of its secondary or tertiary structure, which may be synthesized by a ribosome from a single polynucleotide template, including a continuous linear polypeptide which later forms a cyclic structure. In contrast, a multimeric molecule comprises two or more polypeptides (e.g. subunits) which together do not form a single, continuous polypeptide that may be synthesized by a ribosome from a single polynucleotide template.
[0238] As used herein, the term multimeric with regard to describing a protein and/or proteinaceous molecule refers to a molecule that comprises two or more, individual, polypeptide components associated together and/or linked together, such as, e.g., a molecule consisting of two components each of which is its own continuous polypeptide. For example, the association or linkage between components of a molecule may include 1) one or more non-covalent interactions; 2) one or more post-translational, covalent interactions; 3) one or more, covalent chemical conjugations; and/or 4) one or more covalent interactions resulting in a single molecule comprising a non-linear polypeptide, such as, e.g., a branched or cyclic polypeptide structure, resulting from the arrangement of the two or more polypeptide components. A molecule comprising two, discontinuous polypeptides as a result of the proteolytic cleavage of one or more peptide bonds in a single, continuous polypeptide synthesized by a ribosome from a single polynucleotide templates is multimeric and not monomeric.
[0239] As used herein, the terms disrupted, disruption, or disrupting, and grammatical variants thereof, with regard to a polypeptide region or feature within a polypeptide refers to an alteration of at least one amino acid within the region or composing the disrupted feature. Amino acid alterations include various mutations, such as, e.g., a deletion, inversion, insertion, or substitution which alter the amino acid sequence of the polypeptide. Amino acid alterations also include chemical changes, such as, e.g., the alteration one or more atoms in an amino acid functional group or the addition of one or more atoms to an amino acid functional group.
[0240] As used herein, de-immunized means reduced antigenic and/or immunogenic potential after administration to a chordate as compared to a reference molecule, such as, e.g., a wild-type peptide region, polypeptide region, or polypeptide. This includes a reduction in overall antigenic and/or immunogenic potential despite the introduction of one or more, de novo, antigenic and/or immunogenic epitopes as compared to a reference molecule. For certain embodiments, de-immunized means a molecule exhibited reduced antigenicity and/or immunogenicity after administration to a mammal as compared to a parental molecule from which it was derived, such as, e.g., a wild-type Shiga toxin A1 fragment. In certain embodiments, the de-immunized, Shiga toxin effector polypeptide of the present invention is capable of exhibiting a relative antigenicity compared to a reference molecule which is reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or greater than the antigenicity of the reference molecule under the same conditions measured by the same assay, such as, e.g., an assay known to the skilled worker and/or described herein like a quantitative ELISA or Western blot analysis. In certain embodiments, the de-immunized, Shiga toxin effector polypeptide of the present invention is capable of exhibiting a relative immunogenicity compared to a reference molecule which is reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 99%, or greater than the immunogenicity of the reference molecule under the same conditions measured by the same assay, such as, e.g., an assay known to the skilled worker and/or described herein like a quantitative measurement of anti-molecule antibodies produced in a mammal(s) after receiving parenteral administration of the molecule at a given time-point. The relative immunogenicities of exemplary cell-targeting molecules may be determined using an assay for in vivo antibody responses to the cell-targeting molecules after repeat, parenteral administrations over periods of many.
[0241] For purposes of the present invention, the terms terminus, amino-terminus, or carboxy-terminus with regard to a cell-targeting molecule refers generally to the last amino acid residue of a polypeptide chain of the cell-targeting molecule (e.g., a single, continuous polypeptide chain). A cell-targeting molecule may comprise more than one polypeptides or proteins, and, thus, a cell-targeting molecule of the present invention may comprise multiple amino-terminals and carboxy-terminals. For example, the amino-terminus of a cell-targeting molecule may be defined by the first amino acid residue of a polypeptide chain representing the amino-terminal end of the polypeptide, which is generally characterized by a starting, amino acid residue which does not have a peptide bond with any amino acid residue involving the primary amino group of the starting amino acid residue or involving the equivalent nitrogen for starting amino acid residues which are members of the class of N-alkylated alpha amino acid residues. Similarly, the carboxy-terminus of a cell-targeting molecule may be defined by the last amino acid residue of a polypeptide chain representing the carboxyl-terminal end of the polypeptide, which is generally characterized by a final, amino acid residue which does not have any amino acid residue linked by a peptide bond to the alpha-carbon of its primary carboxyl group.
[0242] For purposes of the present invention, the terms terminus, amino-terminus, or carboxy-terminus with regard to a polypeptide region refers to the regional boundaries of that region, regardless of whether additional amino acid residues are linked by peptide bonds outside of that region. In other words, the terminals of the polypeptide region regardless of whether that region is fused to other peptides or polypeptides. For example, a fusion protein comprising two proteinaceous regions, e.g., a binding region comprising a peptide or polypeptide and a Shiga toxin effector polypeptide, may have a Shiga toxin effector polypeptide region with a carboxy-terminus ending at amino acid residue 251 of the Shiga toxin effector polypeptide region despite a peptide bond involving residue 251 to an amino acid residue at position 252 representing the beginning of another proteinaceous region, e.g., the binding region. In this example, the carboxy-terminus of the Shiga toxin effector polypeptide region refers to residue 251, which is not a terminus of the fusion protein but rather represents an internal, regional boundary. Thus, for polypeptide regions, the terms terminus, amino-terminus, and carboxy-terminus are used to refer to the boundaries of polypeptide regions, whether the boundary is a physically terminus or an internal, position embedded within a larger polypeptide chain.
[0243] For purposes of the present invention, the phrase furin-cleavage resistant means a molecule or specific polypeptide region thereof exhibits reproducibly less furin cleavage than (i) the carboxy-terminus of a Shiga toxin A1 fragment in a wild-type Shiga toxin A Subunit or (ii) the carboxy-terminus of the Shiga toxin A1 fragment derived region of construct wherein the naturally occurring furin-cleavage site natively positioned at the junction between the A1 and A2 fragments is not disrupted; as assayed by any available means to the skilled worker, including by using a method described herein.
[0244] For purposes of the present invention, the phrase disrupted furin-cleavage motif refers to (i) a specific furin-cleavage motif as described herein in Section I-B and (ii) which comprises a mutation and/or truncation that can confer a molecule with a reduction in furin-cleavage as compared to a reference molecule, such as, e.g., a reduction in furin-cleavage reproducibly observed to be 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99%, or less (including 100% for no cleavage) than the furin-cleavage of a reference molecule observed in the same assay under the same conditions. The percentage of furin-cleavage as compared to a reference molecule can be expressed as a ratio of cleaved:uncleaved material of the molecule of interest divided by the cleaved:uncleaved material of the reference molecule (see e.g. WO 2015/191764). Non-limiting examples of suitable reference molecules include certain molecules comprising a wild-type Shiga toxin furin-cleavage motif and/or furin-cleavage site are described in WO 2015/191764 and WO 2016/196344.
Introduction
[0245] The present invention provides Shiga toxin effector polypeptides and cell-targeting molecules comprising specific attachment sites for conjugating other molecules. A unique amino acid residue with a unique functional group that is solvent accessible and/or one or more position-ectopic residues in a Shiga toxin effector polypeptide or cell-targeting molecule scaffold provides an attachment point for the site-specific linking of various atoms and molecules. The atoms or molecules may function as (1) cargos designed for intracellular delivery, including for controlled liberation once inside a target cell, and/or (2) agents having extracellular function(s), such as, e.g., biologically inert moieties which prolong half-life in a vertebrate, mask immunogenic portions of the scaffold, and/or block proteolytic cleavage. The Shiga toxin effector polypeptides and cell-targeting molecules of the present invention may be conjugated to a variety of atoms and molecules in a controlled and convenient manner using routine methods in order to obtain homogenous products.
[0246] The present invention also provides Shiga toxin effector polypeptides and cell-targeting molecules conjugated to other molecules, such as, e.g., a molecular cargo for intracellular delivery or a cell-targeting molecule altering agent. Certain cell-targeting molecules of the present invention, and compositions thereof, may be used to selectively deliver conjugated cargo(s) to a target-expressing cell type(s) in the presence of one or more other cell types based on its cell-targeting and cellular internalization activity(ies), such as, e.g., a cargo having a desired, intracellular function. In addition, certain cell-targeting molecules of the present invention, and compositions thereof, may be used to selectively kill a target-expressing cell in the presence of one or more other cell types based on its cell-targeting activity and cellular internalization activity(ies), such as, e.g., by delivering into the interior of the targeted, target-expressing cell a component of the cell-targeting molecule which is cytotoxic at an intracellular location. The conjugated atoms, molecules, cargos, and/or cell-targeting molecule altering agents contemplated as suitable for use in the present invention include linkers, cell-targeting moieties, antibiotics, peptides, nucleic acids, proteins, protein-nucleic acid complexes, cytotoxic agents, solubility-altering agents, pharmacokinetic-altering agents, immunogenicity-altering agents, and pharmacodynamics-altering agents.
[0247] The molecules of the present invention, and compositions thereof, have uses, e.g., for the selective delivery of cargos to target-expressing cells and as therapeutics for the treatment of a variety of diseases, disorders, and conditions.
I. General Structures of the Shiga Toxin Effector Polypeptides and Cell-Targeting Molecules of the Present Invention
[0248] Certain embodiments of the present invention are Shiga toxin A Subunit effector polypeptides, such as, e.g., (1) a Shiga toxin effector polypeptide conjugated to a heterologous molecule; and (2) a Shiga toxin effector polypeptide comprising one or more ectopic amino acid residues relative to wild-type, Shiga toxin polypeptides. The present invention provides cell-targeting molecules comprising (1) a cell-targeting moiety (e.g. a cell-targeting agent and/or binding region) and (2) a toxin effector polypeptide region. Certain further embodiments are cell-targeting molecules comprising a Shiga toxin effector polypeptide of the present invention. In addition, the present invention provides cell-targeting molecules lacking any Shiga toxin effector polypeptide but comprising a linker or binding region (e.g. an immunoglobulin-type polypeptide) comprising functional group(s) for site specific attachment of other molecules. All of the molecules of the present invention may be optionally conjugated to another molecule, such as, e.g., a cargo for cell-targeted delivery, a cell-targeting molecule altering agent, and/or an additional exogenous material.
A. Shiga Toxin Effector A Subunit Polypeptides of the Present Invention
[0249] A Shiga toxin effector polypeptide of the present invention is a polypeptide derived from a Shiga toxin A Subunit of at least one member of the Shiga toxin family wherein the Shiga toxin effector region is capable of exhibiting at least one Shiga toxin function. Shiga toxin functions include, e.g., promoting cell entry, deforming lipid membranes, stimulating clathrin-mediated endocytosis, directing retrograde transport, directing subcellular routing, avoiding intracellular degradation, catalytically inactivating ribosomes, effectuating cytotoxicity, and effectuating cytostatic effects.
[0250] There are numerous Shiga toxin effector polypeptides known to the skilled worker (see e.g., Cheung M et al., Mol Cancer 9: 28 (2010); WO 2014/164693; WO 2015/113005; WO 2015/113007; US20150259428; WO 2015/191764; US20160177284; WO 2016/126950) that are suitable for use in the present invention or to use as parental polypeptides to be modified into a Shiga toxin effector polypeptide of the present invention using techniques known the art.
[0251] Shiga toxin effector polypeptides of the present invention comprise or consist essentially of a polypeptide derived from a Shiga toxin A Subunit dissociated from any form of its native Shiga toxin B Subunit. In addition, the cell-targeting molecules of the present invention need not comprise any polypeptide comprising or consisting essentially of a functional binding domain of a native Shiga toxin B subunit. Rather, the Shiga toxin effector polypeptides of the present invention may be functionally associated with heterologous binding regions to effectuate cell targeting.
[0252] In certain embodiments, a Shiga toxin effector polypeptide of the present invention may comprise or consist essentially of a full-length Shiga toxin A Subunit (e.g. SLT-1A (SEQ ID NO:1), StxA (SEQ ID NO:2), or SLT-2A (SEQ ID NO:3)), noting that naturally occurring Shiga toxin A Subunits may comprise precursor forms containing signal sequences of about 22 amino acids at their amino-terminals which are removed to produce mature Shiga toxin A Subunits and are recognizable to the skilled worker. In other embodiments, the Shiga toxin effector polypeptide of the invention comprises or consists essentially of a truncated Shiga toxin A Subunit which is shorter than a full-length Shiga toxin A Subunit, such as, e.g., a truncation known in the art (see e.g., WO 2014/164693; WO 2015/113005; WO 2015/113007; WO 2015/138452; WO 2015/191764; US20160177284; WO 2016/126950).
[0253] Although derived from a wild-type Shiga toxin A Subunit polypeptide, for certain embodiments of the molecules of the present invention, the Shiga toxin effector polypeptide differs from a naturally occurring Shiga toxin A Subunit by up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 or more amino acid residues (but by no more than that which retains at least 85%, 90%, 95%, 99%, or more amino acid sequence identity).
[0254] In certain embodiments, the Shiga toxin effector polypeptide of the present invention comprises only one lysine residue. The cytotoxic activity of the SLT-1 holotoxin was not affected by the removal of all lysine residues from the A Subunit (McCluskey, A J, Shiga-like Toxin 1: Molecular Mechanism of Toxicity and Discovery of Inhibitors, thesis, University of Toronto, (2012), Appendix B). Moreover, the removal of both amino-terminal lysines of the A Subunit of SLT-1 did not affect its cytotoxicity (McCluskey, AJ, Shiga-like Toxin 1: Molecular Mechanism of Toxicity and Discovery of Inhibitors, thesis, University of Toronto, (2012), Appendix B). Thus, all but one of the naturally occurring lysine residues in SLT-1A, StxA, and/or SLT-2A may be removed by amino acid residue substitution thereby leaving only one natively occurring lysine residue in a Shiga toxin effector polypeptide. Alternatively, all of the naturally occurring lysine residues in SLT-1A, StxA, and/or SLT-2A may be removed from a Shiga toxin effector polypeptide by amino acid residue substitution and an ectopic lysine residue may be engineered into a suitable position for site-specific conjugation and retention of one or more Shiga toxin functions.
B. Cell-Targeting Molecules of the Present Invention
[0255] The cell-targeting molecules of the present invention all comprise a cell-targeting agent or moiety, such as, e.g., a binding region described herein. Cell-targeting agents or moieties of the cell-targeting molecules of the present invention comprise molecular structures, that when linked to a polypeptide of the present invention, are each capable of bringing the cell-targeting molecule within close proximity to specific cells based on molecular interactions on the surfaces of those specific cells. Cell-targeting moieties include ligand and polypeptides which bind to cell-surface targets.
[0256] One type of cell-targeting moiety is a proteinaceous binding region. Binding regions of the cell-targeted molecules of the present invention comprise one or more polypeptides capable of selectively and specifically binding an extracellular target biomolecule. Binding regions may comprise one or more various polypeptide moieties, such as ligands whether synthetic or naturally occurring ligands and derivatives thereof, immunoglobulin derived domains, synthetically engineered scaffolds as alternatives to immunoglobulin domains, and the like. The use of proteinaceous binding regions in cell-targeting molecules of the invention allows for the creation of cell-targeting molecules which are single-chain, cell-targeting proteins.
[0257] Certain embodiments of the cell-targeting molecules of the present invention comprise a cell-targeting binding region capable of specifically binding to an extracellular part of a target biomolecule physically coupled to a cell. The binding region of a cell-targeting molecule of the present invention comprises a peptide or polypeptide region capable of binding specifically to a target biomolecule. In certain embodiments, the binding region of a cell-targeting molecule of the invention comprises one or more polypeptides capable of selectively and specifically binding an extracellular target biomolecule. Binding region may comprise one or more various peptidic or polypeptide moieties, such as randomly generated peptide sequences, naturally occurring ligands or derivatives thereof, immunoglobulin derived domains, synthetically engineered scaffolds as alternatives to immunoglobulin domains, and the like.
[0258] There are numerous binding regions known in the art that are useful for targeting polypeptides to specific cell-types via their binding characteristics, such as ligands, monoclonal antibodies, engineered antibody derivatives, and engineered alternatives to antibodies (see e.g., Cheung M et al., Mol Cancer 9: 28 (2010); WO 2014/164680; WO 2014/164693; WO 2015/113005; WO 2015/113007; WO 2015/138435; WO 2015/138452; US20150259428; WO 2015/191764; US20160177284; WO 2016/126950).
[0259] According to one specific, but non-limiting aspect, the binding region of the molecule of the invention comprises a naturally occurring ligand or derivative thereof that retains binding functionality to an extracellular target biomolecule, commonly a cell surface receptor. For example, various cytokines, growth factors, and hormones known in the art may be used to target the cell-targeting molecule to the cell-surface of specific cell types expressing a cognate cytokine receptor, growth factor receptor, or hormone receptor.
[0260] According to certain other embodiments, the binding region comprises a synthetic ligand capable of binding an extracellular target biomolecule.
[0261] According to one specific, but non-limiting aspect, the binding region may comprise an immunoglobulin-type binding region. The term immunoglobulin-type binding region as used herein refers to a polypeptide region capable of binding one or more target biomolecules, such as an antigen or epitope. Binding regions may be functionally defined by their ability to bind to target molecules. Immunoglobulin-type binding regions are commonly derived from antibody or antibody-like structures; however, alternative scaffolds from other sources are contemplated within the scope of the term.
[0262] Immunoglobulin (Ig) proteins have a structural domain known as an Ig domain. Ig domains range in length from about 70-110 amino acid residues and possess a characteristic Ig-fold, in which typically 7 to 9 antiparallel beta strands arrange into two beta sheets which form a sandwich-like structure. The Ig fold is stabilized by hydrophobic amino acid interactions on inner surfaces of the sandwich and highly conserved disulfide bonds between cysteine residues in the strands. Ig domains may be variable (IgV or V-set), constant (IgC or C-set) or intermediate (IgI or I-set). Some Ig domains may be associated with a complementarity determining region or complementary determining region (CDR) which is important for the specificity of antibodies binding to their epitopes. Ig-like domains are also found in non-immunoglobulin proteins and are classified on that basis as members of the Ig superfamily of proteins. The HUGO Gene Nomenclature Committee (HGNC) provides a list of members of the Ig-like domain containing family.
[0263] As used herein, the term heavy chain variable (V.sub.H) domain or light chain variable (V.sub.L) domain respectively refer to any antibody V.sub.H or V.sub.L domain (e.g. a human V.sub.H or V.sub.L domain) as well as any derivative thereof retaining at least qualitative antigen binding ability of the corresponding native antibody (e.g. a humanized V.sub.H or V.sub.L domain derived from a native murine V.sub.H or V.sub.L domain). A V.sub.H or V.sub.L domain consists of a framework region interrupted by the three CDRs or ABRs. The framework regions serve to align the CDRs for specific binding to an epitope of an antigen. From amino-terminus to carboxyl-terminus, both V.sub.H and V.sub.L domains comprise the following framework (FR) and CDR regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. For camelid V.sub.HH fragments, IgNARs of cartilaginous fish, V.sub.NAR fragments, and derivatives thereof, there is a single heavy chain variable domain comprising the same basic arrangement: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.
[0264] An immunoglobulin-type binding region may be a polypeptide sequence of an antibody or antigen-binding fragment thereof wherein the amino acid sequence has been varied from that of a native antibody or an Ig-like domain of a non-immunoglobulin protein, for example by molecular engineering or selection by library screening. Because of the relevance of recombinant DNA techniques and in vitro library screening in the generation of immunoglobulin-type binding regions, antibodies can be redesigned to obtain desired characteristics, such as smaller size, cell entry, or other therapeutic improvements. The possible variations are many and may range from the changing of just one amino acid to the complete redesign of, for example, a variable region. Typically, changes in the variable region will be made in order to improve the antigen-binding characteristics, improve variable region stability, or reduce the potential for immunogenic responses.
[0265] There are numerous immunoglobulin-type binding regions contemplated as components of the molecules of the present invention, such as, e.g., the cell-targeting molecules of the present invention. An immunoglobulin binding region generally comprises one or more CDRs. In certain embodiments, the immunoglobulin-type binding region is derived from an immunoglobulin binding region, such as an antibody paratope capable of binding an extracellular target biomolecule. In certain other embodiments, the immunoglobulin-type binding region comprises an engineered polypeptide not derived from any immunoglobulin domain but which functions like an immunoglobulin binding region by providing high-affinity binding to an extracellular target biomolecule. This engineered polypeptide may optionally include polypeptide scaffolds comprising or consisting essentially of complementary determining regions from immunoglobulins as described herein.
[0266] There are also numerous binding regions in the prior art that are useful for targeting polypeptides to specific cell-types via their high-affinity binding characteristics. In certain embodiments, the binding region of the present proteins is selected from the group which includes single-domain antibody domains (sdAbs), nanobodies, heavy-chain antibody domains derived from camelids (V.sub.HH fragments), bivalent nanobodies, heavy-chain antibody domains derived from cartilaginous fishes, immunoglobulin new antigen receptors (IgNARs), V.sub.NAR fragments, single-chain variable (scFv) fragments, autonomous V.sub.H domains, single-domain antibody domains (sdAbs), heavy-chain antibody domains derived from camelids (V.sub.HH fragments or V.sub.H domain fragments), heavy-chain antibody domains derived from camelid V.sub.HH fragments or V.sub.H domain fragments, heavy-chain antibody domains derived from cartilaginous fishes, immunoglobulin new antigen receptors (IgNARs), V.sub.NAR fragments, single-chain variable (scFv) fragments, nanobodies, Fd fragments consisting of the heavy chain and CHI domains, single chain Fv-C.sub.H3 minibodies, dimeric C.sub.H2 domain fragments (C.sub.H2D), Fc antigen binding domains (Fcabs), isolated complementary determining region 3 (CDR3) fragments, constrained framework region 3, CDR3, framework region 4 (FR3-CDR3-FR4) polypeptides, small modular immunopharmaceutical (SMIP) domains, multimerizing V.sub.HH fragments, scFv-Fc fusions, multimerizing scFv fragments (diabodies, triabodies, tetrabodies), disulfide stabilized antibody variable (Fv) fragments, disulfide stabilized antigen-binding (Fab) fragments consisting of the V.sub.L, V.sub.H, CL and C.sub.H1 domains, bivalent nanobodies, bivalent minibodies, bivalent F(ab).sub.2 fragments (Fab dimers), bispecific tandem V.sub.HH fragments, bispecific tandem scFv fragments, bispecific nanobodies, bispecific minibodies, and any genetically manipulated counterparts of the foregoing that retain its paratope and binding function (see e.g. Ward E et al., Nature 341: 544-6 (1989); Davies J, Riechmann L, Biotechnology (NY) 13: 475-9 (1995); Reiter Y et al., Mol Biol 290: 685-98 (1999); Riechmann L, Muyldermans S, J Immunol Methods 231: 25-38 (1999); Tanha J et al., J Immunol Methods 263: 97-109 (2002); Vranken W et. al., Biochemistry 41: 8570-9 (2002); Dottorini T et al., Biochemistry 43: 622-8 (2004); Jespers L et al., J Mol Biol 337: 893-903 (2004); Jespers L et al., Nat Biotechnol 22: 1161-5 (2004); To R et al., J Biol Chem 280: 41395-403 (2005); Spinelli S et al., FEBS Lett 564: 35-40 (2004); Saerens D et al., Curr Opin Pharmacol 8: 600-8 (2008); Dimitrov D, MAbs 1: 26-8 (2009); Baral T et al., PLoS One 7: e30149 (2012); Ahmad Z et al., Clin Dev Immunol 2012: 980250 (2012); Weiner L, Cell 148: 1081-4 (2012); Richard G et al., PLoS One 8: e69495 (2013)). There are a variety of binding regions comprising polypeptides derived from the constant regions of immunoglobulins, such as, e.g., engineered dimeric Fc domains, monomeric Fcs (mFcs), V.sub.HH-Fc fusions, scFv-Fc fusions, C.sub.H2 domains, monomeric C.sub.H3s domains (mC.sub.H3s), synthetically reprogrammed immunoglobulin domains, and/or hybrid fusions of immunoglobulin domains with ligands (Hofer T et al., Proc Natl Acad Sci USA 105: 12451-6 (2008); Xiao J et al., J Am Chem Soc 131: 13616-13618 (2009); Xiao X et al., Biochem Biophys Res Commun 387: 387-92 (2009); Wozniak-Knopp G et al., Protein Eng Des Sel 23 289-97 (2010); Gong R et al., PLoS ONE 7: e42288 (2012); Wozniak-Knopp G et al., PLoS ONE 7: e30083 (2012); Ying T et al., J Biol Chem 287: 19399-408 (2012); Ying T et al., J Biol Chem 288: 25154-64 (2013); Chiang M et al., J Am Chem Soc 136: 3370-3 (2014); Rader C, Trends Biotechnol 32: 186-97 (2014); Ying T et al., Biochimica Biophys Acta 1844: 1977-82 (2014).
[0267] In accordance with certain other embodiments, the binding region includes engineered, alternative scaffolds to immunoglobulin domains that exhibit similar functional characteristics, such as high-affinity and specific binding of target biomolecules, and enables the engineering of improved characteristics, such as greater stability or reduced immunogenicity. For certain embodiments of the cell-targeting molecules of the present the invention, the binding region is selected from the group which includes engineered Armadillo repeat polypeptides (ArmRPs), engineered, fibronectin-derived, 10.sup.th fibronectin type III (10Fn3) domain (monobodies, AdNectins, or AdNexins); engineered, tenascin-derived, tenascin type III domain (Centryns); engineered, ankyrin repeat motif containing polypeptide (DARPins); engineered, low-density-lipoprotein-receptor-derived, A domain (LDLR-A) (Avimers); lipocalin (anticalins); engineered, protease inhibitor-derived, Kunitz domain; engineered, Protein-A-derived, Z domain (Affibodies); engineered, gamma-B crystallin-derived scaffold or engineered, ubiquitin-derived scaffold (Affilins); Sac7d-derived polypeptides (Nanoffitins or affitins); engineered, Fyn-derived, SH2 domain (Fynomers); and engineered antibody mimic and any genetically manipulated counterparts of the foregoing that retains its binding functionality (Worn A, Pluckthun A, J Mol Biol 305: 989-1010 (2001); Xu L et al., Chem Biol 9: 933-42 (2002); Wikman M et al., Protein Eng Des Sel 17: 455-62 (2004); Binz H et al., Nat Biotechnol 23: 1257-68 (2005); Holliger P, Hudson P, Nat Biotechnol 23: 1126-36 (2005); Gill D, Damle N, Curr Opin Biotech 17: 653-8 (2006); Koide A, Koide S, Methods Mol Biol 352: 95-109 (2007); Byla P et al., J Biol Chem 285: 12096 (2010); Zoller F et al., Molecules 16: 2467-85 (2011); Alfarano P et al., Protein Sci 21: 1298-314 (2012); Madhurantakam C et al., Protein Sci 21: 1015-28 (2012); Varadamsetty G et al., J Mol Biol 424: 68-87 (2012)).
[0268] Among certain embodiments of the present invention, the immunoglobulin-type binding region is derived from a nanobody or single domain immunoglobulin-derived region V.sub.HH. Generally, nanobodies are constructed from fragments of naturally occurring single, monomeric variable domain antibodies (sdAbs) of the sort found in camelids and cartilaginous fishes (Chondrichthyes). Nanobodies are engineered from these naturally occurring antibodies by truncating the single, monomeric variable domain to create smaller and more stable molecules, such as, e.g., IgNAR, V.sub.HH, and V.sub.NAR constructs. Due to their small size, nanobodies are able to bind to antigens that are not accessible to whole antibodies.
[0269] In certain embodiments of the cell-targeting molecule of the present invention, the binding region comprises a polypeptide(s) selected from the group consisting of: a) a heavy chain variable (V.sub.H) domain comprising (i) a HABR1 or HCDR1, (ii) a HABR2 or HCDR2, and (iii) a HABR3 or HCDR3; and/or b) a light chain variable (V.sub.L) domain comprising (i) a LABR1 or LCDR1, (ii) a LABR2 or LCDR2, and (iii) a LABR3 or LCDR3; wherein each of the aforementioned ABRs and/or CDRs is selected from the polypeptide comprising or consisting essentially of one of the amino acid sequence as shown in SEQ ID NOs: 844-1100. In certain further embodiments, the binding region comprises or consists essentially of 269-499 of any one of SEQ ID NOs: 807-808 and 812-813, comprises or consists essentially of amino acids of 269-519 of any one of SEQ ID NOs: 814-815 and 818-829, or comprises or consists essentially of amino acids 268-386 of any one of SEQ ID NOs: 816-817.
[0270] In certain embodiments of the present invention, the binding region polypeptide comprises a free cysteine residue suitable for conjugation to another molecule, wherein the cysteine is at or proximal to a carboxy-terminus of the binding region polypeptide. In certain further embodiments, the binding region of a cell-targeting molecule of the present invention comprises a series of amino acid residues represented by (X)n-C-X or (X)n-C, where X refers to any amino acid, (X)n refers to a polypeptide comprising a binding domain, and C refers to a cysteine residue.
[0271] In certain embodiments of the present invention, the binding region polypeptide comprises a free cysteine residue suitable for conjugation to another molecule, wherein the cysteine is at or proximal to an amino-terminus of the binding region polypeptide. In certain further embodiments, the binding region of a cell-targeting molecule of the present invention comprises a series of amino acid residues represented by C-(X)n or M-C-(X)n, where X refers to any amino acid, (X)n refers to a polypeptide comprising a binding domain, C refers to a cysteine residue, and M refers to a starting methionine.
[0272] In certain embodiments of the present invention, the molecule of the present invention comprises an immunoglobulin binding region which lacks cysteine residues. Such immunoglobulin binding region structures are known to the skilled worker and/or can be created using routine methods (see e.g. Proba K, J Mol Biol 275: 245-53 (1998)).
[0273] Any of the above binding regions may be used as a component of the cell-targeting molecules of the present invention as long as the binding region component has a dissociation constant of 10.sup.5 to 10.sup.12 moles per liter, preferably less than 200 nanomolar (nM), towards an extracellular target biomolecule.
[0274] Cell-specific targeting can be accomplished by attaching molecules of the present invention to cell targeting carriers, such as, e.g., liposomes, polymers, nanocarriers, microspheres, nanospheres, dendrimers, polymeric micelles, silicon or carbon materials, such as e.g., nanotubes, nanorods and nanohorns, magnetic nanoparticles, microemulsions, and other nanostructures (Sinha R et al., Molecular Cancer Therapeutics 5: 1909-17 (2006); L Brinton et al., Journal of the National Cancer Institute 100: 1643-8 (2008); Tanaka T et al., Biomed Micro Devices 11: 49-63 (2009)). Attachment may be accomplished using one or more covalent bonds and/or encapsulation.
Extracellular Target Biomolecules
[0275] The binding region of the molecule of the invention comprises a polypeptide region capable of binding specifically to an extracellular target biomolecule, preferably which is physically-coupled to the surface of a cell type of interest, such as a cancer cell, tumor cell, plasma cell, infected cell, or host cell harboring an intracellular pathogen.
[0276] The term target biomolecule refers to a biological molecule, commonly a protein or a protein modified by post-translational modifications, such as glycosylation, which is capable of being bound by a binding region to target a protein to a specific cell-type or location within an organism. Extracellular target biomolecules may include various epitopes, including unmodified polypeptides, polypeptides modified by the addition of biochemical functional groups, and glycolipids.
[0277] There are numerous extracellular target biomolecules known to the skilled worker that may be targeted by the binding region of a cell-targeting molecule of the present invention and polypeptide binding domains known to bind such target biomolecules (see e.g. WO 2014/164680; WO 2014/164693; WO 2015/113005; WO 2015/113007; WO 2015/138435; WO 2015/138452; US20150259428; WO 2015/191764; US20160177284; WO 2016/126950).
[0278] For purposes of the present invention, the term extracellular with regard to modifying a target biomolecule refers to a biomolecule that has at least a portion of its structure exposed to the extracellular environment. Extracellular target biomolecules include cell membrane components, transmembrane spanning proteins, cell membrane-anchored biomolecules, cell-surface-bound biomolecules, and secreted biomolecules.
[0279] With regard to the present invention, the phrase physically coupled when used to describe a target biomolecule means both covalent and/or non-covalent intermolecular interactions that couple the target biomolecule, or a portion thereof, to the outside of a cell, such as a plurality of non-covalent interactions between the target biomolecule and the cell where the energy of each single interaction is on the order of about 1-5 kiloCalories (e.g. electrostatic bonds, hydrogen bonds, Van der Walls interactions, hydrophobic forces, etc.). All integral membrane proteins can be found physically coupled to a cell membrane, as well as peripheral membrane proteins. For example, an extracellular target biomolecule might comprise a transmembrane spanning region, a lipid anchor, a glycolipid anchor, and/or be non-covalently associated (e.g. via non-specific hydrophobic interactions and/or lipid binding interactions) with a factor comprising any one of the foregoing.
[0280] The binding regions of the cell-targeting molecules of the present invention may be designed or selected based on numerous criteria, such as the cell-type specific expression of their target biomolecules and/or the physical localization of their target biomolecules with regard to specific cell types. For example, certain cell-targeting molecules of the present invention comprise binding domains capable of binding cell-surface targets which are expressed exclusively by only one cell-type to the cell surface.
[0281] Among certain embodiments of the present invention, the cell-targeting molecule comprises a binding region derived from an immunoglobulin-type polypeptide selected for specific and high-affinity binding to a surface antigen on the cell surface of a cancer cell, where the antigen is restricted in expression to cancer cells (see Glokler J et al., Molecules 15: 2478-90 (2010); Liu Y et al., Lab Chip 9: 1033-6 (2009)). In accordance with other embodiments, the binding region is selected for specific and high-affinity binding to a surface antigen on the cell surface of a cancer cell, where the antigen is over-expressed or preferentially expressed by cancer cells as compared to non-cancer cells. Some representative target biomolecules include, but are not limited to, the following enumerated targets associated with cancers and/or specific immune cell types.
[0282] Many immunoglobulin-type binding regions that recognize epitopes associated with cancer cells exist in the prior art, such as binding regions that target annexin AI, B3 melanoma antigen, B4 melanoma antigen, B7-H3 (CD276, B7RP-2), B-cell maturation antigen (BCMA, BCM, TNRSF17, CD269), CD2, CD3, CD4, CD19, CD20 (B-lymphocyte antigen protein CD20), CD22, CD25 (interleukin-2 receptor IL2R), CD30 (TNFRSF8), CD37, CD38 (cyclic ADP ribose hydrolase), CD40, CD44 (hyaluronan receptor), protein tyrosine phosphatase receptor type C (CD45, PTPRC, LCA), ITGAV (CD51), CD56, CD66, CD70, CD71 (transferrin receptor), CD73, CD74 (HLA-DR antigens-associated invariant chain), CD79 (e.g. CD79a or CD79b), CD98, endoglin (END, CD105), CD106 (VCAM-1), CD138, chemokine receptor type 4 (CDCR-4, fusin, CD184), CD200, insulin-like growth factor 1 receptor (CD221), mucin1 (MUC1, CD227, CA6, CanAg), basal cell adhesion molecule (B-CAM, CD239), CD248 (endosialin, TEM1), tumor necrosis factor receptor 10b (TNFRSF10B, CD262), tumor necrosis factor receptor 13B (TNFRSF13B, TACI), vascular endothelial growth factor receptor 2 (KDR, CD309), epithelial cell adhesion molecule (EpCAM, CD326), human epidermal growth factor receptor 2 (HER2, Neu, ErbB2, CD340), cancer antigen 15-3 (CA15-3), cancer antigen 19-9 (CA 19-9), cancer antigen 125 (CA125, MUC16), CA242, carcinoembryonic antigen-related cell adhesion molecules (e.g. CEACAM3 (CD66d) and CEACAM5), carcinoembryonic antigen protein (CEA), choline transporter-like protein 4 (SLC44A4), chondroitin sulfate proteoglycan 4 (CSP4, MCSP, NG2), CTLA4, delta-like proteins (e.g. DLL3, DLL4), ectonucleotide pyrophosphatase/phosphodiesterase proteins (e.g. ENPP3), endothelin receptors (ETBRs), epidermal growth factor receptor (EGFR, ErbB1), Epstein-Barr virus latent membrane protein 1 (LMP1), folate receptors (FOLRs, e.g. FR), G-28, ganglioside GD2, ganglioside GD3, HLA-Dr10, HLA-DRB, human epidermal growth factor receptor 1 (HER1), HER3/ErbB-3, Ephrin type-B receptor 2 (EphB2), epithelial cell adhesion molecule (EpCAM), fibroblast activation protein (FAP/seprase), guanylyl cyclase c (GCC), insulin-like growth factor 1 receptor (IGF1R), interleukin 2 receptor (IL-2R), interleukin 6 receptor (IL-6R), integrins alpha-V beta-3 (.sub.V.sub.3), integrins alpha-V beta-5 (v5), integrins alpha-5 beta-1 (.sub.5.sub.1), L6, zinc transporter (LIV-1), MPG, melanoma-associated antigen 1 protein (MAGE-1), melanoma-associated antigen 3 (MAGE-3), mesothelin (MSLN), metalloreductase STEAP1, MPG, MS4A, NaPi2b, nectins (e.g. nectin-4), p21, p97, polio virus receptor-like 4 (PVRL4), protease-activated-receptors (such as PAR1), prostate-specific membrane antigen proteins (PSMAs), SAIL (C16orf54), SLIT and NTRK-like proteins (e.g. SLITRK6), Thomas-Friedenreich antigen, transmembrane glycoprotein (GPNMB), trophoblast glycoproteins (TPGB, 5T4, WAIF1), and tumor-associated calcium signal transducers (TACSTDs, e.g. Trop-2, EGP-1, etc) (see e.g. Lui B et al., Cancer Res 64: 704-10 (2004); Novellino L et al., Cancer Immunol Immunother 54: 187-207 (2005); Bagley R et al., Int J Oncol 34: 619-27 (2009); Gerber H et al., mAbs 1: 247-53 (2009); Beck A et al., Nat Rev Immunol 10: 345-52 (2010); Andersen J et al., J Biol Chem 287: 22927-37 (2012); Nolan-Stevaux O et al., PLoS One 7: e50920 (2012); Rust S et al., Mol Cancer 12: 11 (2013); Kim S et al., Blood Cancer Journal 5: e316 (2015)). This list of target biomolecules is intended to be non-limiting. It will be appreciated by the skilled worker that any desired target biomolecule associated with a cancer cell or other desired cell type may be used to design or select a binding region to be coupled with a toxin effector polypeptide to produce a cell-targeting molecule of the present invention.
[0283] Examples of other target biomolecules which are strongly associated with cancer cells and immunoglobulin-type binding regions known to bind them include BAGE proteins (B melanoma antigens), basal cell adhesion molecules (BCAMs or Lutheran blood group glycoproteins), bladder tumor antigen (BTA), cancer-testis antigen NY-ESO-1, cancer-testis antigen LAGE proteins, CD19 (B-lymphocyte antigen protein CD19), CD21 (complement receptor-2 or complement 3d receptor), CD26 (dipeptidyl peptidase-4, DPP4, or adenosine deaminase complexing protein 2), CD33 (sialic acid-binding immunoglobulin-type lectin-3), CD52 (CAMPATH-1 antigen), CD56, CS1 (SLAM family number 7 or SLAMF7), cell surface A33 antigen protein (gpA33), Epstein-Barr virus antigen proteins, GAGE/PAGE proteins (melanoma associated cancer/testis antigens), hepatocyte growth factor receptor (HGFR or c-Met), MAGE proteins, melanoma antigen recognized by T-cells 1 protein (MART-1/MelanA, MARTI), mucins, Preferentially Expressed Antigen of Melanoma (PRAME) proteins, prostate specific antigen protein (PSA), prostate stem cell antigen protein (PSCA), Receptor for Advanced Glycation Endroducts (RAGE), tumor-associated glycoprotein 72 (TAG-72), vascular endothelial growth factor receptors (VEGFRs), and Wilms' tumor antigen.
[0284] Examples of other target biomolecules which are strongly associated with cancer cells are carbonic anhydrase IX (CA9/CAIX), claudin proteins (CLDN3, CLDN4), ephrin type-A receptor 3 (EphA3), folate binding proteins (FBP), ganglioside GM2, insulin-like growth factor receptors, integrins (such as CD11a-c), receptor activator of nuclear factor kappa B (RANK), receptor tyrosine-protein kinase erB-3, SAIL (C16orf54), tumor necrosis factor receptor 10A (TRAIL-R1/DR4), tumor necrosis factor receptor 10B (TRAIL-R2), tenascin C, and CD64 (FcRI) (see Hough C et al., Cancer Res 60: 6281-7 (2000); Thepen T et al., Nat Biotechnol 18: 48-51 (2000); Pastan I et al., Nat Rev Cancer 6: 559-65 (2006); Pastan, Annu Rev Med 58: 221-37 (2007); Fitzgerald D et al., Cancer Res 71: 6300-9 (2011); Scott A et al., Cancer Immun 12: 14-22 (2012); Kim S et. al., Blood Cancer Journal 5: e316 (2015)). This list of target biomolecules is intended to be non-limiting.
[0285] In addition, there are numerous other examples of contemplated, target biomolecules such as ADAM metalloproteinases (e.g. ADAM-9, ADAM-10, ADAM-12, ADAM-15, ADAM-17), ADP-ribosyltransferases (ART1, ART4), antigen F4/80, bone marrow stroma antigens (BST1, BST2), break point cluster region-c-abl oncogene (BCR-ABL) proteins, C3aR (complement component 3a receptors), CD7, CD13, CD14, CD15 (Lewis X or stage-specific embryonic antigen 1), CD23 (FC epsilon RH), CD49d, CD53, CD54 (intercellular adhesion molecule 1), CD63 (tetraspanin), CD69, CD80, CD86, CD88 (complement component 5a receptor 1), CD115 (colony stimulating factor 1 receptor), IL-1R (interleukin-1 receptor), CD123 (interleukin-3 receptor), CD129 (interleukin 9 receptor), CD183 (chemokine receptor CXCR3), CD191 (CCR1), CD193 (CCR3), CD195 (chemokine receptor CCR5), CD203c, CD225 (interferon-induced transmembrane protein 1), CD244 (Natural Killer Cell Receptor 2B4), CD282 (Toll-like receptor 2), CD284 (Toll-like receptor 4), CD294 (GPR44), CD305 (leukocyte-associated immunoglobulin-like receptor 1), ephrin type-A receptor 2 (EphA2), FceRIa, galectin-9, alpha-fetoprotein antigen 17-A1 protein, human aspartyl (asparaginyl) beta-hydroxylase (HAAH), immunoglobulin-like transcript ILT-3, lysophosphatidlglycerol acyltransferase 1 (LPGAT1/IAA0205), lysosome-associated membrane proteins (LAMPs, such as CD107), melanocyte protein PMEL (gp100), myeloid-related protein-14 (mrp-14), NKG2D ligands (e.g., MICA, MICB, ULBP1, ULBP2, UL-16-binding proteins, H-60s, Rae-1s, and homologs thereof), receptor tyrosine-protein kinase erbB-3, SART proteins, scavenger receptors (such as CD64 and CD68), Siglecs (sialic acid-binding immunoglobulin-type lectins), syndecans (such as SDC1 or CD138), tyrosinase, tyrosinease-related protein 1 (TRP-1), tyrosinease-related protein 2 (TRP-2), tyrosinase associated antigen (TAA), APO-3, BCMA, CD2, CD3, CD4, CD8, CD18, CD27, CD28, CD29, CD41, CD49, CD90, CD95 (Fas), CD103, CD104, CD134 (OX40), CD137 (4-1BB), CD152 (CTLA-4), chemokine receptors, complement proteins, cytokine receptors, histocompatibility proteins, ICOS, interferon-alpha, interferon-beta, c-myc, osteoprotegerin, PD-1, RANK, TACI, TNF receptor superfamily member (TNF-R1, TNFR-2), Apo2/TRAIL-R1, TRAIL-R2, TRAIL-R3, and TRAIL-R4 (see Scott A et al., Cancer Immunity 12: 14 (2012); Cheever M et al., Clin Cancer Res 15: 5323-37 (2009)), for target biomolecules and note the target molecules described therein are non-limiting examples). It will be appreciated by the skilled worker that any desired target biomolecule may be used to design or select a binding region to be coupled with a toxin effector polypeptide to produce a cell-targeting molecule of the present invention.
[0286] In certain embodiments, the binding region comprises or consists essentially of an immunoglobulin-type polypeptide selected for specific and high-affinity binding to the cellular surface of a cell type of the immune system. For example, immunoglobulin-type binding domains are known that bind to programmed death ligand 1 (PD-L1), CD1, CD2, CD3, CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11, CD12, CD13, CD14, CD15, CD16, CD17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD33, CD34, CD35, CD36, CD37, CD38, CD40, CD41, CD56, CD61, CD62, CD66, CD95, CD117, CD123, CD235, CD146, CD326, interleukin-2 receptor (IL-2R), receptor activator of nuclear factor kappa B (RANKL), SLAM-associated protein (SAP), and TNFSF18 (tumor necrosis factor ligand 18 or GITRL).
[0287] Extracellular target biomolecules of the binding region of the cell-targeting molecules of the present invention may include biomarkers over-proportionately or exclusively present on cancer cells, immune cells, and cells infected with intracellular pathogens, such as viruses, bacteria, fungi, prions, or protozoans.
[0288] This general structure is modular in that any number of diverse cell-targeting binding regions may be linked to one or more toxin effector polypeptides to produce a cell-targeting molecule of the present invention, and compositions thereof.
C. Toxin Effector Polypeptides Derived from Toxins
[0289] In certain embodiments, the cell-targeting molecule of the present invention comprises a toxin effector polypeptide region derived from a proteinaceous toxin other than a Shiga toxin(s). In certain embodiments, the cell-targeting molecule of the present invention does not comprise a Shiga toxin effector polypeptide. In certain embodiments, the cell-targeting molecule of the present invention comprises a toxin effector region derived from a toxin other than a member of the Shiga toxin family, such as, e.g., from an ABx toxin other than a Shiga toxin, a ribosome inactivating protein toxin other than Shiga toxin, abrin, anthrax toxin, Aspfl, bouganin, bryodin, cholix toxin, claudin, diphtheria toxin, gelonin, heat-labile enterotoxin, mitogillin, pertussis toxin, pokeweed antiviral protein, pulchellin, Pseudomonas exotoxin A, restrictocin, ricin, saporin, sarcin, and subtilase cytotoxin (see e.g., WO 2015/113005; WO 2015/120058). In certain embodiments, the cell-targeting molecule of the present invention does not comprise either a toxin effector region or any polypeptide derived from a toxin.
[0290] In certain embodiments, the cell-targeting molecule of the present invention comprises a toxin effector polypeptide which is not a Shiga toxin effector polypeptide. The present invention contemplates the use of various polypeptides derived from toxins as toxin effector polypeptides. For example, many toxins represent optimal sources of cytotoxic polypeptides and/or proteasome delivering effector polypeptides because of the wealth of knowledge about their intracellular routing behaviors (see e.g. WO 2015/113005). Any protein toxin with the intrinsic ability to intracellularly route from an early endosomal compartment to either the cytosol or the ER represents a source for a proteasome delivery effector polypeptide which may be exploited for the purposes of the present invention, such as a starting component for modification or as a source for mapping a smaller proteasome delivery effector region therein.
[0291] In certain embodiments, the cell-targeting molecule of the present invention comprises a toxin effector lacking any lysine residues or comprising exactly one lysine residue for site-specific conjugation. The catalytic domains of many toxins, especially toxins which use retrograde routing pathways via the endoplasmic reticulum, are devoid of lysine residues (DeLange R et. al., Proc Natl Acad Sci USA 73: 69-72 (1976); London E, Luongo C, Biochem Biophys Res Commun 160: 333-9 (1989); Hazes B, Read R, Biochemistry 36: 11051-4 (1997); Deeks E et al., Biochemistry 41: 3405-13 (2002); Worthington Z, Carbonetti N, Infect Immun 75: 2946-53 (2007)).
[0292] In certain embodiments, the cell-targeting molecule of the present invention comprises the toxin effector polypeptide which is a Shiga toxin effector polypeptide. In certain further embodiments, the Shiga toxin effector polypeptide is a SLT-1A-Cys1-variant (e.g. SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, and 75). In certain further embodiments, the Shiga toxin effector polypeptide is carboxy-terminal to another proteinaceous component of the cell-targeting molecule (e.g. a cell-targeting binding region or starting methionine) such that the cysteine residue at position 1 of the Shiga toxin effector polypeptide (C1) forms a peptide bond via its amino group.
[0293] The general structure of the cell-targeting molecules of the present invention is modular, in that various, diverse binding regions may be used with the same toxin effector polypeptide (e.g. a Shiga toxin effector polypeptide of the present invention) to provide for diverse targeting of various extracellular target biomolecules and thus targeting of cytotoxicity, cytostasis, and/or exogenous material delivery to various diverse cell types. In the cell-targeting molecules of the invention, the binding region(s) and Shiga toxin effector polypeptide(s) may be directly linked to each other and/or suitably linked to each other via one or more linkers well known in the art. For the purposes of the cell-targeting molecules of the present invention, the specific order or orientation is not fixed for the toxin effector polypeptide and the cell-targeting, binding region in relation to each other (see e.g.
D. Amino Acid Residues and Structures Amenable for Site-Specific Conjugation
[0294] Any amino acid residue having a bio-orthogonal reactive moiety (e.g. a side chain or functional group) may be suitable as conjugation site in the molecules of the present invention. The skilled worker can select an amenable and suitable amino acid from the prior art or using routine techniques can identify through experimentation a novel amino acid which is amenable and suitable for use in a molecule of the present invention. In certain embodiments, a molecule of the present invention comprises a Shiga toxin effector polypeptide having a unique amino acid residue, which may be either ectopically positioned or naturally occurring at that position. In certain other embodiments, the invention may involve a unique amino acid or a non-unique amino acid residue which is uniquely accessible, i.e. not buried in the interior of a proteinaceous structure as are all other) residues of the same amino acid type.
[0295] In certain embodiments of the present invention, a single lysine, histidine, or cysteine is engineered into a cell-targeting molecule to provide a specific residue site for conjugation of another molecule. In general, the strongly nucleophilic functional groups of the amino acids lysine, histidine, and cysteine make these three amino acid residues the most amenable attachment points for chemically conjugating of an atom or molecule to a protein. However, the number of lysines, histidines, and/or cysteines vary from protein to protein, which affects the suitability of site-specific attachment and/or control of conjugate stoichiometry. In the examples below, all the cysteine residues were removed from a parental, Shiga toxin A Subunit polypeptide. Then, an ectopic cysteine residue was engineered into the polypeptide to provide a unique attachment position for controlled conjugation. This was repeated at different positions to provide a group of cysteine-engineered Shiga toxin effector polypeptides, and these polypeptides are tested for retention of Shiga toxin effector functions and ability to deliver a conjugated molecule to the inside of a cell.
[0296] In certain embodiments of the present invention, a single, unnatural amino acid residue is engineered into a cell-targeting molecule to provide a specific residue site for conjugation of another molecule (see e.g. Liu W et al., Nat Methods 4: 239-44 (2007); Liu C, Schultz P, Annu Rev Biochem 79: 413-44 (2010); Young T et al., J Mol Biol 395: 361-74 (2010); Young T, Schultz P, J Biol Chem 285: 11039-44 (2010); Young D et al., Biochemistry 50: 1894-900 (2011); Hoesl M, Budisa N, Curr Opin Biotechnol 23: 751-7 (2012); Ozawa K et al., Biochem Biophys Res Commun 418: 652-6 (2012); Chin J, Annu Rev Biochem 83: 379-408 (2014); Ozawa K, Loh C, Methods Mol Biol 1118: 189-203 (2014)). For example, unnatural amino acid residues such as selenocysteine and para-acetylphenylalanine, are known in the art to provide useful conjugation sites. Similarly, amino acid residues having an azide group can be used for site specific conjugation.
[0297] In certain embodiments of the present invention, the site-specific attachment site is the primary amine or carboxy-terminal of a polypeptide component of the cell-targeting molecule of the present invention.
[0298] In certain embodiments of the present invention, a short polypeptide motif is engineered into a cell-targeting molecule to provide a specific residue site for conjugation of another molecule. For example, cysteine residues in certain motifs like CxPxR can be modified by formylglycine generating enzymes into formylglycine, and then the resulting aldehyde functional group may be conjugated to another molecule using hydrozino-Pictet-Spengler chemistry (see e.g. Carrico I et al., Nat Chem Biol 3: 321-2 (2007); Rabuka D, et al., Nat Protoc 7: 1052-67 (2012)).
[0299] It is important to note that the solvent accessibility of the engineered residue may affect its ability to provide a suitable conjugation site; however, suitability may vary with the conjugated molecule and specific application of the resulting conjugate. For example, the size and steric hindrances of the conjugated molecule may affect the stability of the final conjugated product such that certain engineered sites and/or ectopic residues are suitable for certain conjugated molecules but not others. While highly solvent accessible amino acid residues may be predicted to provide better sites for conjugation, it is important to note that the Shiga toxin effector polypeptide or cell-targeting molecule with the greatest therapeutic utility may require a conjugation site which is less solvent accessible than other possible conjugation sites (see e.g. Shen B et al., Nat Biotechnol 30: 184-9 (2012)).
[0300] There are several common conjugation strategies for linking a proteinaceous molecule to another molecule, such as, e.g., via a lysine in a protein to an amine-reactive linker or molecule, via a cysteine in a protein to a sulfhydryl reactive linker or molecule (e.g. involving an activated maleimide group), via enzyme-mediated conjugation, and/or via the incorporation of an unnatural amino acid such as para-acetylphenylalanine (see e.g. Kline T et al., Pharm Res 32: 3480-93 (2015)). For example, sulfhydryl-reactive chemical groups are highly amenable to conjugation chemistry, such as via alkylation (usually the formation of a thioether bond) or disulfide exchange (formation of a disulfide bond). Non-limiting examples of sulfhydryl-reactive groups include haloacetyls, maleimides, aziridines, acryloyls, arylating agents, vinylsulfones, pyridyl disulfides, TNB-thiols and disulfide reducing agents. For example, a cysteine residue can be conjugated to another molecule, e.g., using maleimide or bromoacetamide groups on linkers or cargos and/or using click chemistry. Unnatural amino acid residues can be conjugated using oxime groups on linkers or cargos and/or via click chemistry. For example, para-acetylphenylalanine can be conjugated to another molecule comprising an alkoxy-amine via oxime ligation. For example, selenocysteine can be conjugated to another molecule using maleimide groups and/or click chemistry (see e.g. Hofer T et al., Biochemistry 48: 12047-57 (2009); Young T et al., J Mol Biol 395: 361-74 (2010); Kiick K et al., Proc Natl Acad Sci USA 99: 19-24 (2002)).
[0301] For certain embodiments, the molecule of the present invention is made using a haloalkyl derivative, such as, e.g., an iodoacetamide or maleimide, present in the cargo or linker to link the cargo to a cysteine, methionine, and/or histidine residue(s) present in a polypeptide component of a molecule of the present invention. In certain embodiments, a maleimide agent is specifically used to avoid conjugation to a tyrosine, histidine, and/or methionine residue(s).
[0302] A cysteine residue already present in a polypeptide component of a molecule of the present invention may be used, such as, e.g., after a reduction reaction to make free its thiol group and make it available for conjugation. The skilled worker can reduce a thiol linkage using methods known in the art, such as, e.g., using TCEP (tris(2-carboxyethyl)phosphine hydrochloride, dithiothreitol (DTT), and/or beta-mercaptoethanol (BME).
[0303] For certain embodiments, the molecule of the present invention is made using a nitrosylated thiol derivative, such as, e.g., a thiosulfate, present in the cargo or linker to link the cargo to a cysteine, methionine, and/or histidine residue(s) present in a polypeptide component of a molecule of the present invention.
[0304] For certain embodiments, a homobifunctional maleimide, homobifunctional sulfhydryl-reactive maleimide, heterobifunctional maleimide, and/or heterobifunctional amine-to-sulfhydryl maleimide crosslinker(s).
[0305] For certain embodiments, a haloacetyl reagent is used, such as, e.g., a haloacetyl crosslinker contain an iodoacetyl or a bromoacetyl group and/or using a NHS ester amine- to sulfhydryl crosslinker. For certain embodiments, an iodoacetyl reactive group is used for chemical conjugation to a sulfhydryl group, such as, e.g., a homobifunctional or heterobifunctional iodoacetyl crosslinker. For certain embodiments, a bromoacetyl reactive group is used for chemical conjugation to a sulfhydryl group, such as, e.g., a homobifunctional or heterobifunctional bromoacetyl crosslinker.
[0306] For certain embodiments, a sulfhydryl group is added using a chemical reaction, such as, e.g., using 2-iminothiolane, SATA, SATP, SAT(PEG).sub.4, or a pyridyl disulfide. In certain embodiments, the cell-targeting molecule is treated with Traut's Reagent (2-iminothiolane, 2-IT) or SATA to add sulfhydryl groups onto primary amine sites.
[0307] In certain embodiments, the linking to the Shiga toxin effector polypeptide involves a chemical reaction involving a sulfhydryl group, such as, e.g., a sulfhydryl group of a cysteine, methionine, N-formylmethionine, homocysteine, or taurine residue of the Shiga toxin effector polypeptide, the linker connecting the Shiga toxin effector polypeptide to a cell-targeting binding region, or the cargo being conjugated.
[0308] Chemical reactions suitable for use in conjugating an atom or molecule to a polypeptide or polypeptide component of the present invention include: carbodiimide-mediated reaction, EDC-mediated amide bond formation, hydrazide activation reaction, pyridyl disulfide reaction, iodoacetyl reaction, and/or the Mannich reaction.
[0309] For example, a carboxyl group of a polypeptide or polypeptide component of the present invention may be linked to a cargo via a carbodiimide-mediated reaction, such as, e.g., using a water-soluble carbodiimide crosslinker, that results in amide bond formation with an amino, amine, and/or hydrazine group of the cargo. The carboxyl group of the polypeptide may be part of a surface accessible amino acid residue, such as, e.g., the carboxy-terminal residue, an aspartic acid, and/or glutamic acid.
[0310] For examples, EDC-mediated amide bond formation may be used for the linking of a carboxylate group of a cargo with the primary amine group of the amino-terminal amine group of a polypeptide or polypeptide component of the present invention. Alternatively, EDC-mediated amide bond formation may be used for the linking of an amine group of a cargo with the carboxy-terminal carboxylate group of a polypeptide or polypeptide component of the present invention. The skilled worker may then use a purification step(s) to further isolate and purify the desired conjugate molecule(s).
[0311] For example, the Mannich reaction may be used for the condensation of an aldehyde group of a cargo with the active hydrogen of an amine group of an amino acid residue, such as, e.g., an amine group of an arginine, histidine, lysine, asparagine, glutamine, proline, or tryptophan residue, and/or the amino-terminal amine group of a polypeptide or polypeptide component of the present invention. The skilled worker may then use a purification step(s) to further isolate and purify the desired conjugate molecule(s).
[0312] For example, a pyridyl disulfide of a cargo or linker may be used to react with a sulfhydryl group of a polypeptide or polypeptide component of the present invention. The pyridyl disulfide may be pre-activated via other chemical reactions before the coupling reaction between the cargo and the protein or polypeptide component of the present invention.
[0313] For example, an iodoacetyl of a cargo may be used to react with a sulfhydryl group of a polypeptide or polypeptide component of the present invention. The pyridyl disulfide may be pre-activated via other chemical reactions before the coupling reaction between the cargo and the protein or polypeptide component of the present invention.
[0314] In certain embodiments, the cargo and Shiga toxin effector polypeptide of the present invention are linked via a disuccinimidyl suberate (DSS) linker.
E. Linkers for Conjugating Polypeptide Components of the Present Invention to a Heterologous Molecule, Cargo, and/or Additional Exogenous Material
[0315] In certain embodiments, the molecule of the present invention comprises a conjugated molecule, such as, e.g., a heterologous molecule, a cargo, an additional exogenous material, and/or a cell-targeting molecule altering agent. In certain further embodiments, the molecule of the present invention is indirectly conjugated via one or more linkers, referred to herein as a conjugation linker. The skilled worker can select conjugation linker from the prior art or using routine techniques can identify through experimentation a novel conjugation linker suitable for use in a molecule of the present invention. In certain embodiments, the conjugation linkage involves an amine reactive compound and/or a sulfhydryl reactive compound, such as, e.g., a thiol.
[0316] Conjugation linkers are generally those which allow stable linkage of the conjugate molecule to a Shiga toxin effector polypeptide and/or proteinaceous component of the cell-targeting molecule of the present invention. Stability in this sense refers to both during storage and in the circulatory system of a vertebrate; however, in certain embodiments the conjugation linker allows for the selective dissociation of the conjugate molecule from the linker and/or the remainder of the cell-targeting molecule, and/or the degradation of the linker thereby breaking the linkage between the conjugate molecule and the remainder of the cell-targeting molecule.
[0317] For example, suitable conjugation linkers known to the skilled worker include or involve hydrazone linkages, thio-ether linkages, disulfide linkages, proteolytically-cleaved linkers, valine-citrulline linkers, -glucuronide linkers, self-immolative linkages, reversible amino-thiol linkages, and SpaceLink linkers (see e.g. Hjfeldt J et al., J Org Chem 71: 9556-9 (2006); Ducry L, Stump B, Bioconjugate Chem 21: 5-13 (2010)).
[0318] In certain embodiments, a molecule of the present invention comprises the conjugation linker which is homobifunctional, whereas in other embodiments, the conjugation linker is heterobifunctional. Examples of homobifunctional linkers suitable for use in certain embodiments of the present invention include NHS esters, haloacetyl, aryl azide, diazirine, imidoesters, carbodiimide, maleimide, hydrazide, pyridyldithiol, isocyanate, psoralen. In certain embodiments, the conjugation linker comprises a polyethylene glycol (PEG) spacer. Examples of homobifunctional and/or heterobifunctional linkers suitable for use in certain embodiments of the present invention include bis(sulfosuccinimidyl) suberate, disuccinimidyl suberate, bis(succinimidyl) penta (ethylene glycol), bis(succinimidyl) nona (ethylene glycol), m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester, 2-pyridyldithiol-tetraoxaoctatriacontane-N-hydrosuccinimide, and succinimidyl-[(N-maleimidopropionamido)-tetracosaethyleneglycol]ester.
[0319] In certain embodiments, a molecule of the present invention comprises a maleimide type conjugation linker. In certain further embodiments, a molecule of the present invention comprises a dibenzocyclooctyne (DBCO) maleimide linker linking a cargo molecule to a proteinaceous component of the cell-targeting molecule, such as, e.g., via a cysteine residue in the proteinaceous component.
[0320] In certain embodiments, the conjugation linker is conjugated to another molecule via a disulfide bond(s) having a sterically-hindered carbon(s) near the sulfur atom(s) (see e.g. Erickson H et al., Cancer Res 66: 4426-33 (2006)).
[0321] In certain embodiments, the conjugation linker is a disulfide type linker (see e.g. Hamilton G, Biologicals 43: 318-32 (2015)).
[0322] In certain embodiments of the present invention, the components of a Shiga toxin effector polypeptide (e.g. a cargo or to a Shiga toxin effector polypeptide) or cell-targeting molecule (e.g. a binding region to a toxin effector polypeptide) may be suitably linked to each other via one or more linkers well known in the art and/or described herein, such as, e.g., proteinaceous linkers capable of being genetically fused between other proteinaceous components of the cell-targeting molecules of the present invention.
[0323] Suitable linkers are generally those which allow each polypeptide component of the present invention to fold with a three-dimensional structure very similar to the polypeptide components produced individually without any linker or other component. Suitable linkers include single amino acids, peptides, polypeptides, and linkers lacking any of the aforementioned such as various non-proteinaceous carbon chains, whether branched or cyclic (see e.g. Alley S et al., Bioconjug Chem 19: 759-65 (2008) Ducry L, Stump B, Bioconjug Chem 21: 5-13 (2010); Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)).
[0324] Suitable linkers may be proteinaceous and comprise one or more amino acids, peptides, and/or polypeptides. Proteinaceous linkers are suitable for both recombinant fusion proteins and chemically linked conjugates. A proteinaceous linker typically has from about 2 to about 50 amino acid residues, such as, e.g., from about 5 to about 30 or from about 6 to about 25 amino acid residues. The length of the linker selected will depend upon a variety of factors, such as, e.g., the desired property or properties for which the linker is being selected (see e.g. Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)).
[0325] Suitable linkers may be non-proteinaceous, such as, e.g. chemical linkers (see e.g. Dosio F et al., Toxins 3: 848-83 (2011); Feld J et al., Oncotarget 4: 397-412 (2013)). Various non-proteinaceous linkers known in the art may be used to link cell-targeting moieties to toxin effector polypeptide and other components to form a cell-targeting molecule of the present invention, such as linkers commonly used to conjugate immunoglobulin-derived polypeptides to heterologous polypeptides. For example, polypeptide regions of the cell-targeting molecules of the present invention may be linked using the functional side chains of their amino acid residues and carbohydrate moieties such as, e.g., a carboxy, amine, sulfhydryl, carboxylic acid, carbonyl, hydroxyl, and/or cyclic ring groups. For example, disulfide bonds and thioether bonds may be used to link two or more polypeptides (see e.g. Fitzgerald D et al., Bioconjugate Chem 1: 264-8 (1990); Pasqualucci L et al., Haematologica 80: 546-56 (1995)). In addition, non-natural amino acid residues may be used with other functional side chains, such as ketone groups (see e.g. Axup J et al., Proc Natl Acad Sci U.S.A. 109: 16101-6 (2012); Sun S et al., Chembiochem Jul. 18 (2014); Tian F et al., Proc Natl Acad Sci USA 111: 1766-71 (2014)). Examples of non-proteinaceous chemical linkers include but are not limited to N-succinimidyl (4-iodoacetyl)-aminobenzoate, S(N-succinimidyl) thioacetate (SATA), N-succinimidyl-oxycarbonyl-cu-methyl-a-(2-pyridyldithio) toluene (SMPT), N-succinimidyl 4-(2-pyridyldithio)-pentanoate (SPP), succinimidyl 4-(N-maleimidomethyl) cyclohexane carboxylate (SMCC or MCC), sulfosuccinimidyl (4-iodoacetyl)-aminobenzoate, 4-succinimidyl-oxycarbonyl--(2-pyridyldithio) toluene, sulfosuccinimidyl-6-(-methyl--(pyridyldithiol)-toluamido) hexanoate, N-succinimidyl-3-(2-pyridyldithio)-proprionate (SPDP), succinimidyl 6(3(-(2-pyridyldithio)-proprionamido) hexanoate, sulfosuccinimidyl 6(3(-(2-pyridyldithio)-propionamido) hexanoate, maleimidocaproyl (MC), maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-vc-PAB), 3-maleimidobenzoic acid N-hydroxysuccinimide ester (MBS), alpha-alkyl derivatives, sulfoNHS-ATMBA (sulfosuccinimidyl N-[3-(acetylthio)-3-methylbutyryl-beta-alanine]), sulfodicholorphenol, 2-iminothiolane, 3-(2-pyridyldithio)-propionyl hydrazide, Ellman's reagent, dichlorotriazinic acid, and S-(2-thiopyridyl)-L-cysteine (see e.g. Thorpe P et al., Eur J Biochem 147: 197-206 (1985); Thorpe P et al., Cancer Res 47: 5924-31 (1987); Thorpe P et al., Cancer Res 48: 6396-403 (1988); Grossbard M et al., Blood 79: 576-85 (1992); Lui C et al., Proc Natl Acad Sci USA 93: 8618-23 (1996); Doronina S et al., Nat Biotechnol 21: 778-84 (2003); Feld J et al., Oncotarget 4: 397-412 (2013)).
[0326] Suitable linkers, whether proteinaceous or non-proteinaceous, may include, e.g., protease sensitive, environmental redox potential sensitive, pH sensitive, acid cleavable, photocleavable, and/or heat sensitive linkers (see e.g. Dosio F et al., Toxins 3: 848-83 (2011); Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013); Feld J et al., Oncotarget 4: 397-412 (2013)).
[0327] Proteinaceous linkers may be chosen for incorporation into recombinant, fusion protein, cell-targeting molecules of the present invention. For recombinant fusion cell-targeting proteins of the invention, linkers typically comprise about 2 to 50 amino acid residues, preferably about 5 to 30 amino acid residues (Argos P, J Mol Biol 211: 943-58 (1990); Williamson M, Biochem 297: 240-60 (1994); George R, Heringa J, Protein Eng 15: 871-9 (2002); Kreitman R, AAPS J 8: E532-51 (2006)). Commonly, proteinaceous linkers comprise a majority of amino acid residues with polar, uncharged, and/or charged residues, such as, e.g., threonine, proline, glutamine, glycine, and alanine (see e.g. Huston J et al. Proc Natl Acad Sci U.S.A. 85: 5879-83 (1988); Pastan I et al., Annu Rev Med 58: 221-37 (2007); Li J et al., Cell Immunol 118: 85-99 (1989); Cumber A et al. Bioconj Chem 3: 397-401 (1992); Friedman P et al., Cancer Res 53: 334-9 (1993); Whitlow M et al., Protein Engineering 6: 989-95 (1993); Siegall C et al., J Immunol 152: 2377-84 (1994); Newton et al. Biochemistry 35: 545-53 (1996); Ladurner et al. J Mol Biol 273: 330-7 (1997); Kreitman R et al., Leuk Lymphoma 52: 82-6 (2011); U.S. Pat. No. 4,894,443). Non-limiting examples of proteinaceous linkers include alanine-serine-glycine-glycine-proline-glutamate (ASGGPE) (SEQ ID NO:1190), valine-methionine (VM), alanine-methionine (AM), AM(G.sub.2 to 4S).sub.xAM (SEQ ID NO:1191) where G is glycine, S is serine, and x is an integer from 1 to 10.
[0328] Proteinaceous linkers may be selected based upon the properties desired. Proteinaceous linkers may be chosen by the skilled worker with specific features in mind, such as to optimize one or more of the fusion protein's folding, stability, expression, solubility, pharmacokinetic properties, pharmacodynamic properties, and/or the activity of the fused domains in the context of a fusion construct as compared to the activity of the same domain by itself. For example, proteinaceous linkers may be selected based on flexibility, rigidity, and/or cleavability (see e.g. Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)). The skilled worker may use databases and linker design software tools when choosing linkers. Certain linkers may be chosen to optimize expression (see e.g. Turner D et al., J Immunol Methods 205: 43-54 (1997)). Certain linkers may be chosen to promote intermolecular interactions between identical polypeptides or proteins to form homomultimers or different polypeptides or proteins to form heteromultimers. For example, proteinaceous linkers may be selected which allow for desired non-covalent interactions between polypeptide components of the cell-targeting proteins of the invention, such as, e.g., interactions related to the formation dimers and other higher order multimers (see e.g. U.S. Pat. No. 4,946,778).
[0329] Flexible proteinaceous linkers are often greater than 12 amino acid residues long and rich in small, non-polar amino acid residues, polar amino acid residues, and/or hydrophilic amino acid residues, such as, e.g., glycines, serines, and threonines (see e.g. Bird R et al., Science 242: 423-6 (1988); Friedman P et al., Cancer Res 53: 334-9 (1993); Siegall C et al., J Immunol 152: 2377-84 (1994)). Flexible proteinaceous linkers may be chosen to increase the spatial separation between components and/or to allow for intramolecular interactions between components. For example, various GS linkers are known to the skilled worker and are composed of multiple glycines and/or one or more serines, sometimes in repeating units, such as, e.g., (G.sub.xS).sub.n (SEQ ID NO: 1192), (S.sub.xG).sub.n (SEQ ID NO: 1193), (GGGGS).sub.n (SEQ ID NO: 1194), and (G).sub.n (SEQ ID NO: 1195). in which x is 1 to 6 and n is 1 to 30 (see e.g. WO 96/06641). Non-limiting examples of flexible proteinaceous linkers include GKSSGSGSESKS (SEQ ID NO: 1196), GSTSGSGKSSEGKG (SEQ ID NO: 1197), GSTSGSGKSSEGSGSTKG (SEQ ID NO: 1198), GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1199), EGKSSGSGSESKEF (SEQ ID NO: 1200), SRSSG (SEQ ID NO: 1201), and SGSSC (SEQ ID NO: 1202).
[0330] Rigid proteinaceous linkers are often stiff alpha-helical structures and rich in proline residues and/or one or more strategically placed prolines (see Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)). Rigid linkers may be chosen to prevent intramolecular interactions between linked components.
[0331] Suitable linkers may be chosen to allow for in vivo separation of components, such as, e.g., due to cleavage and/or environment-specific instability (see Dosio F et al., Toxins 3: 848-83 (2011); Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)). The skilled worker knows how to make and use linkers designed to be stable, particularly in extracellular environments like in the blood plasma of a chordate's circulatory system, but to be cleaved in certain intracellular environments having unique characteristics, such as, e.g., in certain cell-types and/or intracellular compartments due to the presence of certain proteolytic activities, redox environments, and/or pH environments, thereby separating linked components of the molecule of the present invention.
[0332] In vivo cleavable proteinaceous linkers are capable of unlinking by proteolytic processing and/or reducing environments often at a specific site within an organism or inside a certain cell type (see e.g. Doronina S et al., Bioconjug Chem 17: 144-24 (2006); Erickson H et al., Cancer Res 66: 4426-33 (2006)). In vivo cleavable proteinaceous linkers often comprise protease sensitive motifs and/or disulfide bonds formed by one or more cysteine pairs (see e.g. Pietersz G et al., Cancer Res 48: 4469-76 (1998); The J et al., J Immunol Methods 110: 101-9 (1998); see Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)). In vivo cleavable proteinaceous linkers may be designed to be sensitive to proteases that exist only at certain locations in an organism, compartments within a cell, and/or become active only under certain physiological or pathological conditions (such as, e.g., proteases with abnormally high levels, proteases overexpressed at certain disease sites, and proteases specifically expressed by a pathogenic microorganism). For example, there are proteinaceous linkers known in the art which are cleaved by proteases present only intracellularly, proteases present only within specific cell types, and proteases present only under pathological conditions like cancer or inflammation, such as, e.g., R-x-x-R motif and AMGRSGGGCAGNRVGSSLSCGGLNLQAM (SEQ ID NO:1203).
[0333] In certain embodiments of the cell-targeting molecules of the present invention, a linker may be used which comprises one or more protease sensitive sites to provide for cleavage by a protease present within a target cell. In certain embodiments of the cell-targeting molecules of the invention, a linker may be used which is not cleavable to reduce unwanted toxicity after administration to a vertebrate organism (see e.g. Polson et al., Cancer Res 69: 2358-(2009)).
[0334] Suitable linkers may include, e.g., protease sensitive, environmental redox potential sensitive, pH sensitive, acid cleavable, photocleavable, and/or heat sensitive linkers, whether proteinaceous or non-proteinaceous (see Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013)).
[0335] Suitable cleavable linkers may include linkers comprising cleavable groups which are known in the art such as, e.g., linkers noted by Zarling D et al., J Immunol 124: 913-20 (1980); Jung S, Moroi M, Biochem Biophys Acta 761: 152-62 (1983); Bouizar Z et al., Eur J Biochem 155: 141-7 (1986); Park L et al., J Biol Chem 261: 205-10 (1986); Browning J, Ribolini A, J Immunol 143: 1859-67 (1989); Joshi S, Burrows R, J Biol Chem 265: 14518-25 (1990); Choi W et al., J Bioact Compat Polym 14: 447-56 (1999); Jensen K et al., Bioconjug Chem 13: 975-84 (2002); Christie R, Grainger, D, Adv Drug Deliv Rev 55: 421-37 (2003)).
[0336] Suitable linkers may include pH sensitive linkers. For example, certain suitable linkers may be chosen for their instability in lower pH environments to provide for dissociation inside a subcellular compartment of a target cell. For example, linkers that comprise one or more trityl groups, derivatized trityl groups, bismaleimidoethoxy propane groups, adipic acid dihydrazide groups, and/or acid labile transferrin groups, may provide for release of components of the cell-targeting molecules of the invention, e.g. a polypeptide component, in environments with specific pH ranges (see e.g. Welhner H et al., J Biol Chem 266: 4309-14 (1991); Fattom A et al., Infect Immun 60: 584-9 (1992)). Certain linkers may be chosen which are cleaved in pH ranges corresponding to physiological pH differences between tissues, such as, e.g., the pH of tumor tissue is lower than in healthy tissues (see e.g. U.S. Pat. No. 5,612,474).
[0337] Photocleavable linkers are linkers that are cleaved upon exposure to electromagnetic radiation of certain wavelength ranges, such as light in the visible range (see e.g. Goldmacher V et al., Bioconj Chem 3: 104-7 (1992)). Photocleavable linkers may be used to release a component of a cell-targeting molecule of the invention, e.g. a polypeptide component, upon exposure to light of certain wavelengths. Non-limiting examples of photocleavable linkers include a nitrobenzyl group as a photocleavable protective group for cysteine, nitrobenzyloxycarbonyl chloride cross-linkers, hydroxypropylmethacrylamide copolymer, glycine copolymer, fluorescein copolymer, and methylrhodamine copolymer (Hazum E et al., Pept Proc Eur Pept Symp, 16th, Brunfeldt K, ed., 105-110 (1981); Senter et al., Photochem Photobiol 42: 231-7 (1985); Yen et al., Makromol Chem 190: 69-82 (1989); Goldmacher V et al., Bioconj Chem 3: 104-7 (1992)). Photocleavable linkers may have particular uses in linking components to form cell-targeting molecules of the invention designed for treating diseases, disorders, and conditions that can be exposed to light using fiber optics.
[0338] In certain embodiments of the cell-targeting molecules of the present invention, a cell-targeting binding region is linked to a toxin effector polypeptide using any number of means known to the skilled worker, including both covalent and noncovalent linkages (see e.g. Chen X et al., Adv Drug Deliv Rev 65: 1357-69 (2013); Behrens C, Liu B, MAbs 6: 46-53 (2014)).
[0339] In certain embodiments of the cell-targeting molecules of the present invention, the protein comprises a binding region which is a scFv with a linker connecting a heavy chain variable (V.sub.H) domain and a light chain variable (V.sub.L) domain. There are numerous linkers known in the art suitable for this purpose, such as, e.g., the 15-residue (Gly4Ser).sub.3 peptide (SEQ ID NO:1204). Suitable scFv linkers which may be used in forming non-covalent multivalent structures include GGS, GGGS (Gly3Ser or G3 S) (SEQ ID NO:1205), GGGGS (Gly4Ser or G45) (SEQ ID NO:1206), GGGGSGGG (SEQ ID NO:1207), GGSGGGG (SEQ ID NO:1208), GSTSGGGSGGGSGGGGSS (SEQ ID NO:1209), and GSTSGSGKPGSSEGSTKG (SEQ ID NO:1210) (Phckthun A, Pack P, Immunotechnology 3: 83-105 (1997); Atwell J et al., Protein Eng 12: 597-604 (1999); Wu A et al., Protein Eng 14: 1025-33 (2001); Yazaki P et al., J Immunol Methods 253: 195-208 (2001); Carmichael J et al., J Mol Biol 326: 341-51 (2003); Arndt M et al., FEBS Lett 578: 257-61 (2004); Bie C et al., World J Hepatol 2: 185-91 (2010)).
[0340] In certain embodiments of the cell-targeting molecule of the present invention, the molecule comprises a linker between the Shiga toxin effector polypeptide and the binding region wherein the linker has one or more cysteine residues. In certain further embodiments, there is a molecule covalently linked to the cysteine residue via its sulfhydryl functional group.
[0341] In certain embodiments of the cell-targeting molecules of the present invention, the cell-targeting molecule comprises the linker comprising or consisting essentially of any one of SEQ ID NOs: 757-761 and 768-772.
[0342] In certain embodiments of the cell-targeting molecule of the present invention, the molecule comprises the linker comprising the polypeptide GGGC (SEQ ID NO:1211), such as, e.g., the linker comprising or consisting essentially of GGGGCGG (SEQ ID NO:1212), GGGGSGGGGCGG (SEQ ID NO:1213), GGGGCGGGGSGG (SEQ ID NO:1214), GGGGSGGGGSGGGGC (SEQ ID NO:1215), GGGGSGGGGCGGGGS (SEQ ID NO:1216), GGGGCGGGGCSGGGS (SEQ ID NO:1217), GGGGSGGGGCGGGGSSGGGGSSGGGGS (SEQ ID NO:1218), GGGGCGGGGSGGGGSSGGGGSSGGGGS (SEQ ID NO:1219), GGGGSGGGGSGGGGCSGGGGSSGGGGS (SEQ ID NO:1220), GGGGSGGGGSGGGGSCGGGGSSGGGGS (SEQ ID NO:1221), GGGGSGGGGSGGGGSSGGGGCSGGGGS (SEQ ID NO:1222), GGGGSGGGGSGGGGSSGGGGSCGGGGS (SEQ ID NO:1223), or GGGGSGGGGSGGGGSGGGGSSGGGGC (SEQ ID NO:1224).
[0343] In certain embodiments of the cell-targeting molecule of the present invention, the molecule comprises the linker comprising the polypeptide GGGK (SEQ ID NO:1225), such as, e.g., the linker comprising or consisting essentially of GGGGKGG (SEQ ID NO:1226), GGGGSGGGGKGG (SEQ ID NO:1227), GGGGKGGGGSGG (SEQ ID NO:1228), GGGGSGGGGSGGGGK (SEQ ID NO:1229), GGGGSGGGGKGGGGS (SEQ ID NO:1230), GGGGKGGGGCSGGGS (SEQ ID NO:1231), GGGGCGGGGKSGGGS (SEQ ID NO:1232), GGGGSGGGGKGGGGSSGGGGSSGGGGS (SEQ ID NO:1233), GGGGKGGGGSGGGGSSGGGGSSGGGGS (SEQ ID NO:1234), GGGGSGGGGSGGGGKSGGGGSSGGGGS (SEQ ID NO:1235), GGGGSGGGGSGGGGSKGGGGSSGGGGS (SEQ ID NO:1236), GGGGSGGGGSGGGGSSGGGGKSGGGGS (SEQ ID NO:1237), GGGGSGGGGSGGGGSSGGGGSKGGGGS (SEQ ID NO:1238), or GGGGSGGGGSGGGGSGGGGSSGGGGK (SEQ ID NO:1239).
[0344] Suitable methods for linkage of the components of the cell-targeting molecules of the present invention may be by any method presently known in the art for accomplishing such, as long as the attachment does not substantially impede the binding capability of the cell-targeting agent or binding region, the cellular internalization of the cell-targeting molecule, the intracellular delivery of a cargo to a subcellular compartment or specific location, and/or the subcellular routing of the toxin effector polypeptide, each of which can be determined by an appropriate assay, including by assays described herein.
F. Cargos, Heterologous Matter, Conjugated Moieties, Cell-Targeting Molecule Altering Agents, and Additional Exogenous Materials
[0345] In certain embodiments, a molecule of the present invention comprises matter heterologous to Shiga toxins, such as, e.g., a cargo, conjugated molecule, additional exogenous material, and/or cell-targeting molecule altering agent. A molecule conjugated to a cell-targeting molecule represents a conjugated moiety. Such a heterologous matter (e.g. an atom or molecule) linked to a Shiga toxin effector polypeptide may be a matter which is foreign to the target cell and/or is not present in intended target cells in desirable amounts. In certain embodiments, the conjugated matter is an atomic or molecular cargo, heterologous molecule, additional exogenous material, and/or cell-targeting molecule altering agent, such as, e.g., a CD8+ T-cell epitope and/or antigen, radionucleide, peptide, detection-promoting agent, protein, small molecule chemotherapeutic agent, and/or polynucleotide.
[0346] In certain embodiments, the conjugated matter is or comprises an atom, such as a radionucleide. In certain embodiments, the radionucleide is .sup.211At, .sup.131I, .sup.125I, .sup.90Y, .sup.111In, .sup.186Re, .sup.188Re, .sup.153Sm, .sup.212Bi, .sup.32P, .sup.60C, and/or a radioactive isotope of lutetium.
[0347] In certain embodiments, the conjugated matter is a lipid, serum albumin binding molecule, antibiotic, cytotoxic agent, detection-promoting agent, peptide, protein, enzyme, nucleic acid, and/or protein-nucleic acid complex (see e.g. Liu B, Brief Funct Genomic Proteomic 6: 112-9 (2007); Endoh T, Ohtsuki T, Adv Drug Deliv Rev 61: 704-9 (2009)). In certain embodiments, the cell-targeting molecules of the present invention comprise a conjugated matter which is a molecule altering agent meant to function prior to or during target cell internalization of the cell-targeting molecule. In certain other embodiments, the cell-targeting molecules of the present invention comprise a conjugated matter which is a cargo (e.g. an additional exogenous material) meant to function after target cell internalization of the cell-targeting molecule. In certain embodiments, the cell-targeting molecules of the present invention comprise a conjugated matter which is an additional exogenous material or cargo, such as, e.g., a polypeptide comprising a pro-apoptotic effector like, e.g., fragments of caspase-3, caspase-6, granzyme B, tBid, and apoptosis inducing factor (AIF) (see e.g., Jia 1, et al., Cancer Res 63: 3257-62 (2003); Xu Y et al., J Immunol 173: 61-7 (2004); Wang T et al., Cancer Res 67: 11830-9 (2007); Shan L et. al., Cancer Biol Ther 11: 1717-22 (2008); Qiu X et al., Mol Cancer Ther 7: 1890-9 (2008)), In certain embodiments, the conjugated matter is a peptide comprising or consisting essentially of a CD8+ T-cell epitope and/or antigen.
[0348] Non-limiting examples of cell-targeting molecule altering agents include various polyethylene glycol molecules, lipids, and liposomes because these agents can alter, e.g., the immunogenicity and/or pharmacokinetics of the cell-targeting molecule.
[0349] In certain embodiments, the conjugated matter is a non-proteinaceous polymer, e.g., a polyethylene glycol, polypropylene glycol, or polyoxyalkylene (see e.g., U.S. Pat. Nos. 4,179,337; 4,301,144; 4,640,835; 4,670,417; 4,791,192; and 4,496,689). Any one of these can function as a cell-targeting molecule altering agent.
[0350] In certain embodiments, the cell-targeting molecule of the present invention comprises a cargo molecule covalently conjugated to an amino acid residue in a proteinaceous component of the cell-targeting molecule. In certain further embodiments, the cell-targeting molecule of the present invention comprises a cargo molecule covalently conjugated to a cysteine, lysine, or histidine residue in the cell-targeting molecule. In certain further embodiments, the cell-targeting molecule of the present invention comprises a cargo molecule covalently conjugated to a Shiga toxin effector polypeptide component of the cell-targeting molecule via a cysteine, lysine, or histidine residue in the Shiga toxin effector polypeptide. In certain further embodiments, the cargo is covalently linked to the cysteine residue via its sulfhydryl functional group. In certain further embodiments, the cargo molecule is released from the cell-targeting molecule upon reaching the endoplasmic reticulum or another compartment having a reducing environment due to the reduction of a disulfide bond between the cargo and the Shiga toxin effector polypeptide (see e.g. El Alaoui A et al., Angew Chem Int Ed Engl 46: 6469-72 (2007)).
[0351] In certain embodiments, the cell-targeting molecule of the present invention comprises a nucleic acid cargo conjugated to a proteinaceous component of the cell-targeting molecule via a disulfide bond (see e.g. Muratovska A, Eccles M, FEBS Lett 558: 63-8 (2004); Ishihara T et al., Drug Deliv 16: 153-9 (2009)).
[0352] In certain embodiments, the cargo is a protein or polypeptide comprising an enzyme. In certain other embodiments, the cargo is a nucleic acid, such as, e.g. a ribonucleic acid that functions as a small inhibiting RNA (siRNA) or microRNA (miRNA). In certain embodiments, the cargo is an antigen, such as antigens derived from pathogens, bacterial proteins, viral proteins, proteins mutated in cancer, proteins aberrantly expressed in cancer, or T-cell complementary determining regions. For example, cargos may include antigens, such as those antigens presented by antigen-presenting cells infected by bacteria, and T-cell complementary determining regions capable of functioning as exogenous antigens. Cargo molecules comprising polypeptides or proteins may optionally comprise one or more antigens whether known or unknown to the skilled worker.
[0353] In certain embodiments of the cell-targeting molecules of the present invention, all heterologous antigens and/or epitopes associated with the Shiga toxin effector polypeptide are arranged in the cell-targeting molecule amino-terminal to the carboxy-terminus of the Shiga toxin A1 fragment region of the Shiga toxin effector polypeptide. In certain further embodiments, all heterologous antigens and/or epitopes associated with the Shiga toxin effector polypeptide are associated, either directly or indirectly, with the Shiga toxin effector polypeptide at a position amino-terminal to the carboxy-terminus of the Shiga toxin A1 fragment region of the Shiga toxin effector polypeptide. In certain further embodiments, all additional exogenous material(s) which is an antigen is arranged amino-terminal to the Shiga toxin effector polypeptide, such as, e.g., fused directly or indirectly to the amino terminus of the Shiga toxin effector polypeptide.
[0354] In certain embodiments, the cell-targeting molecule of the present invention comprises a protein-nucleic acid complex cargo conjugated to a proteinaceous component of the cell-targeting molecule via a disulfide bond.
[0355] In certain embodiments, the cargo and/or detection-promoting agent is a fluorophore, such as, e.g., a maleimide derivative of an Alexa Fluor fluorophore for conjugating to a thiol functional group of an amino acid residue of a molecule of the present invention.
[0356] In certain embodiments, the cell-targeting molecule of the present invention comprises a zymoxin, which is an inactive viral enzyme that may be activated by proteolytic cleavage (see e.g. Shapira A et al., PLoS One 6: e15916 (2011)).
[0357] In certain embodiments, the cargo is a proapoptotic peptide, polypeptide, or protein, such as, e.g., BCL-2, caspases (e.g. fragments of caspase-3 or caspase-6), cytochromes, granzyme B, apoptosis-inducing factor (AIF), BAX, tBid (truncated Bid), and proapoptotic fragments or derivatives thereof (see e.g., Ellerby H et al., Nat Med 5: 1032-8 (1999); Mai J et al., Cancer Res 61: 7709-12 (2001); Jia et al., Cancer Res 63: 3257-62 (2003); Liu Y et al., Mol Cancer Ther 2: 1341-50 (2003); Perea S et al., Cancer Res 64: 7127-9 (2004); Xu Y et al., J Immunol 173: 61-7 (2004); Dlken B et al., Cell Death Differ 13: 576-85 (2006); Wang T et al., Cancer Res 67: 11830-9 (2007); Kwon M et al., Mol Cancer Ther 7: 1514-22 (2008); Qiu X et al., Mol Cancer Ther 7: 1890-9 (2008); Shan L et al., Cancer Biol Ther 1717-22 (2008); Wang F et al., Clin Cancer Res 16: 2284-94 (2010); Kim J et al., J Virol 85: 1507-16 (2011)).
[0358] In certain embodiments, the cargo is a cytotoxic agent, such as, e.g., a small molecule chemotherapeutic agent, anti-neoplastic agent, cytotoxic antibiotic, alkylating agent, antimetabolite, topoisomerase inhibitor, and/or tubulin inhibitor. Non-limiting examples of cytotoxic agents suitable for use with the present invention include aziridines, cisplatins, tetrazines, procarbazine, hexamethylmelamine, vinca alkaloids, taxanes, camptothecins, etoposide, doxorubicin, mitoxantrone, teniposide, novobiocin, aclarubicin, anthracyclines, actinomycin, amanitin, amatoxins, bleomycin, centanamycin (indolecarboxamide), plicamycin, mitomycin, daunorubicin, epirubicin, idarubicins, dolastatins, maytansines, maytansionoids, duromycin, docetaxel, duocarmycins, adriamycin, calicheamicin, auristatins, pyrrolobenzodiazepines, pyrrolobenzodiazepine dimers (PBDs), carboplatin, 5-fluorouracil (5-FU), capecitabine, mitomycin C, paclitaxel, 1,3-Bis(2-chloroethyl)-1-nitrosourea (BCNU), rifampicin, cisplatin, methotrexate, gemcitabine, aceglatone, acetogenins (e.g. bullatacin and bullatacinone), aclacinomysins, AG1478, AG1571, aldophosphamide glycoside, alkyl sulfonates (e.g., busulfan, improsulfan, and piposulfan), alkylating agents (e.g. thiotepa and cyclosphosphamide), aminolevulinic acid, aminopterin, amsacrine, ancitabine, anthramycin, arabinoside, azacitidine, azaserine, aziridines (e.g., benzodopa, carboquone, meturedopa, and uredopa), azauridine, bestrabucil, bisantrene, bisphosphonates (e.g. clodronate), bleomycins, bortezomib, bryostatin, cactinomycin, callystatin, carabicin, carminomycin, carmofur, carmustine, carzinophilin, CC-1065, chlorambucil, chloranbucil, chlornaphazine, chlorozotocin, chromomycinis, chromoprotein enediyne antibiotic chromophores, CPT-11, cryptophycins (e.g. cryptophycin 1 and cryptophycin 8), cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunomycin, defofamine, demecolcine, detorubicin, diaziquone, 6-diazo-5-oxo-L-norleucine, dideoxyuridine, difluoromethylornithine (DMFO), doxifluridine, doxorubicins (e.g., morpholinodoxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolinodoxorubicin, and deoxydoxorubicin), dynemicins, edatraxate, edatrexate, eleutherobins, elformithine, elliptinium acetate, enediyne antibiotics (e.g. calicheamicins), eniluracil, enocitabine, epirubicins, epothilone, esorubicins, esperamicins, estramustine, ethylenimines, 2-ethylhydrazide, etoglucid, fludarabine, folic acid analogues (e.g., denopterin, methotrexate, pteropterin, and trimetrexate), folic acid replenishers (e.g. frolinic acid), fotemustine, fulvestrant, gacytosine, gallium nitrate, gefitinib, gemcitabine, hydroxyurea, ibandronate, ifosfamide, imatinib mesylate, erlotinib, fulvestrant, letrozole, PTK787/ZK 222584 (Novartis, Basel, CH), oxaliplatin, leucovorin, rapamycin, lapatinib, lonafarnib, sorafenib, methylamelamines (e.g., altretamine, triethy lenemelamine, triethy lenephosphoramide, triethylenethiophosphoramide and trimethylomelamine), pancratistatins, sarcodictyins, spongistatins, nitrogen mustards (e.g., chlorambucil, chlornaphazine, cyclophosphamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, and uracil mustard), nitrosureas (e.g., carmustine, fotemustine, lomustine, nimustine, and ranimnustine), dynemicins, neocarzinostatin chromophores, anthramycin, detorubicin, epirubicins, marcellomycins, mitomycins (e.g. mitomycin C), mycophenolic acid, nogalamycins, olivomycins, peplomycins, potfiromycins, puromycins, quelamycins, rodorubicins, ubenimex, zinostatins, zorubicins, purine analogs (e.g., fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine), pyrimidine analogs (e.g., ancitabine, azacitidine, 6-azauridine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine), aceglatone, lentinan, lonidainine, maytansinoids (e.g. maytansins and ansamitocins), mitoguazone, mitoxantrone, mopidanmol, nitraerine, pentostatin, phenamet, pirarubicin, podophyllinic acid, 2-ethylhydrazide, rhizoxin, sizofuran, spirogermanium, tenuazonic acid, triaziquone, 2,2,2trichlorotriethylamine, trichothecenes (e.g., T-2 toxin, verracurin A, roridin A, and anguidine), urethan, vindesine, mannomustine, mitobronitol, mitolactol, pipobroman, arabinoside, cyclophosphamide, toxoids (e.g. paclitaxel and doxetaxel), 6-thioguanine, mercaptopurine, platinum, platinum analogs (e.g. cisplatin and carboplatin), etoposide (VP-16), mitoxantrone, vinorelbine, novantrone, daunomycin, xeloda, topoisomerase inhibitor RFS 2000, retinoids (e.g. retinoic acid), capecitabine, lomustine, losoxantrone, mercaptopurines, nimustine, nitraerine, rapamycin, razoxane, roridin A, spongistatins, streptonigrins, streptozocins, sutent, T-2 toxin, thiamiprine, thiotepa, toxoids (e.g. paclitaxel and doxetaxel), tubercidins, verracurin A, vinblastine, vincristine, and structural analogs of any of the aforementioned (e.g. synthetic analogs), and/or derivatives of any of the aforementioned (see e.g., Lindell T et al., Science 170: 447-9 (1970); Remillard S et al., Science 189: 1002-5 (1975); Ravry M et al., Am J Clin Oncol 8: 148-50 (1985); Ravry M et al., Cancer Treat Rep 69: 1457-8 (1985); Sternberg C et al., Cancer 64: 2448-58 (1989); Bai R et al., Biochem Pharmacol 39: 1941-9 (1990); Boger D, Johnson D, Proc Natl Acad Sci USA 92: 3642-9 (1995); Beck J et al., Leuk Lymphoma 41: 117-24 (2001); Cassady J et al., Chem Pharm Bull (Tokyo) 52: 1-26 (2004); Sapra P et al., Clin Cancer Res 11: 5257-64 (2005); Okeley N et al., Clinc Cancer Res 16: 888-97 (2010); Oroudjev E et al., Mol Cancer Ther 9: 2700-13 (2010); Ellestad G, Chirality 23: 660-71 (2011); Kantarjian H et al., Lancet Oncol 13: 403-11 (2012); Moldenhauer G et al., J Natl Cancer Inst 104: 622-34 (2012); Gromek S, Balunas M, Curr Top Med Chem 14: 2822-34 (2015); Meulendijks D et al., Invest New Drugs 34: 119-28 (2016)).
[0359] In certain embodiments of the cell-targeting molecules of the present invention, the additional exogenous material is a polypeptide characterized as highly efficient in delivering various molecules into cells, such polypeptides also known as cell penetrating peptides (CPPs) or protein transduction domains (PTDs) (see Futaki S et al., Biochemistry 41: 7925-30 (2002); Wender P et al., J Am Chem Soc 124: 13382-3 (2002); Dietz G, Bahr M, Mol Cell Neurosci 27: 85-131 (2004); Vives E, J Control Release 109: 77-85 (2005); Vive{grave over (s)} E et al., Biochim Biophys Acta 17866: 126-38 (2008); van den Berg A, Dowdy S, Curr Opin Biotechnol 22: 888-93 (2011); Copolovici D et al., ACS Nano 8: 1972-94 (2014); Kauffman W et al., Trends Biochem Sci 40: 749-64 (2015); WO 2003106491; U.S. Pat. Nos. 7,579,318; 7,943,581; 8,242,081). In certain embodiments, the additional exogenous material comprises both a CPP and/or PTD and a nucleic acid and optionally a cationic peptide (see e.g. Lehto T et al., Expert Opin Drug Deliv 9: 823-36 (2012); Shukla R et al., Mol Pharm 11: 3395-408 (2014); Beloor J et al., Ther Deliv 6: 491-507 (2015); Chuah J et al., Biomacromolecules 10.1021/acs.biomac.6b01056 (2016); Cerrato C et al., Expert Opin Drug Deliv 1-11 (2016); Tai W, Gao X, Adv Drug Deliv Rev pii: 50169-409X: 30236-8 (2016); Wada S et al., Bioorg Med Chem 15: 4478-85 (2016)). In certain embodiments, the additional exogenous material comprises a CPP and/or PTD for intracellular targeting of another additional exogenous material (see e.g. Sakhrani N, Padh H, Drug Des Devel Ther 7: 585-99 (2013); Li H et al., Int J Mol Sci 16: 19518-36 (2015); Cerrato C et al., Expert Opin Drug Deliv 1-11 (2016)).
II. Examples of Structural Variants of the Cell-Targeting Molecules of the Present Invention
[0360] In certain embodiments of the cell-targeting molecules of the present invention, many of the molecule's components have already been described, such as the binding region, linker, and/or toxin effector polypeptide (see e.g. WO 2005/092917, WO 2007/033497, US2009/0156417, JP4339511, EP1727827, DE602004027168, EP1945660, JP4934761, EP2228383, US2013/0196928, WO 2014/164680, WO 2014/164693, WO 2015/138435, WO 2015/138452, WO 2015/113005, WO 2015/113007, US20150259428, WO 2015/191764, WO 2016/126950).
[0361] The skilled worker will recognize that variations may be made to the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention, and polynucleotides encoding any of the former, without diminishing their biological activities, e.g., by maintaining the overall structure and function of the Shiga toxin effector polypeptide, such as in conjunction with one or more 1) endogenous epitope disruptions which reduce antigenic and/or immunogenic potential, 2) furin-cleavage motif disruptions which reduce proteolytic cleavage, and/or 3) embedded or inserted epitopes which reduce antigenic and/or immunogenic potential and/or are capable of being delivered to a MHC I molecule for presentation on a cell surface. For example, some modifications may facilitate expression, facilitate purification, improve pharmacokinetic properties, and/or improve immunogenicity. Such modifications are well known to the skilled worker and include, for example, a methionine added at the amino-terminus to provide an initiation site, additional amino acids placed on either terminus to create conveniently located restriction sites or termination codons, and biochemical affinity tags fused to either terminus to provide for convenient detection and/or purification. A common modification to improve the immunogenicity of a polypeptide produced using a non-chordate system (e.g. a prokaryotic cell) is to remove, after the production of the polypeptide, the starting methionine residue, which may be formylated during production, such as, e.g., in a bacterial host system, because, e.g., the presence of N-formylmethionine (fMet) might induce undesirable immune responses in chordates.
[0362] Also contemplated herein is the inclusion of additional amino acid residues at the amino and/or carboxy termini of a Shiga toxin effector polypeptide of the present invention, a cell-targeting molecule of the present invention, or a proteinaceous component of a cell-targeting molecules of the present invention, such as sequences for epitope tags or other moieties. The additional amino acid residues may be used for various purposes including, e.g., facilitating cloning, facilitating expression, post-translational modification, facilitating synthesis, purification, facilitating detection, and administration. Non-limiting examples of epitope tags and moieties are chitin binding protein domains, enteropeptidase cleavage sites, Factor Xa cleavage sites, FIAsH tags, FLAG tags, green fluorescent proteins (GFP), glutathione-S-transferase moieties, HA tags, maltose binding protein domains, myc tags, polyhistidine tags, ReAsH tags, strep-tags, strep-tag II, TEV protease sites, thioredoxin domains, thrombin cleavage site, and V5 epitope tags.
[0363] In certain of the above embodiments, the polypeptide sequence of the Shiga toxin effector polypeptides and/or cell-targeting molecules of the present invention are varied by one or more conservative amino acid substitutions introduced into the polypeptide region(s) as long as all required structural features are still present and the Shiga toxin effector polypeptide is capable of exhibiting any required function(s), either alone or as a component of a cell-targeting molecule. As used herein, the term conservative substitution denotes that one or more amino acids are replaced by another, biologically similar amino acid residue. Examples include substitution of amino acid residues with similar characteristics, e.g. small amino acids, acidic amino acids, polar amino acids, basic amino acids, hydrophobic amino acids and aromatic amino acids (see, for example, Table B). An example of a conservative substitution with a residue normally not found in endogenous, mammalian peptides and proteins is the conservative substitution of an arginine or lysine residue with, for example, ornithine, canavanine, aminoethylcysteine, or another basic amino acid. For further information concerning phenotypically silent substitutions in peptides and proteins see, e.g., Bowie J et al., Science 247: 1306-10 (1990).
TABLE-US-00002 TABLEB ExamplesofConservativeAminoAcid Substitutions I II III IV V VI VII VIII IX X XI XII XIII XIV A D H C F N A C F A C A A D G E K I W Q G M H C D C C E P Q R L Y S I P W F E D D G S N M T L Y G H G E K T V V H K N G P I N P H Q L Q S K R M R T N S R S V Q T T T R V S W P Y T
[0364] In the conservative substitution scheme in Table B, exemplary conservative substitutions of amino acids are grouped by physicochemical propertiesI: neutral, hydrophilic; II: acids and amides; III: basic; IV: hydrophobic; V: aromatic, bulky amino acids, VI hydrophilic uncharged, VII aliphatic uncharged, VIII non-polar uncharged, IX cycloalkenyl-associated, X hydrophobic, XI polar, XII small, XIII turn-permitting, and XIV flexible. For example, conservative amino acid substitutions include the following: 1) S may be substituted for C; 2) M or L may be substituted for F; 3) Y may be substituted for M; 4) Q or E may be substituted for K; 5) N or Q may be substituted for H; and 6) H may be substituted for N.
[0365] Additional conservative amino acid substitutions include the following: 1) S may be substituted for C; 2) M or L may be substituted for F; 3) Y may be substituted for M; 4) Q or E may be substituted for K; 5) N or Q may be substituted for H; and 6) H may be substituted for N.
[0366] In certain embodiments, the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention may comprise functional fragments or variants of a polypeptide region of the present invention described herein that have, at most, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions compared to a polypeptide sequence recited herein, as long as it (1) comprises at least one embedded or inserted, heterologous T-cell epitope and at least one amino acid is disrupted in an endogenous, B-cell and/or CD4+ T-cell epitope region provided in the Examples (see e.g. Tables 1-7 and/or 12), wherein the disrupted amino acid does not overlap with the embedded or inserted epitope; (2) comprises at least one embedded or inserted, heterologous T-cell epitope and a disrupted furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment derived region; or (3) comprises a disrupted furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment derived region and comprises at least one amino acid is disrupted in an endogenous, B-cell and/or CD4+ T-cell epitope region provided in the Examples (see e.g. Tables 1-7 and/or 12), wherein the disrupted amino acid does not overlap with the disrupted furin-cleavage motif. Variants of the Shiga toxin effector polypeptides and cell-targeting molecules of the invention are within the scope of the present invention as a result of changing a polypeptide described herein by altering one or more amino acid residues or deleting or inserting one or more amino acid residues, such as within the binding region or Shiga toxin effector polypeptide region, in order to achieve desired properties, such as changed cytotoxicity, changed cytostatic effects, changed immunogenicity, and/or changed serum half-life. The Shiga toxin effector polypeptides and cell-targeting molecules of the present invention may further be with or without a signal sequence.
[0367] Accordingly, in certain embodiments, the Shiga toxin effector polypeptides of the present invention comprise or consists essentially of amino acid sequences having at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99%, overall sequence identity to a naturally occurring Shiga toxin A Subunit or fragment thereof, such as, e.g., Shiga toxin A Subunit, such as SLT-1A (SEQ ID NO:1), StxA (SEQ ID NO:2), and/or SLT-2A (SEQ ID NO:3), wherein the Shiga toxin effector polypeptide (1) comprises at least one embedded or inserted, heterologous T-cell epitope and at least one amino acid is disrupted in an endogenous, B-cell and/or CD4+ T-cell epitope region provided in the Examples (see e.g. Tables 1-7 and/or 12), and wherein the disrupted amino acid does not overlap with the embedded or inserted epitope; (2) comprises at least one embedded or inserted, heterologous T-cell epitope and a disrupted furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment derived region; or (3) comprises a disrupted furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment derived region and comprises at least one amino acid is disrupted in an endogenous, B-cell and/or CD4+ T-cell epitope region provided in the Examples (see e.g. Tables 1-7 and/or 12), and wherein the disrupted amino acid does not overlap with the disrupted furin-cleavage motif.
[0368] In certain embodiments of the Shiga toxin effector polypeptides of the present invention, one or more amino acid residues may be mutated, inserted, or deleted in order to increase the enzymatic activity of the Shiga toxin effector polypeptide. In certain embodiments of the Shiga toxin effector polypeptides of the present invention, one or more amino acid residues may be mutated or deleted in order to reduce or eliminate catalytic and/or cytotoxic activity of the Shiga toxin effector polypeptide. For example, the catalytic and/or cytotoxic activity of the A Subunits of members of the Shiga toxin family may be diminished or eliminated by mutation or truncation.
[0369] The cytotoxicity of the A Subunits of members of the Shiga toxin family may be altered, reduced, or eliminated by mutation and/or truncation. The positions labeled tyrosine-77, glutamate-167, arginine-170, tyrosine-114, and tryptophan-203 have been shown to be important for the catalytic activity of Stx, Stx1, and Stx2 (Hovde C et al., Proc Natl Acad Sci USA 85: 2568-72 (1988); Deresiewicz R et al., Biochemistry 31: 3272-80 (1992); Deresiewicz R et al., Mol Gen Genet 241: 467-73 (1993); Ohmura M et al., Microb Pathog 15: 169-76 (1993); Cao C et al., Microbiol Immunol 38: 441-7 (1994); Suhan M, Hovde C, Infect Immun 66: 5252-9 (1998)). Mutating both glutamate-167 and arginine-170 eliminated the enzymatic activity of Slt-I A1 in a cell-free ribosome inactivation assay (LaPointe P et al., J Biol Chem 280: 23310-18 (2005)). In another approach using de novo expression of Slt-I A1 in the endoplasmic reticulum, mutating both glutamate-167 and arginine-170 eliminated Slt-I A1 fragment cytotoxicity at that expression level (LaPointe P et al., J Biol Chem 280: 23310-18 (2005)). A truncation analysis demonstrated that a fragment of StxA from residues 75 to 268 still retains significant enzymatic activity in vitro (Haddad J et al., J Bacteriol 175: 4970-8 (1993)). A truncated fragment of Slt-I A1 containing residues 1-239 displayed significant enzymatic activity in vitro and cytotoxicity by de novo expression in the cytosol (LaPointe P et al., J Biol Chem 280: 23310-18 (2005)). Expression of a Slt-I A1 fragment truncated to residues 1-239 in the endoplasmic reticulum was not cytotoxic because it could not retrotranslocate to the cytosol (LaPointe P et al., J Biol Chem 280: 23310-18 (2005)).
[0370] The most critical residues for enzymatic activity and/or cytotoxicity in the Shiga toxin A Subunits were mapped to the following residue-positions: asparagine-75, tyrosine-77, tyrosine-114, glutamate-167, arginine-170, arginine-176, and tryptophan-203 among others (Di R et al., Toxicon 57: 525-39 (2011)). In particular, a double-mutant construct of Stx2A containing glutamate-E167-to-lysine and arginine-176-to-lysine mutations was completely inactivated; whereas, many single mutations in Stx1 and Stx2 showed a 10-fold reduction in cytotoxicity. Further, truncation of Stx1A to 1-239 or 1-240 reduced its cytotoxicity, and similarly, truncation of Stx2A to a conserved hydrophobic residue reduced its cytotoxicity. The most critical residues for binding eukaryotic ribosomes and/or eukaryotic ribosome inhibition in the Shiga toxin A Subunit have been mapped to the following residue-positions arginine-172, arginine-176, arginine-179, arginine-188, tyrosine-189, valine-191, and leucine-233 among others (McCluskey A et al., PLoS One 7: e31191 (2012). However, certain modification may increase a Shiga toxin functional activity exhibited by a Shiga toxin effector polypeptide of the present invention. For example, mutating residue-position alanine-231 in Stx1A to glutamate increased Stx1A's enzymatic activity in vitro (Suhan M, Hovde C, Infect Immun 66: 5252-9 (1998)).
[0371] In certain embodiments of Shiga toxin effector polypeptides of the present invention derived from SLT-1A (SEQ ID NO:1) or StxA (SEQ ID NO:2), the one or more amino acid residues mutated include substitution of the asparagine at position 75, tyrosine at position 77, tyrosine at position 114, glutamate at position 167, arginine at position 170, arginine at position 176, and/or substitution of the tryptophan at position 203. Examples of such substitutions will be known to the skilled worker based on the prior art, such as asparagine at position 75 to alanine; tyrosine at position 77 to serine; substitution of the tyrosine at position 114 to serine; substitution of the glutamate position 167 to glutamate, glutamine, or lysine; substitution of the arginine at position 170 to alanine, glycine, or lysine; substitution of the arginine at position 176 to lysine; substitution of the tryptophan at position 203 to alanine; and/or substitution of the alanine at 231 with glutamate. Other mutations which either enhance or reduce Shiga toxin enzymatic activity and/or cytotoxicity are within the scope of the invention and may be determined using well-known techniques and assays disclosed herein.
[0372] In certain embodiments of the Shiga toxin effector polypeptide scaffolds of the present invention, the Shiga toxin effector polypeptide component of the scaffold is selected from any one of SEQ ID NOs: 233-756. In certain further embodiments of the Shiga toxin effector polypeptide scaffolds of the present invention, the Shiga toxin effector polypeptide component of the scaffold is selected from any one of SEQ ID NOs: 233-756 and the Shiga toxin effector polypeptide further comprises a linker. In certain further embodiments of the Shiga toxin effector polypeptide scaffolds of the present invention, the Shiga toxin effector polypeptide component of the scaffold is selected from any one of SEQ ID NOs: 233-756 and the Shiga toxin effector polypeptide scaffold further comprises a linker selected from any one of SEQ ID NOs: 757-761. In certain further embodiments of the Shiga toxin effector polypeptide scaffolds of the present invention, the Shiga toxin effector polypeptide component of the scaffold is selected from any one of SEQ ID NOs: 233-756 and the Shiga toxin effector polypeptide scaffold further comprises a linker selected from any one of SEQ ID NOs: 757-761 as long as there is a single, unique cysteine or lysine residue present in the scaffold outside of the Shiga toxin effector polypeptide. In certain further embodiments of the Shiga toxin effector polypeptide scaffolds of the present invention, the Shiga toxin effector polypeptide scaffold comprises or consists essentially of any one of SEQ ID NOs: 762-767.
[0373] In certain embodiments of the cell-targeting molecules of the present invention, the cell-targeting molecule comprises a Shiga toxin effector polypeptide component selected from any one of SEQ ID NOs: 5-756.
[0374] In certain embodiments of the cell-targeting molecules of the present invention, the cell-targeting molecule comprises a Shiga toxin effector polypeptide scaffold of the present invention. In certain further embodiments, the Shiga toxin effector polypeptide scaffold comprises only one cysteine and/or lysine residue. In certain further embodiments, the Shiga toxin effector polypeptide scaffold is selected from any one of SEQ ID NOs: 762-767.
[0375] In certain embodiments of the cell-targeting molecules of the present invention, the cell-targeting molecule comprises one or more of SEQ ID NOs: 757-761 and 768-772.
[0376] Certain embodiments of the cell-targeting molecules of the present invention comprise or consist essentially of any one of SEQ ID NOs: 773-829.
[0377] In certain embodiments of the cell-targeting molecules of the present invention, the cell-targeting molecule comprises a binding region comprising an immunoglobulin domain. In certain further embodiments of the cell-targeting molecule of the present invention, the binding region comprises a polypeptide(s) selected from the group consisting of: (a) a heavy chain variable (V.sub.H) domain comprising a HCDR1 comprising or consisting essentially of the amino acid sequences as shown in any one of SEQ ID NOs: 844, 850, 857, 863, 869, 875, 881, 885, 891, 897, 903, 909, 915, 921, 927, 933, 939, 948, 954, 960, 966, 972, 978, 984, 990, 996, 1002, 1008, 1014, 1020, 1026, 1032, 1035, 1041, 1044, 1050, 1056, 1062, 1065, 1071, 1077, 1083, 1089, and 1095, a HCDR2 comprising or consisting essentially of the amino acid sequences as shown in SEQ ID NOs: 845, 851, 856, 858, 864, 876, 886, 892, 898, 904, 910, 916, 922, 928, 934, 940, 949, 955, 961, 967, 973, 979, 985, 991, 997, 1003, 1009, 1015, 1021, 1027, 1036, 1042, 1045, 1051, 1057, 1063, 1066, 1072, 1078, 1084, 1090, and 1096, and a HCDR3 comprising or consisting essentially of the amino acid sequences as shown in any one of SEQ ID NOs: 846, 852, 859, 865, 870, 872, 877, 882, 887, 893, 899, 905, 911, 917, 923, 929, 935, 941, 950, 956, 962, 968, 974, 980, 982, 986, 992, 998, 1004, 1010, 1016, 1022, 1028, 1037, 1043, 1046, 1064, 1052, 1058, 1067, 1073, 1079, 1085, 1091, and 1097; and (b) a light chain variable (V.sub.L) domain comprising a LCDR1 comprising or consisting essentially of the amino acid sequences as shown in any one of SEQ ID NOs: 847, 853, 860, 866, 871, 888, 894, 900, 906, 912, 918, 924, 930, 936, 942, 947, 953, 959, 965, 971, 977, 983, 989, 995, 1001, 1007, 1013, 1019, 1025, 1032, 1038, 1047, 1053, 1059, 1068, 1074, 1080, 1086, 1092, and 1098, a LCDR2 comprising or consisting essentially of the amino acid sequences as shown in any one of SEQ ID NOs: 848, 854, 861, 867, 883, 889, 895, 901, 907, 913, 919, 925, 931, 937, 948, 954, 960, 966, 972, 978, 984, 990, 996, 1002, 1008, 1014, 1020, 1026, 1033, 1039, 1048, 1054, 1060, 1069, 1075, 1081, 1087, 1093, and 1099, and a LCDR3 comprising or consisting essentially of the amino acid sequences as shown in any one of SEQ ID NOs: 849, 855, 862, 868, 884, 890, 896, 902, 908, 914, 920, 926, 932, 938, 949, 955, 961, 967, 973, 979, 985, 991, 997, 1003, 1009, 1015, 1021, 1027, 1034, 1040, 1049, 1055, 1061, 1070, 1076, 1082, 1088, 1094, and 1100.
[0378] The Shiga toxin effector polypeptides, the Shiga toxin effector polypeptide scaffolds, and cell-targeting molecules of the present invention may optionally be conjugated to one or more additional agents, which may include therapeutic agents, diagnostic agents, and/or other additional exogenous materials known in the art, including such agents as described herein. In certain embodiments, the Shiga toxin effector polypeptide, the Shiga toxin effector polypeptide scaffold or cell-targeting molecule of the present invention is PEGylated or albuminated, such as, e.g., to provide de-immunization, disrupt furin-cleavage by masking the extended loop and/or the furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment derived region, improve pharmacokinetic properties, and/or improve immunogenicity (see e.g., Wang Q et al., Cancer Res 53: 4588-94 (1993); Tsutsumi Y et al., Proc Natl Acad Sci USA 97: 8548-53 (2000); Buse J, El-Aneed A, Nanomed 5: 1237-60 (2010); Lim S et. al., J Control Release 207-93 (2015)).
III. General Functions of the Cell-Targeting Molecules of the Present Invention
[0379] The molecules of the present invention are useful in a variety of applications involving cell-targeting molecules (see e.g. WO 2014/164680, WO 2014/164693, WO 2015/113005, WO 2015/113007, WO 2015/120058, WO 2015/138435, WO 2015/138452, US 2015/0259428, WO 2015/191764, US2016/177284, WO 2016/126950, WO 2016/196344, and WO 2017/019623). The Shiga toxin effector polypeptides, binding region polypeptides, and linkers of the present invention may be used as components to create cell-targeting molecules with improved properties (see e.g. WO 2014/164680, WO 2014/164693, WO 2015/113005, WO 2015/113007, WO 2015/120058, WO 2015/138435, WO 2015/138452, US 2015/0259428, WO 2015/191764, US2016/177284, WO 2016/126950, WO 2016/196344, and WO 2017/019623). The Shiga toxin effector polypeptides, binding region polypeptides, and linkers of the present invention are useful as components of various therapeutic and/or diagnostic molecules, such as, e.g. ligand-toxin fusions, immunotoxins, and/or immuno-conjugates (see e.g. WO 2015/113005, WO 2015/113007, WO 2015/138435, US 2015/0259428, WO 2015/191764, US2016/177284, WO 2016/126950, WO 2016/196344, and WO 2017/019623). The functional association of Shiga toxin effector polypeptides with cell-targeting binding regions enables the creation of cell-targeting molecules which selectively kill, inhibit the growth of, deliver exogenous material to, and/or detect specific cell types. For example, certain cell-targeting molecules of the present invention may be potently cytotoxic to target-expressing cells via their abilities to efficiently deliver into the interior of a target-expressing cell a catalytically active, Shiga toxin effector polypeptide(s) that is able to effectively route to the cytosol.
[0380] The Shiga toxin effector polypeptides and cell-targeting molecules of the present invention are useful in diverse applications involving, e.g., cell-killing; cell growth inhibition; intracellular, cargo delivery; biological information gathering; immune response stimulation, and/or remediation of a health condition. The cell-targeting molecules of the present invention are useful as therapeutic and/or diagnostic molecules, such as, e.g., as cell-targeting, cytotoxic, therapeutic molecules; cell-targeting, nontoxic, delivery vehicles; and/or cell-targeting, diagnostic molecules; for examples in applications involving the in vivo targeting of specific cell types for the diagnosis or treatment of a variety of diseases, including cancers, immune disorders, and microbial infections.
[0381] The Shiga toxin effector polypeptides of the present invention allow for controlled and site-specific conjugation of molecular cargos to form cell-targeting molecules capable of cell-targeted delivery of their cargo(s). For example, conjugation can be chemically limited to occur at a single amino acid residue or linker attached thereto in order to produce a homogeneous product with a defined conjugate stoichiometry using strategies such as engineered cysteine residues, unnatural amino acid residues, and/or enzymatic conjugation. Similarly, the cell-targeting molecules of the present invention allow for controlled and site-specific conjugation of heterologous molecules to form useful cell-targeting molecules conjugates, such as, e.g., conjugates involving therapeutic molecules, immunogenicity-reducing agents, half-life extending agents, and various other cargos.
[0382] The cell-targeting molecules of the present invention, and compositions thereof, have uses, e.g., for the selective delivery of cargos to target-expressing cells and as therapeutics for the treatment of a variety of diseases, disorders, and conditions, which include genetic disorders, genetic predispositions, infections, cancers, tumors, growth abnormalities, and/or immune disorders. Certain cell-targeting molecules of the present invention, and compositions thereof, may be used to selectively deliver conjugated cargo(s) to a target-expressing cell type(s) in the presence of one or more other cell types based on its cell-targeting and cellular internalization activity(ies), such as, e.g., a cargo having a desired, intracellular function. In addition, certain cell-targeting molecules of the present invention, and compositions thereof, may be used to selectively kill a target-expressing cell in the presence of one or more other cell types based on its cell-targeting activity and cellular internalization activity(ies), such as, e.g., by delivering into the interior of the targeted, target-expressing cell a component of the cell-targeting molecule which is cytotoxic at an intracellular location.
[0383] Depending on the embodiment, a Shiga toxin effector polypeptide or cell-targeting molecule of the present invention may have or provide one or more of the following characteristics or functionalities: (1) de-immunization (see e.g. WO 2015/113005; WO 2015/113007), (2) protease-cleavage resistance (see e.g. WO 2015/191764), (3) potent cytotoxicity at certain concentrations, (4) intracellular delivery of a cargo consisting of an additional material (e.g. a heterologous, T-cell epitope) (see e.g. WO 2015/113005), (4) selective cytotoxicity, (6) low off-target toxicity in multicellular organisms at certain doses or dosages (see e.g. WO 2015/191764), (7) delivery of a heterologous, T-cell epitope to the MHC class I presentation pathway of a target cell (see e.g. WO 2015/113005), and/or (8) stimulation of CD8+ T-cell immune response(s). Certain embodiments of the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention are multi-functional because the molecules have two or more of the characteristics or functionalities described herein.
[0384] In certain embodiments, the cell-targeting molecules of the present invention are capable of binding extracellular target biomolecules associated with the cell surface of particular cell types and entering those cells. Once internalized within a targeted cell type, certain embodiments of the cell-targeting molecules of the invention are capable of routing an enzymatically active, cytotoxic, Shiga toxin effector polypeptide fragment into the cytosol of the target cell and eventually killing the cell. Alternatively, nontoxic or reduced-toxicity variants of the cell-targeting molecules of the present invention may be used to deliver additional exogenous materials into target cells, such as B-cell or T-cell epitopes, peptides, proteins, polynucleotides, and detection-promoting agents. This system is modular, in that any number of diverse binding regions can be used to target a Shiga toxin effector polypeptide of the present invention to various, diverse cell types.
A. Cell-Kill Via Shiga Toxin A Subunit Cytotoxicity
[0385] Certain embodiments of the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention are cytotoxic. Certain further embodiments of the cell-targeting molecules of the present invention are cytotoxic only due to the presence of one or more Shiga toxin effector polypeptide components. The A Subunits of members of the Shiga toxin family each comprise an enzymatically active polypeptide region capable of killing a eukaryotic cell once in the cell's cytosol. Because members of the Shiga toxin family are adapted to killing eukaryotic cells, molecules derived from Shiga toxins, such as, e.g., molecules comprising certain embodiments of the Shiga toxin effector polypeptides of the present invention can exhibit potent cell-kill activities.
[0386] For certain embodiments of the cell-targeting molecules of the present invention, upon contacting a cell physically coupled with an extracellular target biomolecule of the binding region of the cell-targeting molecule (e.g. a target positive cell), the cell-targeting molecule is capable of causing death of the cell. For certain further embodiments, the CD.sub.50 value of the cell-targeting molecule is less than 5, 2.5, 1, 0.5, or 0.25 nM, which is vastly more potent than an untargeted, wild-type, Shiga toxin effector polypeptide (e.g. SEQ ID NO:830).
[0387] Cell-kill may be accomplished using a molecule of the present invention under varied conditions of target cells, such as, e.g., an ex vivo manipulated target cell, a target cell cultured in vitro, a target cell within a tissue sample cultured in vitro, or a target cell in an in vivo setting like within a multicellular organism.
[0388] In certain embodiments, the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention comprise (1) a de-immunized, Shiga toxin effector sub-region, (2) a protease-cleavage resistant region near the carboxy-terminus of a Shiga toxin A1 fragment derived region, (3) a carboxy-terminal, endoplasmic reticulum retention/retrieval signal motif; and/or (4) a heterologous, T-cell epitope embedded or inserted region; however, for certain further embodiments, these structural modifications do not significantly alter the potency of Shiga toxin cytotoxicity as compared to a reference molecules comprising a wild-type Shiga toxin A Subunit polypeptide, such as, e.g., a wild-type Shiga toxin A1 fragment. Thus, Shiga toxin effector polypeptides and cell-targeting molecules of the present invention which are de-immunized, protease cleavage resistant, and/or carrying embedded or inserted, heterologous, epitopes can maintain potent cytotoxicity while providing one or more various other functionalities or properties.
[0389] Already cytotoxic cell-targeting molecules comprising Shiga toxin effector polypeptides may be engineered by the skilled worker using the information and methods provided herein to be more cytotoxic and/or to have redundant, backup cytotoxicities operating via completely different mechanisms. These multiple cytotoxic mechanisms may complement each other by their diversity of functions (such as by providing potent killing via two mechanisms of cell-killing, direct and indirect, as well as mechanisms of immuno-stimulation to the local area), redundantly backup each other (such as by providing one cell-killing mechanism in the absence of the other mechanismslike if a target cell is resistant to or acquires some immunity to a subset of previously active mechanisms), and/or protect against developed resistance (by limiting resistance to the less probable situation of the malignant or infected cell blocking multiple, different cell-killing mechanisms simultaneously).
B. Delivery of a T-Cell Epitope for MHC Class I Presentation on a Cell Surface
[0390] In certain embodiments, the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention comprise a T-cell epitope, which enables the engineering of T-cell epitope delivering molecules with virtually unlimited choices of epitope-peptide cargos for delivery and cell-surface presentation by a nucleated, chordate cell. For certain embodiments, the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention are each capable of delivering one or more T-cell epitopes, associated with the Shiga toxin effector polypeptides and/or cell-targeting molecules, to the proteasome of a cell. The delivered T-cell epitope are then proteolytic processed and presented by the MHC class I pathway on the surface of the cell. By conjugating MHC class I epitopes to cell-targeting molecules, the targeted delivery and presentation of immuno-stimulatory antigens may be accomplished in order to harness and direct a beneficial function(s) of a chordate immune system.
[0391] For certain embodiments, the Shiga toxin effector polypeptide or cell-targeting molecule of the present invention is capable of delivering a T-cell epitope to a MHC class I molecule of a cell for cell-surface presentation. In certain embodiments, the Shiga toxin effector polypeptide or cell-targeting molecule of the present invention comprises a heterologous, T-cell epitope, whether as an additional exogenous material or embedded or inserted within a Shiga toxin effector polypeptide. For certain further embodiments, the Shiga toxin effector polypeptide or cell-targeting molecule of the present invention is capable of delivering an embedded or inserted T-cell epitope to a MHC class I molecule for cell-surface presentation.
[0392] For certain embodiments, the Shiga toxin effector polypeptide of the present invention is capable of delivering a T-cell epitope, which is conjugated to the Shiga toxin effector polypeptide, to a MHC class I molecule of a cell in which the Shiga toxin effector polypeptide is present for presentation of the T-cell epitope by the MHC class I molecule on a surface of the cell. For certain further embodiments, the T-cell epitope is a heterologous, T-cell epitope. For certain further embodiments, the T-cell epitope functions as CD8+ T-cell epitope, whether already known or identified in the future using methods which are currently routine to the skilled worker.
[0393] For certain embodiments, the cell-targeting molecule of the present invention is capable of delivering a T-cell epitope, which is associated with the cell-targeting molecule, to a MHC class I molecule of a cell for presentation of the T-cell epitope by the MHC class I molecule on a surface of the cell. For certain further embodiments, the T-cell epitope is a heterologous, T-cell epitope which is conjugated to the Shiga toxin effector polypeptide. For certain further embodiments, the T-cell epitope functions as CD8+ T-cell epitope, whether already known or identified in the future using methods which are currently routine to the skilled worker.
[0394] For certain embodiments, upon contacting a cell with the cell-targeting molecule of the present invention, the cell-targeting molecule is capable of delivering a T-cell epitope-peptide, which is associated with the cell-targeting molecule, to a MHC class I molecule of the cell for presentation of the T-cell epitope-peptide by the MHC class I molecule on a surface of the cell. For certain further embodiments, the T-cell epitope-peptide is a heterologous epitope which is conjugated to a Shiga toxin effector polypeptide. For certain further embodiments, the T-cell epitope-peptide functions as CD8+ T-cell epitope, whether already known or identified in the future using methods which are currently routine to the skilled worker.
[0395] The addition of a heterologous epitope into or presence of a heterologous epitope in a cell-targeting molecule of the present invention, whether as an additional exogenous material or embedded or inserted within a Shiga toxin effector polypeptide, enables methods of using such cell-targeting molecules for the cell-targeted delivery of a chosen epitope for cell-surface presentation by a nucleated, target cell within a chordate.
[0396] One function of certain, CD8+ T-cell hyper-immunized, Shiga toxin effector polypeptides and cell-targeting molecules of the present invention is the delivery of one or more T-cell epitope-peptides to a MHC class I molecule for MHC class I presentation by a cell. Delivery of exogenous, T-cell epitope-peptides to the MHC class I system of a target cell can be used to induce the target cell to present the T-cell epitope-peptide in association with MHC class I molecules on the cell surface, which subsequently leads to the activation of CD8+ effector T-cells to attack the target cell.
[0397] The skilled worker, using techniques known in the art, can associate, couple, and/or link certain, Shiga toxin effector polypeptides of the present invention to various other cell-targeting binding regions to create cell-targeting molecules of the present invention which target specific, extracellular, target biomolecules physically coupled to cells and promote target-cell internalization of these cell-targeting molecules. All nucleated vertebrate cells are believed to be capable of presenting intracellular epitopes using the MHC class I system. Thus, extracellular target biomolecules of the cell-targeting molecules of the invention may in principle target any nucleated vertebrate cell for T-cell epitope delivery to a MHC class I presentation pathway of such a cell.
[0398] The epitope-delivering functions of the Shiga toxin effector polypeptides and cell-targeting molecules of the present invention can be detected and monitored by a variety of standard methods known in the art to the skilled worker and/or described herein (see e.g. WO 2015/113005).
[0399] Certain assays to monitor this function of the polypeptides and molecules of the present invention involve the direct detection of a specific MHC class I/peptide antigen complex in vitro or ex vivo. Common methods for direct visualization and quantitation of peptide-MHC class I complexes involve various immuno-detection reagents known to the skilled worker. For example, specific monoclonal antibodies can be developed to recognize a particular MHC/class I/peptide antigen complex. Similarly, soluble, multimeric T cell receptors, such as the TCR-STAR reagents (Altor Bioscience Corp., Mirmar, FL, U.S.) can be used to directly visualize or quantitate specific MHC I/antigen complexes (Zhu X et al., J Immunol 176: 3223-32 (2006)). These specific mAbs or soluble, multimeric T-cell receptors may be used with various detection methods, including, e.g. immunohistochemistry, flow cytometry, and enzyme-linked immuno assay (ELISA).
[0400] An alternative method for direct identification and quantification of MHC I/peptide complexes involves mass spectrometry analyses, such as, e.g., the ProPresent Antigen Presentation Assay (ProImmune, Inc., Sarasota, FL, U.S.) in which peptide-MCH class I complexes are extracted from the surfaces of cells, then the peptides are purified and identified by sequencing mass spectrometry (Falk K et al., Nature 351: 290-6 (1991)).
[0401] In certain assays to monitor the T-cell epitope delivery and MHC class I presentation function of the polypeptides and molecules of the present invention involve computational and/or experimental methods to monitor MHC class I and peptide binding and stability. Several software programs are available for use by the skilled worker for predicting the binding responses of peptides to MHC class I alleles, such as, e.g., The Immune Epitope Database and Analysis Resource (IEDB) Analysis Resource MHC-I binding prediction Consensus tool (Kim Y et al., Nucleic Acid Res 40: W525-30 (2012). Several experimental assays have been routinely applied, such as, e.g., cell surface binding assays and/or surface plasmon resonance assays to quantify and/or compare binding kinetics (Miles K et al., Mol Immunol 48: 728-32 (2011)). Additionally, other MHC-peptide binding assays based on a measure of the ability of a peptide to stabilize the ternary MHC-peptide complex for a given MHC class I allele, as a comparison to known controls, have been developed (e.g., MHC-peptide binding assay from ProImmune, Inc.).
[0402] Alternatively, measurements of the consequence of MHC class I/peptide antigen complex presentation on the cell surface can be performed by monitoring the cytotoxic T-cell (CTL) response to the specific complex. These measurements by include direct labeling of the CTLs with MHC class I tetramer or pentamer reagents. Tetramers or pentamers bind directly to T cell receptors of a particular specificity, determined by the Major Histocompatibility Complex (MHC) allele and peptide complex. Additionally, the quantification of released cytokines, such as interferon gamma or interleukins by ELISA or enzyme-linked immunospot (ELIspot) is commonly assayed to identify specific CTL responses. The cytotoxic capacity of CTL can be measured using a number of assays, including the classical 51 Chromium (Cr) release assay or alternative non-radioactive cytotoxicity assays (e.g., CytoTox96 non-radioactive kits and CellTox CellTiter-GLO kits available from Promega Corp., Madison, WI, U.S.), Granzyme B ELISpot, Caspase Activity Assays or LAMP-1 translocation flow cytometric assays. To specifically monitor the killing of target cells, carboxyfluorescein diacetate succinimidyl ester (CF SE) can be used to easily and quickly label a cell population of interest for in vitro or in vivo investigation to monitor killing of epitope specific CSFE labeled target cells (Durward M et al., J Vis Exp 45 pii 2250 (2010)).
[0403] In vivo responses to WIC class I presentation can be followed by administering a MHC class I/antigen promoting agent (e.g., an immunogenic peptide, protein or inactivated/attenuated virus vaccine) followed by challenge with an active agent (e.g. a virus) and monitoring responses to that agent, typically in comparison with unvaccinated controls. Ex vivo samples can be monitored for CTL activity with methods similar to those described previously (e.g. CTL cytotoxicity assays and quantification of cytokine release).
[0404] HLA-A, HLA-B, and/or HLA-C molecules are isolated from the intoxicated cells after lysis using immune affinity (e.g., an anti-WIC antibody pulldown purification) and the associated peptides (i.e., the peptides presented by the isolated WIC molecules) are recovered from the purified complexes. The recovered peptides are analyzed by sequencing mass spectrometry. The mass spectrometry data is compared against a protein database library consisting of the sequence of the exogenous (non-self) peptide (T-cell epitope X) and the international protein index for humans (representing self or non-immunogenic peptides). The peptides are ranked by significance according to a probability database. All detected antigenic (non-self) peptide sequences are listed. The data is verified by searching against a scrambled decoy database to reduce false hits (see e.g. Ma B, Johnson R, Mol Cell Proteomics 11: O111.014902 (2012)). The results will demonstrate that peptides from the T-cell epitope X are presented in WIC complexes on the surface of intoxicated target cells.
[0405] The set of presented peptide-antigen-WIC complexes can vary between cells due to the antigen-specific HLA molecules expressed. T-cells can then recognize specific peptide-antigen-MHC complexes displayed on a cell surface using different TCR molecules with different antigen-specificities.
[0406] Because multiple T-cell epitopes may be delivered by a cell-targeting molecule of the invention, such as, e.g., by embedding two or more different T-cell epitopes in a single proteasome delivering effector polypeptide, a single cell-targeting molecule of the invention may be effective chordates of the same species with different MHC class variants, such as, e.g., in humans with different HLA alleles. This may allow for the combining within a single molecule of different T-cell epitopes with different effectiveness in different sub-populations of subjects based on WIC complex protein diversity and polymorphisms. For example, human MHC complex proteins, HLA proteins, vary among humans based on genetic ancestry, e.g. African (sub-Saharan), Amerindian, Caucasiod, Mongoloid, New Guinean and Australian, or Pacific islander.
[0407] The applications involving the T-cell epitope delivering polypeptides and molecules of the present invention are vast. Every nucleated cell in a mammalian organism may be capable of MHC class I pathway presentation of immunogenic, T-cell epitope-peptides on their cell outer surfaces complexed to MHC class I molecules. In addition, the sensitivity of T-cell epitope recognition is so exquisite that only a few MHC-I peptide complexes are required to be presented to result in an immune response, e.g., even presentation of a single complex can be sufficient for recognition by an effector T-cell (Sykulev Y et al., Immunity 4: 565-71 (1996)).
[0408] The activation of T-cell responses are desired characteristics of certain anti-cancer, anti-neoplastic, anti-tumor, and/or anti-microbial biologic drugs to stimulate the patient's own immune system toward targeted cells. Activation of a robust and strong T-cell response is also a desired characteristic of many vaccines. The presentation of a T-cell epitope by a target cell within an organism can lead to the activation of robust immune responses to a target cell and/or its general locale within an organism. Thus, the targeted delivery of a T-cell epitope for presentation may be utilized for as a mechanism for activating T-cell responses during a therapeutic regime.
[0409] The presentation of a T-cell immunogenic epitope-peptide by the WIC class I system targets the presenting cell for killing by CTL-mediated lysis and also triggers immune stimulation in the local microenvironment. By engineering immunogenic epitope sequences within Shiga toxin effector polypeptide components of target-cell-internalizing therapeutic molecules, the targeted delivery and presentation of immuno-stimulatory antigens may be accomplished. The presentation of immuno-stimulatory non-self antigens, such as e.g. known viral antigens with high immunogenicity, by target cells signals to other immune cells to destroy the target cells as well as to recruit more immune cells to the area.
[0410] The presentation of an immunogenic, T-cell epitope-peptide by the MHC class I complex targets the presenting cell for killing by CTL-mediated cytolysis. The presentation by targeted cells of immuno-stimulatory non-self antigens, such as, e.g., known viral epitope-peptides with high immunogenicity, can signal to other immune cells to destroy the target cells and recruit more immune cells to the target cell site within a chordate.
[0411] Thus, already cytotoxic molecules, such as e.g. therapeutic or potentially therapeutic molecules comprising Shiga toxin effector polypeptides, may be engineered using methods of the present invention into more cytotoxic molecules and/or to have an additional cytotoxic mechanism operating via delivery of a T-cell epitope, presentation, and stimulation of effector T-cells. These multiple cytotoxic mechanisms may complement each other (such as by providing both direct target-cell-killing and indirect (CTL-mediated) cell-killing, redundantly backup each other (such as by providing one mechanism of cell-killing in the absence of the other), and/or protect against the development of therapeutic resistance (by limiting resistance to the less probable situation of the malignant or infected cell evolving to block two different cell-killing mechanisms simultaneously).
[0412] In addition, a cytotoxic molecule comprising a Shiga toxin effector polypeptide component that exhibits catalytic-based cytotoxicity may be engineered by the skilled worker using routine methods into enzymatically inactive variants. For example, the cytotoxic Shiga toxin effector polypeptide component of a cytotoxic molecule may be conferred with reduced activity and/or rendered inactive by the introduction of one or mutations and/or truncations such that the resulting molecule can still be cytotoxic via its ability to deliver a T-cell epitope to the MHC class I system of a target cell and subsequent presentation to the surface of the target cell. In another example, a T-cell epitope may be inserted or embedded into a Shiga toxin effector polypeptide such that the Shiga toxin effector polypeptide is inactivated by the added epitope (see e.g. WO 2015/113005). This approach removes one cytotoxic mechanism while retaining or adding another and may also provide a molecule capable of exhibiting immuno-stimulation to the local area of a target cell(s) within an organism via delivered T-cell epitope presentation or antigen seeding. Furthermore, non-cytotoxic variants of the cell-targeting molecules of the present invention which comprise embedded or inserted, heterologous, T-cell epitopes may be useful in applications involving immune-stimulation within a chordate and/or labeling of target cells within a chordate with MHC class I molecule displayed epitopes.
[0413] The ability to deliver a T-cell epitope of certain Shiga toxin effector polypeptides and cell-targeting molecules of the present invention may be accomplished under varied conditions and in the presence of non-targeted bystander cells, such as, e.g., an ex vivo manipulated target cell, a target cell cultured in vitro, a target cell within a tissue sample cultured in vitro, or a target cell in an in vivo setting like within a multicellular organism.
C. Cell-Kill Via Targeted Cytotoxicity and/or Engagement of Cytotoxic T-Cells
[0414] For certain embodiments, the cell-targeting molecule of the present invention can provide 1) delivery of a T-cell epitope for MHC class I presentation by a target cell and/or 2) potent cytotoxicity. For certain embodiments of the cell-targeting molecules of the present invention, upon contacting a cell physically coupled with an extracellular target biomolecule of the cell-targeting binding region, the cell-targeting molecule of the invention is capable of causing death of the cell. The mechanism of cell-kill may be direct, e.g. via the enzymatic activity of a toxin effector polypeptide region, or indirect via CTL-mediated cytolysis.
1. Indirect Cell-Kill Via T-Cell Epitope Delivery and MHC Class I Presentation
[0415] Certain embodiments of the cell-targeting molecules of the present invention are cytotoxic because they comprise a CD8+ T-cell epitope capable of being delivered to the MHC class I presentation pathway of a target cell and presented on a cellular surface of the target cell. For example, T-cell epitope delivering, Shiga toxin effector polypeptides of the present invention, with or without endogenous epitope de-immunization, may be used as components of cell-targeting molecules for applications involving indirect cell-killing (see e.g. WO 2015/113005).
[0416] In certain embodiments of the cell-targeting molecules of the present invention, upon contacting a cell physically coupled with an extracellular target biomolecule of the cell-targeting binding region, the cell-targeting molecule of the invention is capable of indirectly causing the death of the cell, such as, e.g., via the presentation of one or more T-cell epitopes by the target cell and the subsequent recruitment of CTLs which kill the target cell.
[0417] The presentation of an antigenic peptide complexed with a MHC class I molecule by a cell sensitizes the presenting cell to targeted killing by cytotoxic T-cells (CTLs) via the induction of apoptosis, lysis, and/or necrosis. In addition, the CTLs which recognize the target cell may release immuno-stimulatory cytokines, such as, e.g., interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF), macrophage inflammatory protein-1 beta (MIP-1beta), and interleukins such as IL-17, IL-4, and IL-22. Furthermore, CTLs activated by recognition of a presented epitope may indiscriminately kill other cells proximal to the presenting cell regardless of the peptide-MHC class I complex repertoire presented by those proximal cells (Wiedemann A et al., Proc Natl Acad Sci USA 103: 10985-90 (2006)).
[0418] Because of MHC allele diversity within different species, a cell-targeting molecule of the present invention comprising only a single epitope may exhibit varied effectiveness to different patients or subjects of the same species. However, certain embodiments of the cell-targeting molecules of the present invention may each comprise multiple, T-cell epitopes that are capable of being delivered to the MHC class I system of a target cell simultaneously. Thus, for certain embodiments of the cell-targeting molecules of the present invention, a cell-targeting molecule is used to treat different subjects with considerable differences in their MHC molecules' epitope-peptide binding affinities (i.e. considerable differences in their MHC alleles and/or MHC genotypes). In addition, certain embodiments of the cell-targeting molecules of the present invention reduce or prevent target cell adaptations to escape killing (e.g. a target cancer cell mutating to escape therapeutic effectiveness or mutant escape) by using multiple cell-killing mechanisms simultaneously (e.g. direct killing and indirect killing via multiple different T-cell epitopes simultaneously).
2. Direct Cell-Kill Via Cell-Targeted, Shiga Toxin Cytotoxicity
[0419] Certain embodiments of the cell-targeting molecules of the present invention are cytotoxic because they comprise a catalytically active, Shiga toxin effector polypeptide and regardless of the presence of any cytotoxic agent or immunogenic, CD8+ T-cell epitope in the molecule. For example, Shiga toxin effector polypeptides of the present invention, with or without endogenous epitope de-immunization, may be used as components of cell-targeting molecules for applications involving direct cell-killing, such as, e.g., via the ribotoxic, enzymatic activity of a Shiga toxin effector polypeptide or ribosome binding and interference with ribosome function due to a non-catalytic mechanism(s). For certain embodiments of the CD8+ T-cell hyper-immunized, cell-targeting molecules of the present invention, upon contacting a cell physically coupled with an extracellular target biomolecule of the cell-targeting binding region, the cell-targeting molecule of the invention is capable of directly causing the death of the cell, such as, e.g., without the involvement of an untargeted, cytotoxic T-cell (see Section III-D, supra).
D. Selective Cytotoxicity Among Cell Types
[0420] Certain cell-targeting molecules of the present invention have uses in the selective killing of specific target cells in the presence of untargeted, bystander cells. By targeting the delivery of Shiga toxin effector polypeptides of the present invention to specific cells via a cell-targeting binding region(s), the cell-targeting molecules of the present invention can exhibit cell-type specific, restricted cell-kill activities resulting in the exclusive or preferential killing selected cell types in the presence of untargeted cells. Similarly, by targeting the delivery of immunogenic T-cell epitopes to the MHC class I pathway of target cells, the subsequent presentation of T-cell epitopes and CTL-mediated cytolysis of target cells induced by the cell-targeting molecules of the invention can be restricted to exclusively or preferentially killing selected cell types in the presence of untargeted cells. In addition, both the cell-targeted delivery of a cytotoxic, Shiga toxin effector polypeptide region and an immunogenic, T-cell epitope can be accomplished by a single cell-targeting molecule of the present invention such that deliver of both potentially cytotoxic components is restricted exclusively or preferentially to target cells in the presence of untargeted cells.
[0421] For certain embodiments, the cell-targeting molecule of the present invention is cytotoxic at certain concentrations. In certain embodiments, upon administration of the cell-targeting molecule of the present invention to a mixture of cell types, the cytotoxic cell-targeting molecule is capable of selectively killing those cells which are physically coupled with an extracellular target biomolecule compared to cell types not physically coupled with an extracellular target biomolecule. For certain embodiments, the cytotoxic cell-targeting molecule of the present invention is capable of selectively or preferentially causing the death of a specific cell type within a mixture of two or more different cell types. This enables targeting cytotoxic activity to specific cell types with a high preferentiality, such as a 3-fold cytotoxic effect, over bystander cell types that do not express the target biomolecule. Alternatively, the expression of the target biomolecule of the binding region may be non-exclusive to one cell type if the target biomolecule is expressed in low enough amounts and/or physically coupled in low amounts with cell types that are not to be targeted. This enables the targeted cell-killing of specific cell types with a high preferentiality, such as a 3-fold cytotoxic effect, over bystander cell types that do not express significant amounts of the target biomolecule or are not physically coupled to significant amounts of the target biomolecule.
[0422] For certain further embodiments, upon administration of the cytotoxic cell-targeting molecule to two different populations of cell types, the cytotoxic cell-targeting molecule is capable of causing cell death as defined by the half-maximal cytotoxic concentration (CD50) on a population of target cells, whose members express an extracellular target biomolecule of the binding region of the cytotoxic cell-targeting molecule, at a dose at least three-times lower than the CD.sub.50 dose of the same cytotoxic cell-targeting molecule to a population of cells whose members do not express an extracellular target biomolecule of the binding region of the cytotoxic cell-targeting molecule.
[0423] For certain embodiments, the cytotoxic activity of a cell-targeting molecule of the present invention toward populations of cell types physically coupled with an extracellular target biomolecule is at least 3-fold higher than the cytotoxic activity toward populations of cell types not physically coupled with any extracellular target biomolecule of the binding region. According to the present invention, selective cytotoxicity may be quantified in terms of the ratio (a/b) of (a) cytotoxicity towards a population of cells of a specific cell type physically coupled with a target biomolecule of the binding region to (b) cytotoxicity towards a population of cells of a cell type not physically coupled with a target biomolecule of the binding region. In certain embodiments, the cytotoxicity ratio is indicative of selective cytotoxicity which is at least 3-fold, 5-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 40-fold, 50-fold, 75-fold, 100-fold, 250-fold, 500-fold, 750-fold, or 1000-fold higher for populations of cells or cell types physically coupled with a target biomolecule of the binding region compared to populations of cells or cell types not physically coupled with a target biomolecule of the binding region.
[0424] For certain embodiments, the preferential cell-killing function or selective cytotoxicity of a cell-targeting molecule of the present invention is due to an additional exogenous material (e.g. a cytotoxic material) and/or heterologous, T-cell epitope present in a Shiga toxin effector polypeptide of the present invention and not necessarily a result of the catalytic activity of a Shiga toxin effector polypeptide.
[0425] This preferential cell-killing function allows a targeted cell to be killed by certain cytotoxic, cell-targeting molecules of the present invention under varied conditions and in the presence of non-targeted bystander cells, such as ex vivo manipulated mixtures of cell types, in vitro cultured tissues with mixtures of cell types, or in vivo in the presence of multiple cell types (e.g. in situ or in a native location within a multicellular organism).
E. Delivery of Molecular Cargos into Interior Compartments of Target Cells
[0426] In addition to cytotoxic, cytostatic, and immune stimulation applications, cell-targeting molecules of the present invention optionally may be used for targeted intracellular delivery functions, such as, e.g., in applications involving information gathering and diagnostic functions.
[0427] Because the cell-targeting molecules of the invention, including reduced cytotoxicity and/or nontoxic forms thereof, are capable of entering cells physically coupled with an extracellular target biomolecule recognized by the cell-targeting molecule's binding region, certain embodiments of the cell-targeting molecules of the invention may be used to deliver additional exogenous materials or cargos (e.g. conjugated molecules) into the interior of targeted cell types. For example, non-toxic variants of the cytotoxic, cell-targeting molecules of the invention, or optionally cytotoxic variants, may be used to deliver additional exogenous materials to and/or label the interiors of cells physically coupled with an extracellular target biomolecule of the binding region of the cell-targeting molecule. Various types of cells and/or cell populations which express target biomolecules to at least one cellular surface may be targeted by the cell-targeting molecules of the invention for receiving exogenous materials. The functional components of the present invention are modular, in that various Shiga toxin effector polypeptides, additional exogenous materials, and binding regions may be associated with each other to provide cell-targeting molecules suitable for diverse applications involving cargo delivery, such as, e.g., non-invasive, in vivo imaging of tumor cells.
[0428] This delivery of exogenous material function of certain cell-targeting molecules of the present invention may be accomplished under varied conditions and in the presence of non-targeted bystander cells, such as, e.g., an ex vivo manipulated target cell, a target cell cultured in vitro, a target cell within a tissue sample cultured in vitro, or a target cell in an in vivo setting like within a multicellular organism. Furthermore, the selective delivery of exogenous material to certain cells by certain cell-targeting molecules of the present invention may be accomplished under varied conditions and in the presence of non-targeted bystander cells, such as ex vivo manipulated mixtures of cell types, in vitro cultured tissues with mixtures of cell types, or in vivo in the presence of multiple cell types (e.g. in situ or in a native location within a multicellular organism).
[0429] Shiga toxin effector polypeptides and cell-targeting molecules which are not capable, such as a certain concentration ranges, of killing a target cell and/or delivering an embedded or inserted epitope for cell-surface presentation by a WIC molecule of a target cell may still be useful for delivering exogenous materials into cells, such as, e.g., detection-promoting agents.
[0430] For certain embodiments, the Shiga toxin effector polypeptides of the present invention exhibits low to zero cytotoxicity and thus are referred to herein as noncytotoxic and/or reduced cytotoxic. For certain embodiments, the cell-targeting molecule of the present invention exhibits low to zero cytotoxicity and may be referred to as noncytotoxic and/or reduced cytotoxic variants. For example, certain embodiments of the molecules of the present invention do not exhibit a significant level of Shiga toxin based cytotoxicity wherein at doses of less than 1,000 nM, 500 nM, 100 nM, 75 nM, 50 nM, there is no significant amount of cell death as compared to the appropriate reference molecule, such as, e.g., as measured by an assay known to the skilled worker and/or described herein. For certain further embodiments, the molecules of the present invention do not exhibit any toxicity at dosages of 1-100 microgram (m) per kilogram (kg) of a mammalian recipient. Reduced-cytotoxic variants may still be cytotoxic at certain concentrations or dosages but exhibit reduced cytotoxicity, such as, e.g., are not capable of exhibiting a significant level of Shiga toxin cytotoxicity in certain situations.
[0431] Shiga toxin effector polypeptides of the present invention, and certain cell-targeting molecules comprising the same, can be rendered non-cytotoxic, such as, e.g., via the addition of one or more amino acid substitutions known to the skilled worker to inactivate a Shiga toxin A Subunit and/or Shiga toxin effector polypeptide, including exemplary substitutions described herein. The non-cytotoxic and reduced cytotoxic variants of the cell-targeting molecules of the present invention may be in certain situations more suitable for delivery of additional exogenous materials than more cytotoxic variants.
F. Information Gathering for Diagnostic Functions
[0432] Certain cell-targeting molecules of the present invention have uses in the in vitro and/or in vivo detection of specific cells, cell types, and/or cell populations, as well as specific subcellular compartments of any of the aforementioned. Reduced-cytotoxicity and/or nontoxic forms of the cytotoxic, cell-targeting molecules of the invention that are conjugated to detection-promoting agents optionally may be used for diagnostic functions, such as for companion diagnostics used in conjunction with a therapeutic regimen comprising the same or a related binding region, such as, e.g., a binding region with high-affinity binding to the same target biomolecule, an overlapping epitope, and/or the same epitope.
[0433] In certain embodiments, the cell-targeting molecules described herein are used for both diagnosis and treatment, or for diagnosis alone. When the same cytotoxic cell-targeting molecule is used for both diagnosis and treatment, for certain embodiments of the present invention the cell-targeting molecule variant which incorporates a detection-promoting agent for diagnosis may have its cytotoxicity reduced or may be rendered nontoxic by catalytic inactivation of its Shiga toxin effector polypeptide region(s) via one or more amino acid substitutions, including exemplary substitutions described herein. For example, certain nontoxic variants of the cell-targeting molecules of the present invention exhibit less than 5%, 4%, 3%, 2%, or 1% death of target cells after administration of a dose less than 1 mg/kg. Reduced-cytotoxicity variants may still be cytotoxic at certain concentrations or dosages but exhibit reduced cytotoxicity, such as, e.g., are not capable of exhibiting a significant level of Shiga toxin cytotoxicity as described herein.
[0434] The ability to conjugate detection-promoting agents known in the art to various cell-targeting molecules of the present invention provides useful compositions for the detection of certain cells, such as, e.g., cancer, tumor, immune, and/or infected cells. These diagnostic embodiments of the cell-targeting molecules of the invention may be used for information gathering via various imaging techniques and assays known in the art. For example, diagnostic embodiments of the cell-targeting molecules of the invention may be used for information gathering via imaging of intracellular organelles (e.g. endocytotic, Golgi, endoplasmic reticulum, and cytosolic compartments) of individual cancer cells, immune cells, and/or infected cells in a patient or biopsy sample.
[0435] Various types of information may be gathered using the diagnostic embodiments of the cell-targeting molecules of the invention whether for diagnostic uses or other uses. This information may be useful, for example, in diagnosing neoplastic cell types, determining therapeutic susceptibilities of a patient's disease, assaying the progression of anti-neoplastic therapies over time, assaying the progression of immunomodulatory therapies over time, assaying the progression of antimicrobial therapies over time, evaluating the presence of infected cells in transplantation materials, evaluating the presence of unwanted cell types in transplantation materials, and/or evaluating the presence of residual tumor cells after surgical excision of a tumor mass.
[0436] For example, subpopulations of patients might be ascertained using information gathered using the diagnostic variants of the cell-targeting molecules of the invention, and then individual patients could be further categorized into subpopulations based on their unique characteristic(s) revealed using those diagnostic embodiments. For example, the effectiveness of specific pharmaceuticals or therapies might be a criterion used to define a patient subpopulation. For example, a nontoxic diagnostic variant of a particular cytotoxic, cell-targeting molecule of the invention may be used to differentiate which patients are in a class or subpopulation of patients predicted to respond positively to a cytotoxic variant of that cell-targeting molecule of the invention. Accordingly, associated methods for patient identification, patient stratification, and diagnosis using cell-targeting molecules of the present invention, including non-toxic variants of cytotoxic, cell-targeting molecules of the present invention, are considered to be within the scope of the present invention.
[0437] The expression of the target biomolecule by a cell need not be native in order for cell-targeting by a cell-targeting molecule of the present invention, such as, e.g., for direct cell-kill, indirect cell-kill, delivery of exogenous materials like T-cell epitopes, and/or information gathering. Cell surface expression of the target biomolecule could be the result of an infection, the presence of a pathogen, and/or the presence of an intracellular microbial pathogen. Expression of a target biomolecule could be artificial such as, for example, by forced or induced expression after infection with a viral expression vector, see e.g. adenoviral, adeno-associated viral, and retroviral systems. An example of inducing expression of a target biomolecule is the upregulation of CD38 expression of cells exposed to retinoids, like all-trans retinoic acid and various synthetic retinoids, or any retinoic acid receptor (RAR) agonist (Drach J et al., Cancer Res 54: 1746-52 (1994); Uruno A et al., J Leukoc Biol 90: 235-47 (2011)). Expression of CD30 can be induced in both B-cells and T-cells by exposure to by mitogens, phytohemagglutinin (PHA), staphylococcal protein A, EBV virus, human T-cell leukemia virus 1 or 2 (HTLV-1 or HTLV-2) (see e.g. Stein H et al., Blood 66: 848-58 (1985)). In another example, CD20, HER2, and EGFR expression may be induced by exposing a cell to ionizing radiation (Wattenberg M et al., Br J Cancer 110: 1472-80 (2014)). Further, PSMA expression is upregulated in response to androgen deprivation (see e.g. Chang S et al., Cancer 88: 407-15 (2000); Meller B et al., EJNMMI Res 5: 66 (2015)).
[0438] In certain embodiments, the molecule of the present invention is useful for tracking the behavior of an inert Shiga toxin effector polypeptide and/or cell-targeting molecule, such as, e.g., in vivo, tissue culture, and/or by a laboratory sensor. For example, a dye-conjugated Shiga toxin effector polypeptide of the present invention may be tracked using a light sensor during an experiment, e.g., both before and after an anti-Shiga toxin antibody incubation step.
IV. Pharmaceutical and Diagnostic Compositions Comprising Cell-Targeting Molecules of the Present Invention
[0439] The present invention provides Shiga toxin effector polypeptides and cell-targeting molecules for use, alone or in combination with one or more additional therapeutic agents, in a pharmaceutical composition, for treatment or prophylaxis of conditions, diseases, disorders, or symptoms described in further detail below (e.g. cancers, malignant tumors, non-malignant tumors, growth abnormalities, immune disorders, and microbial infections). The present invention further provides pharmaceutical compositions comprising a Shiga toxin polypeptide or cell-targeting molecule of the present invention, or a pharmaceutically acceptable salt or solvate thereof, according to the invention, together with at least one pharmaceutically acceptable carrier, excipient, or vehicle. In certain embodiments, the pharmaceutical composition of the present invention may comprise homo-multimeric and/or hetero-multimeric forms of a Shiga toxin effector polypeptides or cell-targeting molecule of the present invention. The pharmaceutical compositions of the invention are useful in methods of treating, ameliorating, or preventing a disease, condition, disorder, or symptom described in further detail below. Each such disease, condition, disorder, or symptom is envisioned to be a separate embodiment with respect to uses of a pharmaceutical composition according to the invention. The invention further provides pharmaceutical compositions for use in at least one method of treatment according to the invention, as described in more detail below.
[0440] As used herein, the terms patient and subject are used interchangeably to refer to any organism, commonly vertebrates such as humans and animals, which presents symptoms, signs, and/or indications of at least one disease, disorder, or condition. These terms include mammals such as the non-limiting examples of primates, livestock animals (e.g. cattle, horses, pigs, sheep, goats, etc.), companion animals (e.g. cats, dogs, etc.) and laboratory animals (e.g. mice, rabbits, rats, etc.).
[0441] As used herein, treat, treating, or treatment and grammatical variants thereof refer to an approach for obtaining beneficial or desired clinical results. The terms may refer to slowing the onset or rate of development of a condition, disorder or disease, reducing or alleviating symptoms associated with it, generating a complete or partial regression of the condition, or some combination of any of the above. For the purposes of this invention, beneficial or desired clinical results include, but are not limited to, reduction or alleviation of symptoms, diminishment of extent of disease, stabilization (e.g. not worsening) of state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. Treat, treating, or treatment can also mean prolonging survival relative to expected survival time if not receiving treatment. A subject (e.g. a human) in need of treatment may thus be a subject already afflicted with the disease or disorder in question. The terms treat, treating, or treatment includes inhibition or reduction of an increase in severity of a pathological state or symptoms relative to the absence of treatment, and is not necessarily meant to imply complete cessation of the relevant disease, disorder, or condition. With regard to tumors and/or cancers, treatment includes reduction in overall tumor burden and/or individual tumor size.
[0442] As used herein, the terms prevent, preventing, prevention and grammatical variants thereof refer to an approach for preventing the development of, or altering the pathology of, a condition, disease, or disorder. Accordingly, prevention may refer to prophylactic or preventive measures. For the purposes of this invention, beneficial or desired clinical results include, but are not limited to, prevention or slowing of symptoms, progression or development of a disease, whether detectable or undetectable. A subject (e.g. a human) in need of prevention may thus be a subject not yet afflicted with the disease or disorder in question. The term prevention includes slowing the onset of disease relative to the absence of treatment, and is not necessarily meant to imply permanent prevention of the relevant disease, disorder or condition. Thus preventing or prevention of a condition may in certain contexts refer to reducing the risk of developing the condition, or preventing or delaying the development of symptoms associated with the condition.
[0443] As used herein, an effective amount or therapeutically effective amount is an amount or dose of a composition (e.g. a therapeutic composition, compound, or agent) that produces at least one desired therapeutic effect in a subject, such as preventing or treating a target condition or beneficially alleviating a symptom associated with the condition. The most desirable therapeutically effective amount is an amount that will produce a desired efficacy of a particular treatment selected by one of skill in the art for a given subject in need thereof. This amount will vary depending upon a variety of factors understood by the skilled worker, including but not limited to the characteristics of the therapeutic composition (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including age, sex, disease type, disease stage, general physical condition, responsiveness to a given dosage, and type of medication), the nature of the pharmaceutically acceptable carrier or carriers in the formulation, and the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount through routine experimentation, namely by monitoring a subject's response to administration of a composition and adjusting the dosage accordingly (see e.g. Remington: The Science and Practice of Pharmacy (Gennaro A, ed., Mack Publishing Co., Easton, PA, U.S., 19th ed., 1995)).
[0444] Diagnostic compositions of the present invention comprise a cell-targeting molecule of the present invention and one or more detection-promoting agents. When producing or manufacturing a diagnostic composition of the present invention, a cell-targeting molecule of the present invention may be directly or indirectly linked to one or more detection-promoting agents. There are numerous standard techniques known to the skilled worker for incorporating, affixing, and/or conjugating various detection-promoting agents to proteins or proteinaceous components of molecules, especially to immunoglobulins and immunoglobulin-derived domains.
[0445] There are numerous detection-promoting agents known to the skilled worker, such as isotopes, dyes, colorimetric agents, contrast enhancing agents, fluorescent agents, bioluminescent agents, and magnetic agents, which can be operably linked to the polypeptides or cell-targeting molecules of the invention for information gathering methods, such as for diagnostic and/or prognostic applications to diseases, disorders, or conditions of an organism (see e.g. Cai W et al., J Nucl Med 48: 304-10 (2007); Nayak T, Brechbiel M, Bioconjug Chem 20: 825-41 (2009); Paudyal P et al., Oncol Rep 22: 115-9 (2009); Qiao J et al., PLoS ONE 6: e18103 (2011); Sano K et al., Breast Cancer Res 14: R61 (2012)). These agents may be associated with, linked to, and/or incorporated within the polypeptide or cell-targeting molecule of the invention at any suitable position. For example, the linkage or incorporation of the detection-promoting agent may be via an amino acid residue(s) of a molecule of the present invention or via some type of linkage known in the art, including via linkers and/or chelators. The incorporation of the agent is in such a way to enable the detection of the presence of the diagnostic composition in a screen, assay, diagnostic procedure, and/or imaging technique.
[0446] Similarly, there are numerous imaging approaches known to the skilled worker, such as non-invasive in vivo imaging techniques commonly used in the medical arena, for example: computed tomography imaging (CT scanning), optical imaging (including direct, fluorescent, and bioluminescent imaging), magnetic resonance imaging (MRI), positron emission tomography (PET), single-photon emission computed tomography (SPECT), ultrasound, and x-ray computed tomography imaging.
V. Molecules of the Present Invention Immobilized on Solid Substrates
[0447] Certain embodiments of the present invention include a molecule of the present invention (e.g. a Shiga toxin effector polypeptide, a Shiga toxin effector polypeptide scaffold, a cell-targeting molecule, fusion protein, or polynucleotide of the present invention), or any effector fragment thereof, immobilized on a solid substrate. Solid substrates contemplated herein include, but are not limited to, microbeads, nanoparticles, polymers, matrix materials, microarrays, microtiter plates, or any solid surface known in the art (see e.g. U.S. Pat. No. 7,771,955). In accordance with these embodiments, a molecule of the present invention may be covalently or non-covalently linked to a solid substrate, such as, e.g., a bead, particle, or plate, using techniques known to the skilled worker (see e.g. Jung Y et al., Analyst 133: 697-701 (2008)). Immobilized molecules of the invention may be used for screening applications using techniques known in the art (see e.g. Bradbury A et al., Nat Biotechnol 29: 245-54 (2011); Sutton C, Br J Pharmacol 166: 457-75 (2012); Diamante L et al., Protein Eng Des Sel 26: 713-24 (2013); Houlihan G et. al., J Immunol Methods 405: 47-56 (2014)).
[0448] Non-limiting examples of solid substrates to which a molecule of the invention may be immobilized on include: microbeads, nanoparticles, polymers, nanopolymers, nanotubes, magnetic beads, paramagnetic beads, superparamagnetic beads, streptavidin coated beads, reverse-phase magnetic beads, carboxy terminated beads, hydrazine terminated beads, silica (sodium silica) beads and iminodiacetic acid (IDA)-modified beads, aldehyde-modified beads, epoxy-activated beads, diaminodipropylamine (DADPA)-modified beads (beads with primary amine surface group), biodegradable polymeric beads, polystyrene substrates, amino-polystyrene particles, carboxyl-polystyrene particles, epoxy-polystyrene particles, dimethylamino-polystyrene particles, hydroxy-polystyrene particles, colored particles, flow cytometry particles, sulfonate-polystyrene particles, nitrocellulose surfaces, reinforced nitrocellulose membranes, nylon membranes, glass surfaces, activated glass surfaces, activated quartz surfaces, polyvinylidene difluoride (PVDF) membranes, polyacrylamide-based substrates, poly-vinyl chloride substrates, poly-methyl methacrylate substrates, poly(dimethyl siloxane) substrates, and photopolymers which contain photoreactive species (such as nitrenes, carbenes, and ketyl radicals) capable of forming covalent linkages. Other examples of solid substrates to which a molecule of the invention may be immobilized on are commonly used in molecular display systems, such as, e.g., cellular surfaces, phages, and virus particles.
VI. Production or Manufacture of Pharmaceutical and/or Diagnostic Compositions Comprising Cell-Targeting Molecules of the Present Invention
[0449] Pharmaceutically acceptable salts or solvates of any of the Shiga toxin effector polypeptides, Shiga toxin effector polypeptide scaffolds, and cell-targeting molecules of the present invention are within the scope of the present invention. In addition, pharmaceutical compositions comprising a salt or solvate of a cell-targeting molecule(s) of the present invention are within the scope of the present invention.
[0450] The term solvate in the context of the present invention refers to a complex of defined stoichiometry formed between a solute (in casu, a proteinaceous compound or pharmaceutically acceptable salt thereof according to the invention) and a solvent. The solvent in this connection may, for example, be water, ethanol or another pharmaceutically acceptable, typically small-molecular organic species, such as, but not limited to, acetic acid or lactic acid. When the solvent in question is water, such a solvate is normally referred to as a hydrate.
[0451] Polypeptides and proteins of the present invention, or salts thereof, may be formulated as pharmaceutical compositions prepared for storage or administration, which typically comprise a therapeutically effective amount of a molecule of the present invention, or a salt thereof, in a pharmaceutically acceptable carrier. The term pharmaceutically acceptable carrier includes any of the standard pharmaceutical carriers. Pharmaceutically acceptable carriers for therapeutic molecule use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences (Mack Publishing Co. (A. Gennaro, ed., 1985). As used herein, pharmaceutically acceptable carrier includes any and all physiologically acceptable, i.e. compatible, solvents, dispersion media, coatings, antimicrobial agents, isotonic, and absorption delaying agents, and the like. Pharmaceutically acceptable carriers or diluents include those used in formulations suitable for oral, rectal, nasal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, and transdermal) administration. Exemplary pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyloleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. In certain embodiments, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g. by injection or infusion). Depending on selected route of administration, the protein or other pharmaceutical component may be coated in a material intended to protect the compound from the action of low pH and other natural inactivating conditions to which the active protein may encounter when administered to a patient by a particular route of administration.
[0452] Therapeutic compositions of the present invention are typically sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, solvate, salt, powder, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier may be a solvent or dispersion medium containing, for example, water, alcohol such as ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), or any suitable mixture. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by use of surfactants according to formulation chemistry well known in the art. In certain embodiments, the pharmaceutical composition of the present invention may comprise one or more isotonic agents, such as, e.g., a sugar, polyalcohol, and/or ions like mannitol, sorbitol, and sodium chloride
[0453] A pharmaceutical composition of the present invention optionally includes a pharmaceutically acceptable excipient. Non-limiting examples of pharmaceutically acceptable excipients include arginine, arginine sulfate, glycerol, mannitol, methionine, polysorbate, sodium chloride, sorbitol, sucrose, and/or trehalose. In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier and at least one pharmaceutically acceptable excipient. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising at least one pharmaceutically acceptable excipient. In certain embodiments of the pharmaceutical composition of the present invention, the excipient functions to reduce and/or limit the immunogenicity and/or immunogenic potential of the cell-targeting molecule, such as, e.g. after administration and/or repeated administration to a mammal.
[0454] The pharmaceutical compositions of the present invention may comprise one or more adjuvants such as a buffer, tonicity-adjusting agent (isotonic agent), antioxidant, surfactant, stabilizer, preservative, emulsifying agent, cryoprotective agent, wetting agent, and/or dispersing agent or other additives well known to those of skill in the art, such as, e.g. a binding agent. In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier and a pharmaceutically acceptable adjuvant or other additive. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising a pharmaceutically acceptable adjuvant or other additive. A non-limiting example of a pharmaceutically suitable stabilizer is human albumin.
[0455] The pharmaceutical composition of the present invention may comprise one or more pharmaceutically acceptable buffers. Non-limiting examples of suitable buffers include acetate, citrate, histidine, phosphate, and succinate buffers. In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier comprising a pharmaceutically acceptable buffer. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising a pharmaceutically acceptable buffer.
[0456] The pharmaceutical composition of the present invention may comprise one or more pharmaceutically acceptable isotonic agents or tonicity-adjusting agents. Non-limiting examples of suitable isotonic agents include sugars (e.g. dextrose), sugar alcohols, sodium chloride, and the like. Further examples of suitable sugars include disaccharides like sucrose and trehalose. Exemplary, pharmaceutically acceptable sugar alcohols include glycerol, mannitol, and sorbitol. In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier and a pharmaceutically acceptable isotonic agent. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising a pharmaceutically acceptable isotonic agent.
[0457] The pharmaceutical compositions of the present invention may comprise one or more pharmaceutically acceptable antioxidants. Exemplary pharmaceutically acceptable antioxidants include water soluble antioxidants, such as, e.g., ascorbic acid, cysteine hydrochloride, methionine, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as, e.g., ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and metal-chelating agents, such as, e.g., citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like. In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier and a pharmaceutically acceptable antioxidant. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising a pharmaceutically acceptable antioxidant.
[0458] A pharmaceutical composition of the present invention may comprise one or more pharmaceutically acceptable surfactants and/or emulsifying agents (emulsifiers). Non-limiting examples of suitable surfactants and/or emulsifiers include polysorbates such as, e.g., polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan monopalmitate (polysorbate 40), polyoxyethylene (20) sorbitan monostearate (polysorbate 60), and (polyoxyethylene (20) sorbitan monooleate (polysorbate 80). In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier and a pharmaceutically acceptable surfactant and/or emulsifier. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising a pharmaceutically acceptable surfactant and/or emulsifier. One or more surfactants and/or emulsifying agents may also be desirable in a pharmaceutical composition of the present invention to help prevent aggregation of the cell-targeting molecule of the present invention.
[0459] The pharmaceutical compositions of the present invention may comprise one or more pharmaceutically acceptable preservative agents. For example, preventing the presence of microorganisms may be ensured both by sterilization procedures, and by the inclusion of various antibacterial and antifungal agents, such as, e.g., paraben, chlorobutanol, phenol, sorbic acid, and the like in the compositions of the present invention.
[0460] A pharmaceutical composition of the present invention may comprise one or more pharmaceutically acceptable cryoprotective agents (cryoprotectants). Non-limiting examples of suitable cryoprotectants include ethylene glycol, glycerol, sucrose, and trehalose. In certain embodiments, the pharmaceutical composition of the present invention comprises an aqueous carrier and a pharmaceutically acceptable cryoprotectant. In certain other embodiments, the pharmaceutical composition of the present invention comprises a salt and/or powder, such as, e.g. a freeze-dried, lyophilized, dehydrated, and/or cryodessicated composition comprising a pharmaceutically acceptable cryoprotectant.
[0461] In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as, e.g, a monostearate salt, aluminum monostearate, and/or gelatin.
[0462] In another aspect, the present invention provides pharmaceutical compositions comprising one or a combination of different polypeptides and/or cell-targeting molecules of the invention, or an ester, salt or amide of any of the foregoing, and at least one pharmaceutically acceptable carrier.
[0463] The pH of the pharmaceutical composition of the present invention can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide, or buffers with acetate, citrate, histidine, succinate, phosphate, and the like. Non-limiting examples of pharmaceutically acceptable solvents or carriers for use in a pharmaceutical composition of the present invention include aqueous solutions comprising a cell-targeting molecule of the present invention and a buffer such as, e.g., citrate, histidine, phosphate, or succinate adjusted to pH 5.0, 6.0, 7.0, or 4.0, respectively. Certain embodiments of the present invention include compositions comprising one of the aforementioned solvents and/or carriers of the present invention.
[0464] Pharmaceutical compositions of the present invention that are solutions or suspensions used for intradermal or subcutaneous application typically include one or more of: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid, cysteine hydrochloride, methionine, sodium bisulfate, sodium metabisulfite, and sodium sulfite; chelating agents such as citric acid, ethylenediaminetetraacetic acid, sorbitol, tartaric acid, and phosphoric acid; surfactants such as a polysorbate; buffers such as acetate, citrate, histidine, and phosphate buffers; and tonicity adjusting agents such as, e.g., dextrose, glycerol, mannitol, sodium chloride, sorbitol, sucrose, and trehalose. Such preparations may be enclosed in ampoules, disposable syringes or multiple dose vials made of a glass or plastic.
[0465] Sterile injectable solutions may be prepared by incorporating a protein or cell-targeting molecule of the present invention in the required amount in an appropriate solvent with one or a combination of ingredients described above, as required, followed by sterilization microfiltration. Dispersions may be prepared by incorporating the active compound into a sterile vehicle that contains a dispersion medium and other ingredients, such as those described above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient in addition to any additional desired ingredient from a sterile-filtered solution thereof. In certain embodiments, the pharmaceutical composition of the present invention comprises a powder comprising sorbitol, trehalose, sodium citrate, and polysorbate-20, and optionally, further comprises glycerol and/or methionine. In certain embodiments, the pharmaceutical composition of the present invention comprises sodium citrate, trehalose, and polysorbate-20, and optionally, further comprises glycerol and/or methionine.
[0466] When a therapeutically effective amount of a polypeptide and/or cell-targeting molecule of the invention is designed to be administered by, e.g. intravenous, cutaneous or subcutaneous injection, the binding agent will be in the form of a pyrogen-free, parenterally acceptable aqueous solution. Methods for preparing parenterally acceptable protein solutions, taking into consideration appropriate pH, isotonicity, stability, and the like, are within the skill in the art. A preferred pharmaceutical composition for intravenous, cutaneous, or subcutaneous injection will contain, in addition to binding agents, an isotonic vehicle such as sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection, lactated Ringer's injection, or other vehicle as known in the art.
[0467] In certain embodiments, the pharmaceutical composition of the present invention comprises sorbitol, sodium citrate, and polysorbate-20, and optionally, further comprises albumin, glycerol, and/or methionine. In certain embodiments, the pharmaceutical composition of the present invention comprises sorbitol, histidine, and polysorbate-20, and optionally, further comprises albumin, glycerol, and/or methionine.
[0468] The formulations of the pharmaceutical compositions of the invention may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. In such form, the composition is divided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparations, for example, packeted tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms. It may be provided in single dose injectable form, for example in the form of an autoinjector or pen. Compositions of the present invention may be formulated for any suitable route and means of administration. Subcutaneous or transdermal modes of administration may be particularly suitable for therapeutic molecules described herein.
[0469] As described elsewhere herein, a polypeptide and/or cell-targeting molecule of the present invention may be prepared with carriers that will protect the active therapeutic agent against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art (see e.g. Sustained and Controlled Release Drug Delivery Systems (Robinson J, ed., Marcel Dekker, Inc., NY, U.S., 1978)).
[0470] In certain embodiments, the composition of the present invention (e.g. a pharmaceutical and/or diagnostic composition) may be formulated to ensure a desired in vivo distribution of a cell-targeting molecule of the present invention. For example, the blood-brain barrier excludes many large and/or hydrophilic compounds. To target a therapeutic molecule or composition of the present invention to a particular in vivo location, they can be formulated, for example, in liposomes which may comprise one or more moieties that are selectively transported into specific cells or organs, thus enhancing targeted drug delivery. Exemplary targeting moieties include folate or biotin, mannosides, antibodies, surfactant protein A receptor, p120 catenin, and the like.
[0471] Pharmaceutical compositions include parenteral formulations designed to be used as implants or particulate systems. Examples of implants are depot formulations composed of polymeric or hydrophobic components such as emulsions, ion exchange resins, and soluble salt solutions. Examples of particulate systems are microspheres, microparticles, nanocapsules, nanospheres, and nanoparticles (see e.g. Honda M et al., Int J Nanomedicine 8: 495-503 (2013); Sharma A et al., Biomed Res Int 2013: 960821 (2013); Ramishetti S, Huang L, Ther Deliv 3: 1429-45 (2012)). Controlled release formulations may be prepared using polymers sensitive to ions, such as, e.g. liposomes, polaxamer 407, and hydroxyapatite.
VII. Polynucleotides, Expression Vectors, and Host Cells of the Present Invention
[0472] Beyond the polypeptides and cell-targeting molecules of the present invention, the polynucleotides that encode the polypeptides, proteins, and cell-targeting molecules of the invention, or functional portions thereof, are also encompassed within the scope of the present invention. The term polynucleotide is equivalent to the term nucleic acid, each of which includes one or more of: polymers of deoxyribonucleic acids (DNAs), polymers of ribonucleic acids (RNAs), analogs of these DNAs or RNAs generated using nucleotide analogs, and derivatives, fragments and homologs thereof. The polynucleotide of the present invention may be single-, double-, or triple-stranded. Such polynucleotides are specifically disclosed to include all polynucleotides capable of encoding an exemplary protein, for example, taking into account the wobble known to be tolerated in the third position of RNA codons, yet encoding for the same amino acid as a different RNA codon (see Stothard P, Biotechniques 28: 1102-4 (2000)).
[0473] In one aspect, the present invention provides polynucleotides which encode a Shiga toxin effector polypeptide and/or cell-targeting molecule of the present invention, or a fragment or derivative thereof. The polynucleotides may include, e.g., a nucleic acid sequence encoding a polypeptide at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or more, identical to a polypeptide comprising one of the amino acid sequences of a polypeptide or cell-targeting molecule of the present invention. The invention also includes polynucleotides comprising nucleotide sequences that hybridize under stringent conditions to a polynucleotide which encodes Shiga toxin effector polypeptide and/or cell-targeting molecule of the invention, or a fragment or derivative thereof, or the antisense or complement of any such sequence.
[0474] Derivatives or analogs of the molecules of the present invention (e.g., Shiga toxin effector polypeptides of the present invention and cell-targeting molecules comprising the same) include, inter alia, polynucleotide (or polypeptide) molecules having regions that are substantially homologous to the polynucleotides (or Shiga toxin effector polypeptides and cell-targeting molecules of the present invention), e.g. by at least about 45%, 50%, 70%, 80%, 95%, 98%, or even 99% identity (with a preferred identity of 80-99%) over a polynucleotide (or polypeptide) sequence of the same size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art. An exemplary program is the GAP program (Wisconsin Sequence Analysis Package, Version 8 for UNIX, Genetics Computer Group, University Research Park, Madison, WI, U.S.) using the default settings, which uses the algorithm of Smith T, Waterman M, Adv Appl Math 2: 482-9 (1981). Also included are polynucleotides capable of hybridizing to the complement of a sequence encoding the cell-targeting proteins of the invention under stringent conditions (see e.g. Ausubel F et al., Current Protocols in Molecular Biology (John Wiley & Sons, New York, NY, U.S., 1993)), and below. Stringent conditions are known to those skilled in the art and may be found, e.g., in Current Protocols in Molecular Biology (John Wiley & Sons, NY, U.S., Ch. Sec. 6.3.1-6.3.6 (1989)).
[0475] The present invention further provides expression vectors that comprise the polynucleotides within the scope of the present invention. The polynucleotides capable of encoding the Shiga toxin effector polypeptides and/or cell-targeting molecules of the invention may be inserted into known vectors, including bacterial plasmids, viral vectors and phage vectors, using material and methods well known in the art to produce expression vectors. Such expression vectors will include the polynucleotides necessary to support production of contemplated Shiga toxin effector polypeptides and/or cell-targeting molecules of the invention within any host cell of choice or cell-free expression systems (e.g. pTxb1 and pIVEX2.3). The specific polynucleotides comprising expression vectors for use with specific types of host cells or cell-free expression systems are well known to one of ordinary skill in the art, can be determined using routine experimentation, and/or may be purchased.
[0476] The term expression vector, as used herein, refers to a polynucleotide, linear or circular, comprising one or more expression units. The term expression unit denotes a polynucleotide segment encoding a polypeptide of interest and capable of providing expression of the nucleic acid segment in a host cell. An expression unit typically comprises a transcription promoter, an open reading frame encoding the polypeptide of interest, and a transcription terminator, all in operable configuration. An expression vector contains one or more expression units. Thus, in the context of the present invention, an expression vector encoding a Shiga toxin effector polypeptide and/or cell-targeting molecule of the invention comprising a single polypeptide chain includes at least an expression unit for the single polypeptide chain, whereas a protein comprising, e.g. two or more polypeptide chains (e.g. one chain comprising a V.sub.L domain and a second chain comprising a V.sub.H domain linked to a toxin effector polypeptide) includes at least two expression units, one for each of the two polypeptide chains of the protein. For expression of multi-chain cell-targeting proteins of the invention, an expression unit for each polypeptide chain may also be separately contained on different expression vectors (e.g. expression may be achieved with a single host cell into which expression vectors for each polypeptide chain has been introduced).
[0477] Expression vectors capable of directing transient or stable expression of polypeptides and proteins are well known in the art. The expression vectors generally include, but are not limited to, one or more of the following: a heterologous signal sequence or peptide, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence, each of which is well known in the art. Optional regulatory control sequences, integration sequences, and useful markers that can be employed are known in the art.
[0478] The term host cell refers to a cell which can support the replication or expression of the expression vector. Host cells may be prokaryotic cells, such as E. coli or eukaryotic cells (e.g. yeast, insect, amphibian, bird, or mammalian cells). Creation and isolation of host cell lines comprising a polynucleotide of the invention or capable of producing a polypeptide and/or cell-targeting molecule of the present invention can be accomplished using standard techniques known in the art.
[0479] Shiga toxin effector polypeptides and/or proteins within the scope of the present invention may be variants or derivatives of the polypeptides and molecules described herein that are produced by modifying the polynucleotide encoding a polypeptide and/or proteinaceous component of a cell-targeting molecule by altering one or more amino acids or deleting or inserting one or more amino acids that may render it more suitable to achieve desired properties, such as more optimal expression by a host cell.
VIII. Delivery Devices and Kits
[0480] In certain embodiments, the invention relates to a device comprising one or more compositions of matter of the present invention, such as a pharmaceutical composition or diagnostic composition, for delivery to a subject in need thereof. Thus, a delivery device comprising one or more compositions of the present invention can be used to administer to a patient a composition of matter of the present invention by various delivery methods, including: intravenous, subcutaneous, intramuscular or intraperitoneal injection; oral administration; transdermal administration; pulmonary or transmucosal administration; administration by implant, osmotic pump, cartridge or micro pump; or by other means recognized by a person of skill in the art.
[0481] Also within the scope of the present invention are kits comprising at least one composition of matter of the invention, and optionally, packaging and instructions for use. Kits may be useful for drug administration and/or diagnostic information gathering. A kit of the invention may optionally comprise at least one additional reagent (e.g., standards, markers and the like). Kits typically include a label indicating the intended use of the contents of the kit. The kit may further comprise reagents and other tools for detecting a cell type (e.g. a tumor cell) in a sample or in a subject, or for diagnosing whether a patient belongs to a group that responds to a therapeutic strategy which makes use of a compound, composition, or related method of the present invention, e.g., such as a method described herein.
IX. Methods for Using Cell-Targeting Molecules of the Present Invention and/or Pharmaceutical and/or Diagnostic Compositions Thereof
[0482] Generally, it is an object of the present invention to provide pharmacologically active agents, as well as compositions comprising the same, that can be used in the prevention and/or treatment of diseases, disorders, and conditions, such as certain cancers, tumors, growth abnormalities, immune disorders, or further pathological conditions mentioned herein. Accordingly, the present invention provides methods of using the polypeptides, cell-targeting molecules, and pharmaceutical compositions of the invention for the targeted killing of cells, for delivering additional exogenous materials into targeted cells, for labeling of the interiors of targeted cells, for collecting diagnostic information, for the delivering of T-cell epitopes to the MHC class I presentation pathway of target cells, and for treating diseases, disorders, and conditions as described herein. For example, the methods of the present invention may be used to prevent or treat cancers, cancer initiation, tumor initiation, metastasis, and/or disease reoccurrence.
[0483] In particular, it is an object of the invention to provide such pharmacologically active agents, compositions, and/or methods that have certain advantages compared to the agents, compositions, and/or methods that are currently known in the art. Accordingly, the present invention provides methods of using Shiga toxin effector polypeptides and cell-targeting molecules with specified protein sequences and pharmaceutical compositions thereof. For example, any of the amino acid sequences in SEQ ID NOs: 4-1140 may be specifically utilized as a component of the cell-targeting molecule used in the following methods or any method for using a cell-targeting molecule known to the skilled worker, such as, e.g., various methods described in WO 2014/164680, WO 2014/164693, WO 2015/138435, WO 2015/138452, WO 2015/113005; WO 2015/113007, WO 2015/138435, WO 2015/138452, US20150259428, WO 2015/191764, US20160177284, and WO 2016/126950.
[0484] The present invention provides methods of killing a cell comprising the step of contacting the cell, either in vitro or in vivo, with a Shiga toxin effector polypeptide, cell-targeting molecule, or pharmaceutical composition of the present invention. The Shiga toxin effector polypeptides, cell-targeting molecules, and pharmaceutical compositions of the present invention can be used to kill a specific cell type upon contacting a cell or cells with one of the claimed compositions of matter. In certain embodiments, a cell-targeting molecule or pharmaceutical composition of the present invention can be used to kill specific cell types in a mixture of different cell types, such as mixtures comprising cancer cells, infected cells, and/or hematological cells. In certain embodiments, a cell-targeting molecule, or pharmaceutical composition of the present invention can be used to kill cancer cells in a mixture of different cell types. In certain embodiments, a cytotoxic Shiga cell-targeting molecule, or pharmaceutical composition of the present invention can be used to kill specific cell types in a mixture of different cell types, such as pre-transplantation tissues. In certain embodiments, a Shiga toxin effector polypeptide, cell-targeting molecule, or pharmaceutical composition of the present invention can be used to kill specific cell types in a mixture of cell types, such as pre-administration tissue material for therapeutic purposes. In certain embodiments, a cell-targeting molecule or pharmaceutical composition of the present invention can be used to selectively kill cells infected by viruses or microorganisms, or otherwise selectively kill cells expressing a particular extracellular target biomolecule, such as a cell surface biomolecule. The Shiga toxin effector polypeptides, cell-targeting molecules, and pharmaceutical compositions of the present invention have varied applications, including, e.g., uses in depleting unwanted cell types from tissues either in vitro or in vivo, uses in modulating immune responses to treat graft versus host, uses as antiviral agents, uses as anti-parasitic agents, and uses in purging transplantation tissues of unwanted cell types.
[0485] In certain embodiments, certain Shiga toxin effector polypeptides, cell-targeting molecules, and pharmaceutical compositions of the present invention, alone or in combination with other compounds or pharmaceutical compositions, can show potent cell-kill activity when administered to a population of cells, in vitro or in vivo in a subject such as in a patient in need of treatment. By targeting the delivery of enzymatically active Shiga toxin A Subunit effector polypeptides and/or T-cell epitopes using high-affinity binding regions to specific cell types, cell-kill activities can be restricted to specifically and selectively killing certain cell types within an organism, such as certain cancer cells, neoplastic cells, malignant cells, non-malignant tumor cells, and/or infected cells.
[0486] The present invention provides a method of killing a cell in a patient in need thereof, the method comprising the step of administering to the patient at least one cell-targeting molecule of the present invention or a pharmaceutical composition thereof.
[0487] In certain embodiments, the cell-targeting molecule of the present invention or pharmaceutical compositions thereof can be used to kill a cancer cell in a patient by targeting an extracellular biomolecule found physically coupled with a cancer or tumor cell. The terms cancer cell or cancerous cell refers to various neoplastic cells which grow and divide in an abnormally accelerated and/or unregulated fashion and will be clear to the skilled person. The term tumor cell includes both malignant and non-malignant cells. Generally, cancers and/or tumors can be defined as diseases, disorders, or conditions that are amenable to treatment and/or prevention. The cancers and tumors (either malignant or non-malignant) which are comprised of cancer cells and/or tumor cells which may benefit from methods and compositions of the invention will be clear to the skilled person. Neoplastic cells are often associated with one or more of the following: unregulated growth, lack of differentiation, local tissue invasion, angiogenesis, and metastasis. The diseases, disorders, and conditions resulting from cancers and/or tumors (either malignant or non-malignant) which may benefit from the methods and compositions of the present invention targeting certain cancer cells and/or tumor cells will be clear to the skilled person.
[0488] Certain embodiments of the cell-targeting molecules and compositions of the present invention may be used to kill cancer stem cells, tumor stem cells, pre-malignant cancer-initiating cells, and tumor-initiating cells, which commonly are slow dividing and resistant to cancer therapies like chemotherapy and radiation. For example, acute myeloid leukemias (AMLs) may be treated with the present invention by killing AML stem cells and/or dormant AML progenitor cells (see e.g. Shlush L et al., Blood 120: 603-12 (2012)). Cancer stem cells often overexpress cell surface targets, such as, e.g., CD44, CD200, and others listed herein, which can be targets of certain binding regions of certain embodiments of the cell-targeting molecules of the present invention (see e.g. Kawasaki B et al., Biochem Biophys Res Commun 364:778-82 (2007); Reim F et al., Cancer Res 69: 8058-66 (2009)).
[0489] Because of the Shiga toxin A Subunit based mechanism of action, compositions of matter of the present invention may be more effectively used in methods involving their combination with, or in complementary fashion with other therapies, such as, e.g., chemotherapies, immunotherapies, radiation, stem cell transplantation, and immune checkpoint inhibitors, and/or effective against chemoresistant/radiation-resistant and/or resting tumor cells/tumor initiating cells/stem cells. Similarly, compositions of matter of the present invention may be more effectively used in methods involving in combination with other cell-targeted therapies targeting other than the same epitope on, non-overlapping, or different targets for the same disease disorder or condition.
[0490] Certain embodiments of the cell-targeting molecules of the present invention, or pharmaceutical compositions thereof, can be used to kill an immune cell (whether healthy or malignant) in a patient by targeting an extracellular biomolecule found physically coupled with an immune cell.
[0491] It is within the scope of the present invention to utilize a cell-targeting molecule of the present invention, or pharmaceutical composition thereof, for the purposes of purging patient cell populations (e.g. bone marrow) of malignant, neoplastic, or otherwise unwanted T-cells and/or B-cells and then reinfusing the T-cell and/or B-cells depleted material into the patient (see e.g. van Heeckeren W et al., Br J Haematol 132: 42-55 (2006); (see e.g. Alpdogan O, van den Brink M, Semin Oncol 39: 629-42 (2012)).
[0492] It is within the scope of the present invention to utilize the cell-targeting molecule of the present invention, or pharmaceutical composition thereof, for the purposes of ex vivo depletion of T cells and/or B-cells from isolated cell populations removed from a patient. In one non-limiting example, the cell-targeting molecule of the invention can be used in a method for prophylaxis of organ and/or tissue transplant rejection wherein the donor organ or tissue is perfused prior to transplant with a cytotoxic, cell-targeting molecule of the invention or a pharmaceutical composition thereof in order to purge the organ of donor T-cells and/or B-cells (see e.g. Alpdogan O, van den Brink M, Semin Oncol 39: 629-42 (2012)).
[0493] It is also within the scope of the present invention to utilize the cell-targeting molecule of the invention, or pharmaceutical composition thereof, for the purposes of depleting T-cells and/or B-cells from a donor cell population as a prophylaxis against graft-versus-host disease, and induction of tolerance, in a patient to undergo a bone marrow and or stem cell transplant (see e.g. van Heeckeren W et al., Br J Haematol 132: 42-55 (2006); (see e.g. Alpdogan O, van den Brink M, Semin Oncol 39: 629-42 (2012)).
[0494] In certain embodiments of the Shiga toxin effector polypeptide or cell-targeting molecule of the present invention, or pharmaceutical compositions thereof, can be used to kill an infected cell in a patient by targeting an extracellular biomolecule found physically coupled with an infected cell.
[0495] In certain embodiments of the cell-targeting molecules of the present invention, or pharmaceutical compositions thereof, can be used to seed a locus within a chordate with non-self, T-cell epitope-peptide presenting cells in order to activate the immune system to enhance policing of the locus. In certain further embodiments of this seeding method of the present invention, the locus is a tumor mass or infected tissue site. In preferred embodiments of this seeding method of the present invention, the non-self, T-cell epitope-peptide is selected from the group consisting of: peptides not already presented by the target cells of the cell-targeting molecule, peptides not present within any protein expressed by the target cell, peptides not present within the proteome or transcriptome of the target cell, peptides not present in the extracellular microenvironment of the site to be seeded, and peptides not present in the tumor mass or infect tissue site to be targeting.
[0496] This seeding method functions to label one or more target cells within a chordate with one or more MHC class I presented T-cell epitopes for recognition by effector T-cells and activation of downstream immune responses. By exploiting the cell internalizing, intracellularly routing, and T-cell epitope delivering functions of the cell-targeting molecules of the present invention, the target cells which display the delivered T-cell epitope are harnessed to induce recognition of the presenting target cell by host T-cells and induction of further immune responses including target-cell-killing by CTLs. This seeding method of using a cell-targeting molecule of the present invention can provide a temporary vaccination-effect by inducing adaptive immune responses to attack the cells within the seeded microenvironment, such as, e.g. a tumor mass or infected tissue site, whether presenting a cell-targeting molecule-delivered T-cell epitope(s) or not. This seeding method may also induce the breaking of immuno-tolerance to a target cell population, a tumor mass, and/or infected tissue site within a chordate.
[0497] Certain methods of the present invention involving the seeding of a locus within a chordate with one or more antigenic and/or immunogenic epitopes may be combined with the administration of immunologic adjuvants, whether administered locally or systemically, to stimulate the immune response to certain antigens, such as, e.g., the co-administration of a composition of the present invention with one or more immunologic adjuvants like a cytokine, bacterial product, or plant saponin. Other examples of immunologic adjuvants which may be suitable for use in the methods of the present invention include aluminum salts and oils, such as, e.g., alums, aluminum hydroxide, mineral oils, squalene, paraffin oils, peanut oils, and thimerosal.
[0498] Additionally, the present invention provides a method of treating a disease, disorder, or condition in a patient comprising the step of administering to a patient in need thereof a therapeutically effective amount of at least one of the cell-targeting molecules of the present invention, or a pharmaceutical composition thereof. Contemplated diseases, disorders, and conditions that can be treated using this method include cancers, malignant tumors, non-malignant tumors, growth abnormalities, immune disorders, and microbial infections. Administration of a therapeutically effective dosage of a composition of the present invention can result in a decrease in severity of disease symptoms, an increase in frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to the disease affliction.
[0499] The therapeutically effective amount of a composition of the present invention will depend on the route of administration, the type of organism being treated, and the physical characteristics of the specific patient under consideration. These factors and their relationship to determining this amount are well known to skilled practitioners in the medical arts. This amount and the method of administration can be tailored to achieve optimal efficacy, and may depend on such factors as weight, diet, concurrent medication and other factors, well known to those skilled in the medical arts. The dosage sizes and dosing regimen most appropriate for human use may be guided by the results obtained by the present invention, and may be confirmed in properly designed clinical trials. An effective dosage and treatment protocol may be determined by conventional means, starting with a low dose in laboratory animals and then increasing the dosage while monitoring the effects, and systematically varying the dosage regimen as well. Numerous factors may be taken into consideration by a clinician when determining an optimal dosage for a given subject. Such considerations are known to the skilled person.
[0500] An acceptable route of administration may refer to any administration pathway known in the art, including but not limited to aerosol, enteral, nasal, ophthalmic, oral, parenteral, rectal, vaginal, or transdermal (e.g. topical administration of a cream, gel or ointment, or by means of a transdermal patch). Parenteral administration is typically associated with injection at or in communication with the intended site of action, including infraorbital, infusion, intraarterial, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrathecal, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal administration.
[0501] For administration of a pharmaceutical composition of the present invention, the dosage range will generally be from about 0.001 to 10 milligrams per kilogram (mg/kg), and more, usually 0.001 to 0.5 mg/kg, of the subject's body weight. Exemplary dosages may be 0.01 mg/kg body weight, 0.03 mg/kg body weight, 0.07 mg/kg body weight, 0.9 mg/kg body weight or 0.1 mg/kg body weight or within the range of 0.01 to 0.1 mg/kg. An exemplary treatment regime is a once or twice daily administration, or a once or twice weekly administration, once every two weeks, once every three weeks, once every four weeks, once a month, once every two or three months or once every three to 6 months. Dosages may be selected and readjusted by the skilled health care professional as required to maximize therapeutic benefit for a particular patient.
[0502] Pharmaceutical compositions of the present invention will typically be administered to the same patient on multiple occasions. Intervals between single dosages can be, for example, two to five days, weekly, monthly, every two or three months, every six months, or yearly. Intervals between administrations can also be irregular, based on regulating blood levels or other markers in the subject or patient. Dosage regimens for a composition of the present invention include intravenous administration of 1 mg/kg body weight or 3 mg/kg body weight with the composition administered every two to four weeks for six dosages, then every three months at 3 mg/kg body weight or 1 mg/kg body weight.
[0503] A pharmaceutical composition of the present invention may be administered via one or more routes of administration, using one or more of a variety of methods known in the art. As will be appreciated by the skilled worker, the route and/or mode of administration will vary depending upon the desired results. Routes of administration for cell-targeting molecules and pharmaceutical compositions of the present invention include, e.g. intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal, or other parenteral routes of administration, for example by injection or infusion. For other embodiments, a cell-targeting molecule or pharmaceutical composition of the invention may be administered by a non-parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually, or topically.
[0504] Therapeutic cell-targeting molecules or pharmaceutical compositions of the present invention may be administered with one or more of a variety of medical devices known in the art. For example, in one embodiment, a pharmaceutical composition of the invention may be administered with a needleless hypodermic injection device. Examples of well-known implants and modules useful in the present invention are in the art, including e.g., implantable micro-infusion pumps for controlled rate delivery; devices for administering through the skin; infusion pumps for delivery at a precise infusion rate; variable flow implantable infusion devices for continuous drug delivery; and osmotic drug delivery systems. These and other such implants, delivery systems, and modules are known to those skilled in the art.
[0505] The cell-targeting molecule or pharmaceutical composition of the present invention may be administered alone or in combination with one or more other therapeutic or diagnostic agents. A combination therapy may include a cell-targeting molecule of the present invention, or pharmaceutical composition thereof, combined with at least one other therapeutic agent selected based on the particular patient, disease or condition to be treated. Examples of other such agents include, inter alia, a cytotoxic, anti-cancer or chemotherapeutic agent, an anti-inflammatory or anti-proliferative agent, an antimicrobial or antiviral agent, growth factors, cytokines, an analgesic, a therapeutically active small molecule or polypeptide, a single chain antibody, a classical antibody or fragment thereof, or a nucleic acid molecule which modulates one or more signaling pathways, and similar modulating therapeutic molecules which may complement or otherwise be beneficial in a therapeutic or prophylactic treatment regimen.
[0506] Treatment of a patient with cell-targeting molecule or pharmaceutical composition of the present invention preferably leads to cell death of targeted cells and/or the inhibition of growth of targeted cells. As such, cytotoxic, cell-targeting molecules of the present invention, and pharmaceutical compositions comprising them, will be useful in methods for treating a variety of pathological disorders in which killing or depleting target cells may be beneficial, such as, inter alia, cancer, tumors, other growth abnormalities, immune disorders, and infected cells. The present invention provides methods for suppressing cell proliferation, and treating cell disorders, including neoplasia, overactive B-cells, and overactive T-cells.
[0507] In certain embodiments, the cell-targeting molecules and pharmaceutical compositions of the present invention can be used to treat or prevent cancers, tumors (malignant and non-malignant), growth abnormalities, immune disorders, and microbial infections. In a further aspect, the above ex vivo method can be combined with the above in vivo method to provide methods of treating or preventing rejection in bone marrow transplant recipients, and for achieving immunological tolerance.
[0508] In certain embodiments, the present invention provides methods for treating malignancies or neoplasms and other blood cell associated cancers in a mammalian subject, such as a human, the method comprising the step of administering to a subject in need thereof a therapeutically effective amount of a cytotoxic cell-targeting molecule or pharmaceutical composition of the present invention.
[0509] The cell-targeting molecules and pharmaceutical compositions of the present invention have varied applications, including, e.g., uses in removing unwanted T-cells, uses in modulating immune responses to treat graft versus host, uses as antiviral agents, uses as antimicrobial agents, and uses in purging transplantation tissues of unwanted cell types. The cell-targeting molecules and pharmaceutical compositions of the present invention are commonly anti-neoplastic agentsmeaning they are capable of treating and/or preventing the development, maturation, or spread of neoplastic or malignant cells by inhibiting the growth and/or causing the death of cancer or tumor cells.
[0510] In certain embodiments, the cell-targeting molecule or pharmaceutical composition of the present invention is used to treat a B-cell-, plasma cell- or antibody-mediated disease or disorder, such as for example leukemia, lymphoma, myeloma, Human Immunodeficiency Virus-related diseases, amyloidosis, hemolytic uremic syndrome, polyarteritis, septic shock, Crohn's Disease, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, ulcerative colitis, psoriasis, asthma, Sjgren's syndrome, graft-versus-host disease, graft rejection, diabetes, vasculitis, scleroderma, and systemic lupus erythematosus.
[0511] In another aspect, certain embodiments of the cell-targeting molecules and pharmaceutical compositions of the present invention are antimicrobial agentsmeaning they are capable of treating and/or preventing the acquisition, development, or consequences of microbiological pathogenic infections, such as caused by viruses, bacteria, fungi, prions, or protozoans.
[0512] It is within the scope of the present invention to provide a prophylaxis or treatment for diseases or conditions mediated by T-cells or B-cells by administering the cell-targeting molecule the present invention, or a pharmaceutical composition thereof, to a patient for the purpose of killing T-cells or B-cells in the patient. This usage is compatible with preparing or conditioning a patient for bone marrow transplantation, stem cell transplantation, tissue transplantation, or organ transplantation, regardless of the source of the transplanted material, e.g. human or non-human sources.
[0513] It is within the scope of the present invention to provide a bone marrow recipient for prophylaxis or treatment of host-versus-graft disease via the targeted cell-killing of host T-cells using a cytotoxic cell-targeting molecule or pharmaceutical composition of the present invention.
[0514] Certain embodiments of the cell-targeting molecules and pharmaceutical compositions of the present invention can be utilized in a method of treating cancer comprising administering to a patient, in need thereof, a therapeutically effective amount of a cell-targeting molecule and/or pharmaceutical composition of the present invention. In certain embodiments of the methods of the present invention, the cancer being treated is selected from the group consisting of: bone cancer (such as multiple myeloma or Ewing's sarcoma), breast cancer, central/peripheral nervous system cancer (such as brain cancer, neurofibromatosis, or glioblastoma), gastrointestinal cancer (such as stomach cancer or colorectal cancer), germ cell cancer (such as ovarian cancers and testicular cancers, glandular cancer (such as pancreatic cancer, parathyroid cancer, pheochromocytoma, salivary gland cancer, or thyroid cancer), head-neck cancer (such as nasopharyngeal cancer, oral cancer, or pharyngeal cancer), hematological cancers (such as leukemia, lymphoma, or myeloma), kidney-urinary tract cancer (such as renal cancer and bladder cancer), liver cancer, lung/pleura cancer (such as mesothelioma, small cell lung carcinoma, or non-small cell lung carcinoma), prostate cancer, sarcoma (such as angiosarcoma, fibrosarcoma, Kaposi's sarcoma, or synovial sarcoma), skin cancer (such as basal cell carcinoma, squamous cell carcinoma, or melanoma), and uterine cancer.
[0515] Certain embodiments of the cell-targeting molecules and pharmaceutical compositions of the present invention can be utilized in a method of treating an immune disorder comprising administering to a patient, in need thereof, a therapeutically effective amount of the cell-targeting molecules and/or pharmaceutical composition of the present invention. In certain embodiments of the methods of the present invention, the immune disorder is related to an inflammation associated with a disease selected from the group consisting of: amyloidosis, ankylosing spondylitis, asthma, Crohn's disease, diabetes, graft rejection, graft-vs.-host disease, Hashimoto's thyroiditis, hemolytic uremic syndrome, HIV-related diseases, lupus erythematosus, multiple sclerosis, polyarteritis, psoriasis, psoriatic arthritis, rheumatoid arthritis, scleroderma, septic shock, Sjgren's syndrome, ulcerative colitis, and vasculitis.
[0516] Among certain embodiments of the present invention is using the Shiga toxin effector polypeptide or cell-targeting molecule of the present invention as a component of a pharmaceutical composition or medicament for the treatment or prevention of a cancer, tumor, other growth abnormality, immune disorder, and/or microbial infection. For example, immune disorders presenting on the skin of a patient may be treated with such a medicament in efforts to reduce inflammation. In another example, skin tumors may be treated with such a medicament in efforts to reduce tumor size or eliminate the tumor completely.
[0517] Certain cytotoxic cell-targeting molecules of the present invention, and compositions thereof, may be used in molecular neurosurgery applications such as immunolesioning and neuronal tracing. For example, the targeting domain may be selected or derived from various ligands, such as neurotransmitters and neuropeptides, which target specific neuronal cell types by binding neuronal surface receptors, such as a neuronal circuit specific G-protein coupled receptor. Similarly, the targeting domain may be selected from or derived from antibodies that bind neuronal surface receptors. Because certain Shiga toxin effector polypeptides robustly direct their own retrograde axonal transport, certain cell-targeting molecules of the present invention may be used to kill a neuron(s) which expresses the extracellular target at a site of cytotoxic protein injection distant from the cell body, These targeted cytotoxic molecules of the invention that specifically target neuronal cell types have uses in neuroscience research, such as for elucidating mechanisms of sensations, and creating model systems of neurodegenerative diseases, such as Parkinson's and Alzheimer's.
[0518] Among certain embodiment of the present invention is a method of using a Shiga toxin effector polypeptide, cell-targeting molecule, pharmaceutical composition, and/or diagnostic composition of the present invention to label or detect the interiors of neoplastic cells and/or immune cell types. This method may be based on the ability of certain cell-targeting molecules of the present invention to enter specific cell types and route within cells via retrograde intracellular transport, to the interior compartments of specific cell types are labeled for detection. This can be performed on cells in situ within a patient or on cells and tissues removed from an organism, e.g. biopsy material.
[0519] Among certain embodiment of the present invention is a method of using a Shiga toxin effector polypeptide, cell-targeting molecule, pharmaceutical composition, and/or diagnostic composition of the present invention to detect the presence of a cell type for the purpose of information gathering regarding diseases, conditions and/or disorders. The method comprises contacting a cell with a diagnostically sufficient amount of a cell-targeting molecule of the present invention in order to detect the molecule by an assay or diagnostic technique. The phrase diagnostically sufficient amount refers to an amount that provides adequate detection and accurate measurement for information gathering purposes by the particular assay or diagnostic technique utilized. Generally, the diagnostically sufficient amount for whole organism in vivo diagnostic use will be a non-cumulative dose of between 0.001 to 10 milligrams of the detection-promoting agent linked cell-targeting molecule of the invention per kg of subject per subject. Typically, the amount of Shiga toxin effector polypeptide or cell-targeting molecule of the invention used in these information gathering methods will be as low as possible provided that it is still a diagnostically sufficient amount. For example, for in vivo detection in an organism, the amount of Shiga toxin effector polypeptide, cell-targeting molecule, or pharmaceutical composition of the invention administered to a subject will be as low as feasibly possible.
[0520] The cell-type specific targeting of cell-targeting molecules of the present invention combined with detection-promoting agents provides a way to detect and image cells physically coupled with an extracellular target biomolecule of a binding region of the molecule of the invention. Imaging of cells using the cell-targeting molecules of the present invention may be performed in vitro or in vivo by any suitable technique known in the art. Diagnostic information may be collected using various methods known in the art, including whole body imaging of an organism or using ex vivo samples taken from an organism. The term sample used herein refers to any number of things, but not limited to, fluids such as blood, urine, serum, lymph, saliva, anal secretions, vaginal secretions, and semen, and tissues obtained by biopsy procedures. For example, various detection-promoting agents may be utilized for non-invasive in vivo tumor imaging by techniques such as magnetic resonance imaging (MRI), optical methods (such as direct, fluorescent, and bioluminescent imaging), positron emission tomography (PET), single-photon emission computed tomography (SPECT), ultrasound, x-ray computed tomography, and combinations of the aforementioned (see, Kaur S et al., Cancer Lett 315: 97-111 (2012), for review).
[0521] Among certain embodiment of the present invention is a method of using a Shiga toxin effector polypeptide, cell-targeting molecule, or pharmaceutical composition of the present invention in a diagnostic composition to label or detect the interiors of a hematologic cell, cancer cell, tumor cell, infected cell, and/or immune cell (see e.g., Koyama Y et al., Clin Cancer Res 13: 2936-45 (2007); Ogawa M et al., Cancer Res 69: 1268-72 (2009); Yang L et al., Small 5: 235-43 (2009)). Based on the ability of certain cell-targeting molecules of the invention to enter specific cell types and route within cells via retrograde intracellular transport, the interior compartments of specific cell types are labeled for detection. This can be performed on cells in situ within a patient or on cells and tissues removed from an organism, e.g. biopsy material.
[0522] Diagnostic compositions of the present invention may be used to characterize a disease, disorder, or condition as potentially treatable by a related pharmaceutical composition of the present invention. In certain compositions of matter of the present invention may be used to determine whether a patient belongs to a group that responds to a therapeutic strategy which makes use of a compound, composition or related method of the present invention as described herein or is well suited for using a delivery device of the invention.
[0523] Diagnostic compositions of the present invention may be used after a disease, e.g. a cancer, is detected in order to better characterize it, such as to monitor distant metastases, heterogeneity, and stage of cancer progression. The phenotypic assessment of disease disorder or infection can help prognostic and prediction during therapeutic decision making. In disease reoccurrence, certain methods of the invention may be used to determine if local or systemic problem.
[0524] Diagnostic compositions of the present invention may be used to assess responses to therapies regardless of the type of the type of therapy, e.g. small molecule drug, biological drug, or cell-based therapy. For example, certain embodiments of the diagnostics of the invention may be used to measure changes in tumor size, changes in antigen positive cell populations including number and distribution, or monitoring a different marker than the antigen targeted by a therapy already being administered to a patient (see Smith-Jones P et al., Nat. Biotechnol 22: 701-6 (2004); Evans M et al., Proc. Natl. Acad. Sci. USA 108: 9578-82 (2011)).
[0525] In certain embodiments of the method used to detect the presence of a cell type may be used to gather information regarding diseases, disorders, and conditions, such as, for example bone cancer (such as multiple myeloma or Ewing's sarcoma), breast cancer, central/peripheral nervous system cancer (such as brain cancer, neurofibromatosis, or glioblastoma), gastrointestinal cancer (such as stomach cancer or colorectal cancer), germ cell cancer (such as ovarian cancers and testicular cancers, glandular cancer (such as pancreatic cancer, parathyroid cancer, pheochromocytoma, salivary gland cancer, or thyroid cancer), head-neck cancer (such as nasopharyngeal cancer, oral cancer, or pharyngeal cancer), hematological cancers (such as leukemia, lymphoma, or myeloma), kidney-urinary tract cancer (such as renal cancer and bladder cancer), liver cancer, lung/pleura cancer (such as mesothelioma, small cell lung carcinoma, or non-small cell lung carcinoma), prostate cancer, sarcoma (such as angiosarcoma, fibrosarcoma, Kaposi's sarcoma, or synovial sarcoma), skin cancer (such as basal cell carcinoma, squamous cell carcinoma, or melanoma), uterine cancer, AIDS, amyloidosis, ankylosing spondylitis, asthma, autism, cardiogenesis, Crohn's disease, diabetes, erythematosus, gastritis, graft rejection, graft-versus-host disease, Grave's disease, Hashimoto's thyroiditis, hemolytic uremic syndrome, HIV-related diseases, lupus erythematosus, lymphoproliferative disorders (including post-transplant lymphoproliferative disorders), multiple sclerosis, myasthenia gravis, neuroinflammation, polyarteritis, psoriasis, psoriatic arthritis, rheumatoid arthritis, scleroderma, septic shock, Sjgren's syndrome, systemic lupus erythematosus, ulcerative colitis, vasculitis, cell proliferation, inflammation, leukocyte activation, leukocyte adhesion, leukocyte chemotaxis, leukocyte maturation, leukocyte migration, neuronal differentiation, acute lymphoblastic leukemia (ALL), T acute lymphocytic leukemia/lymphoma (ALL), acute myelogenous leukemia, acute myeloid leukemia (AML), B-cell chronic lymphocytic leukemia (B-CLL), B-cell prolymphocytic lymphoma, Burkitt's lymphoma (BL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CIVIL-BP), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia (HCL), Hodgkin's Lymphoma (HL), intravascular large B-cell lymphoma, lymphomatoid granulomatosis, lymphoplasmacytic lymphoma, MALT lymphoma, mantle cell lymphoma, multiple myeloma (MM), natural killer cell leukemia, nodal marginal B-cell lymphoma, Non-Hodgkin's lymphoma (NHL), plasma cell leukemia, plasmacytoma, primary effusion lymphoma, pro-lymphocytic leukemia, promyelocytic leukemia, small lymphocytic lymphoma, splenic marginal zone lymphoma, T-cell lymphoma (TCL), heavy chain disease, monoclonal gammopathy, monoclonal immunoglobulin deposition disease, myelodusplastic syndromes (MDS), smoldering multiple myeloma, and Waldenstrom macroglobulinemia.
[0526] The present invention is further illustrated by the following non-limiting examples of 1) Shiga toxin effector polypeptides of the present invention, 2) cell-targeting molecules of the present invention, and 3) cytotoxic, cell-targeting molecules of the present invention comprising the aforementioned polypeptides and capable of specifically targeting certain cell types.
EXAMPLES
[0527] The following examples demonstrate certain embodiments of the present invention. However, it is to be understood that these examples are for illustration purposes only and do not intend, nor should any be construed, to be wholly definitive as to conditions and scope of this invention. The experiments in the following examples were carried out using standard techniques, which are well known and routine to those of skill in the art, except where otherwise described in detail.
[0528] Site-specific conjugation strategies based on unique amino acid residues can help control homogeneity of the product(s) of a conjugation reaction, such as, e.g., strategies using engineered cysteine residues, unnatural amino acid residues, and/or enzymatic conjugation. By using unique conjugation sites, heterogeneity may be minimized and the resulting compositions become more consistent from batch to batch and have predictable properties.
[0529] These examples describe the creation and testing of various molecules, each comprising an amino acid residue for site-specific conjugation to various molecules, such as, e.g., a unique, free cysteine or lysine residue in a Shiga toxin effector polypeptide or Shiga toxin effector polypeptide scaffold, whether the unique, free cysteine or lysine residue is positioned ectopically or at a naturally occurring position). In certain examples, the Shiga toxin effector polypeptides are de-immunized as described in WO 2015/113007. In certain examples, the Shiga toxin effector polypeptides comprise an embedded, heterologous, CD8+ T-cell epitope as described in WO 2015/113005. In certain examples, the Shiga toxin effector polypeptides comprise a disrupted furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment derived region that confers reduced protease cleavage sensitivity to the Shiga toxin effector polypeptide as a component of certain molecules as described in WO 2015/191764. In certain examples, the Shiga toxin effector polypeptides are de-immunized, comprise an embedded, heterologous, CD8+ T-cell epitope, and comprise a disrupted furin-cleavage motif at the carboxy-terminus of a Shiga toxin A1 fragment as described in WO 2016/196344.
[0530] Exemplary, Shiga toxin effector polypeptides were tested for the retention of Shiga toxin effector function(s) as components of exemplary, cell-targeting molecules of the present invention. The retention of potent cytotoxicity by a Shiga toxin effector polypeptide component of a cell-targeting molecules implicates the retention by that Shiga toxin effector polypeptide of a certain, minimum level of retrograde intracellular routing to the cytosol. The presence of free cysteine residues in exemplary cell-targeting molecules was demonstrated by the formation of intermolecular disulfide bonds linking cell-targeting protein monomers.
[0531] The presence of one or more ectopic, free cysteine residues and/or a unique lysine residue in a Shiga toxin A Subunit effector scaffold provides an amenable attachment point for linking various molecules to the Shiga toxin effector polypeptide scaffold at specific residues in a controlled fashion. Such linked molecules may be (1) cell-targeting agents; (2) cell-targeting, proteinaceous molecules; (3) cargos designed for intracellular delivery, including for controlled liberation; and/or (4) agents having extracellular function(s), such as, e.g., biologically inert moieties which prolong half-life in a vertebrate and/or mask immunogenic portions of the scaffold. The conjugation of cell-targeting molecules comprising Shiga toxin A Subunit effectors to a cargo (such as a cytotoxic payload) allows for the targeted delivery of the cargo or payload to an internal location of target cells. Examples of cargos include DNA, RNA, nucleic acid complexes, enzymes, proteins, peptides, fluorescent proteins or peptides, and cytotoxic small molecules. Certain examples show exemplary, cell-targeting molecules comprising a Shiga toxin effector polypeptide having an ectopic, cysteine residue conjugated using standard techniques to a cargo, such as, e.g., a peptide, nucleic acid, protein, protein-nucleic acid complex, cytotoxic agent, antibiotic, and/or detection-promoting agent.
Example 1. Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention Comprising a Cysteine Residue(s) for Site-Specific Conjugation
[0532] In this Example, exemplary Shiga toxin A Subunit effector polypeptides of the present invention (SLT-1A-Cys(p)-variant), where Cys(p) represents a cysteine residue engineered at a unique position), each comprising one ectopic cysteine residue, were created and tested as components of exemplary cell-targeting molecules of the present invention.
A. Constructing Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention by Adding Free Cysteine Residues Via Amino Acid Residue Substitution
[0533] This section describes the creation of various scaffolds comprising Shiga toxin effector polypeptides, each comprising one ectopic, cysteine residue. The ectopic, cysteine residues were engineered into Shiga toxin effector polypeptides as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. Exemplary, cell-targeting molecules of the present invention (e.g. SLT-1A-Cys(p)-variant::scFv(n)) were created using these Shiga toxin effector polypeptides.
[0534] In this Example, cell-targeting molecules, each comprising a Shiga toxin A Subunit effector polypeptide having one ectopic, cysteine residue, were created and tested. These cell-targeting molecules each comprised a cell-targeting, immunoglobulin-type, binding region comprising a polypeptide capable of binding to an extracellular target biomolecule with high-affinity.
[0535] The parental, Shiga toxin effector polypeptide of this Example is the A1 fragment of Shiga-like toxin (SLT-1A1) comprising the substitution C242S to remove the only endogenous, cysteine residue (SEQ ID NO:4). The mutation of the cysteine at position 242 to a serine (C242S) had no apparent effect on catalytic activity of the A1 fragment. Using standard techniques, the parental, Shiga toxin effector polypeptide is used to make various Shiga toxin effector polypeptides, each having one ectopic cysteine residue and no other cysteine residues (see e.g. Table 1). Cell-targeting molecules (see e.g. SEQ ID NOs: 773-783) comprising Shiga toxin effector polypeptides (see e.g. SEQ ID NOs: 5-84) having exactly one of the ectopic cysteine residues described in Table 1 were made using standard techniques and tested in one or more of the experiments described below.
TABLE-US-00003 TABLE 1 Ectopic, Cysteine Residues Engineered into Shiga Toxin Effector Polypeptides Exemplary, Shiga toxin Cysteine Sequence Variants effector polypeptides Substitution (SLT-1A-Cys(p)-variant)) SLT-1A-Cys1 K1C SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, and 115 SLT-1A-Cys2 S8C SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, and 116 SLT-1A-Cys3 S16C SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, and 117 SLT-1A-Cys4 S22C SEQ ID NOs: 8, 18, 28, 38, 48, 58, 68, 78, 88, 98, 108, and 118 SLT-1A-Cys5 S33C SEQ ID NOs: 9, 19, 29, 39, 49, 59, 69, 79, 89, 99, 109, 119, and 1101 SLT-1A-Cys6 S43C SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, and 120 SLT-1A-Cys7 S45C SEQ ID NOs: 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, and 1102 SLT-1A-Cys8 S146C SEQ ID NOs: 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, and 1103 SLT-1A-Cys9 S186C SEQ ID NOs: 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 123, and 1104 SLT-1A-Cys10 V54C SEQ ID NOs: 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, and 124
B. Testing Shiga Toxin A Subunit Effector Polypeptides for Retention of Shiga Toxin Functions
[0536] Shiga toxin effector polypeptides and molecules comprising the same can be tested for Shiga toxin functions using techniques known to the skilled worker (see e.g. WO 2014/164680, WO 2014/164693, WO 2015/113005; WO 2015/113007; WO 2015/138435, WO 2015/138452, WO 2015/191764, US20160177284; WO 2016/126950). Exemplary, cell-targeting molecules were tested for Shiga toxin A Subunit functions. The Shiga toxin A Subunit functions analyzed were: catalytic activity, inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol.
1. Testing Shiga Toxin Catalytic Activity and Ability to Inhibit Ribosome Function
[0537] The catalytic activities of Shiga toxin effector polypeptide components of exemplary, cell-targeting molecules were tested using a ribosome inhibition assay. Catalytic activities were tested for exemplary, cell-targeting molecules comprising exemplary, Shiga toxin effector polypeptides of the present invention (SEQ ID NOs: 9, 11, and 14).
[0538] The ribosome inactivation capabilities of exemplary, cell-targeting molecules of this Example were determined using a cell-free, in vitro protein translation assay based on the TNT Quick Coupled Transcription/Translation Kit (L1170, Promega Corp., Madison, WI, U.S.). The ribosome activity reaction was prepared according to manufacturer's instructions. A series of 10-fold dilutions of the cell-targeting molecule to be tested was prepared in an appropriate buffer and a series of identical, TNT reaction mixtures were created for each dilution. Each sample in the dilution series was combined with each of the TNT reaction mixtures along with Luciferase T7 Control DNA (L4821, Promega Corp., Madison, WI, U.S.). The test samples were incubated for 1.5 hours at 30 degrees Celsius ( C.). After the incubation, Luciferase Assay Reagent (E1483, Promega Corp., Madison, WI, U.S.) was added to all test samples and the amount of luciferase protein present was measured by luminescence according to manufacturer's instructions. A positive control was tested: the wild-type, Shiga-like toxin A1 fragment (SLT-1A1-WT) (SEQ ID NO:830). This cell-free, in vitro, protein translation assay was used to determine the ribosome inactivation capabilities of SLT-1A-Cys5::scFv1 (SEQ ID NO:781), SLT-1A-Cys7::scFv1 (SEQ ID NO:782), and SLT-1A-Cys10::scFv1 (SEQ ID NO:783).
[0539] The level of translational inhibition was determined by non-linear regression analysis of log-transformed concentrations of total protein versus relative luminescence units (RLU). Using Prism software (GraphPad Prism, San Diego, CA, U.S.), the half maximal inhibitory concentration (IC.sub.50) value was calculated for each sample using the Prism software function of log(inhibitor) vs. response (three parameters) [Y=Bottom+((TopBottom)/(1+10{circumflex over ()}(XLog IC.sub.50)))] under the heading dose-response-inhibition. The IC.sub.50 values for each cell-targeting molecule from one or more experiments was calculated and is shown in Table 2 in picomolar (pM). In this assay, measurements of the inhibition of protein synthesis represent the ribosome inactivation activity of the sample molecule, which is one metric of the catalytic activity of a Shiga toxin effector polypeptide. Any molecule which exhibits an IC.sub.50 within 10-fold of a reference molecule is considered herein to exhibit ribosome inhibition activity comparable to the reference molecule. As reported in the Examples, a molecule exhibiting an IC.sub.50 less than or within 10 percent of an IC.sub.50 exhibited by a reference molecule is considered to exhibit ribosome inhibition activity equivalent to that reference molecule.
TABLE-US-00004 TABLE 2 Ribosome Inhibition Activities of Exemplary, Cell-Targeting Molecules Cysteine Ribosome Inhibition Molecule Tested position IC.sub.50 (pM) SLT-1A-Cys5::scFv1 C33 190 (SEQ ID NO: 781) SLT-1A-Cys7::scFv1 C45 109 (SEQ ID NO: 782) SLT-1A-Cys10::scFv1 C54 78 (SEQ ID NO: 783) SLT-1A1-WT (wild-type) none 185 (SEQ ID NO: 830)
[0540] The ribosome inactivation activities of all the Shiga toxin effector polypeptides SLT-1A-Cys(p) tested in the context of a cell-targeting molecule were equivalent to the catalytic activity of a wild-type, Shiga toxin A1 fragment (Table 2). As shown in Table 2, exemplary cell-targeting molecules comprising or consisting of SEQ ID NO: 781, 782, or 783 exhibited potent ribosome inhibition equivalent to the positive control of an isolated Shiga toxin A Subunit effector polypeptide consisting of a wild-type, Shiga toxin A1 fragment (SEQ ID NO:830). The results of this assay showed that multiple, exemplary, Shiga toxin effector polypeptides of the present invention (SEQ ID NOs: 9, 11, and 14) exhibited catalytic activity in the context of a cell-targeting molecule equivalent to the activity of a wild-type, Shiga toxin effector polypeptide that comprised a single, endogenous cysteine residue at position 242 (Table 2). In other experiments, it was demonstrated that cell-targeting molecules comprising mutated Shiga toxin effector polypeptides having no cysteine residues or only a single free cysteine residue, the endogenous cysteine residue at position 242 (e.g. a wild-type SLT-1A1 (amino acids 1-251 of SEQ ID NO:1)), exhibited catalytic activity comparable to the activity of each other and to the activity of a cell-targeting molecule comprising a wild-type, Shiga toxin effector polypeptide (amino acids 1-251 of SEQ ID NO:1).
2. Testing the Cytotoxic Activities of Exemplary, Cell-Targeting Molecules of the Invention
[0541] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules were tested to assess the functions of their Shiga toxin effector polypeptide components. The cytotoxic activities of exemplary, cell-targeting molecules, which each comprised a Shiga toxin effector polypeptide SLT-1A-Cys(p), were determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker (cell-kill assay). The cytotoxicities of exemplary, cell-targeting molecules were determined using cells expressing, at a cellular surface, significant amounts of the appropriate, extracellular target biomolecule, such as, a target of the binding region scFv1, scFv2, scFv3, and/or scFv4. The cells used in this Example were immortalized, human tumor cells available from the ATCC (Manassas VA, U.S.), National Cancer Institute of the U.S. (Frederick, MD, U.S.), and/or DSZM (Braunschweig, DE). The cells referred to below were HCC-1954, MDA-MB-231, H929, ST486, L1236, HCC827, Daudi, HDLM-2, and U-266, or more simply cell-lines A, B, C, D, E, F, G, H and I, respectively. Certain cell-targeting molecules were tested using cell-kill assays involving both target biomolecule positive and target biomolecule negative cells with respect to the target biomolecule of each cell-targeting molecule's binding region.
[0542] The cytotoxicity assays were performed as follows. Certain, human tumor cell-line cells were plated (typically at 210.sup.3 cells per well for adherent cells, plated the day prior to cell-targeting molecule addition, or 7.510.sup.3 cells per well for suspension cells, plated the same day as cell-targeting molecule addition) in 20 L cell culture medium in 384-well plates. A series of 10-fold dilutions of the molecules to be tested was prepared in an appropriate buffer, and 5 L of the dilutions or buffer control were added to the plated cells. Control wells containing only cell culture medium were used for baseline correction. The cell samples were incubated with the cell-targeting molecule or just buffer for 3 or 5 days at 37 C. and in an atmosphere of 5% carbon dioxide (CO.sub.2). The total cell survival or percent viability was determined using a luminescent readout using the CellTiter-Glo Luminescent Cell Viability Assay or RealTime-Glo MT Cell Viability Assay (Promega Corp., Madison, WI, U.S.) according to the manufacturer's instructions.
[0543] The Percent Viability of experimental wells was calculated using the following equation: (Test RLUAverage Media RLU)(Average Cells RLUAverage Media RLU)100. The logarithm of the cell-targeting molecule protein concentration versus Percent Viability was plotted in Prism (GraphPad Prism, San Diego, CA, U.S.) and log (inhibitor) versus response (3 parameter) analysis or and log (inhibitor) versus normalized response analysis were used to determine the half-maximal cytotoxic concentration (CD50) value for the tested molecule. The CD.sub.50 value(s) for each molecule tested were calculated, when possible, and shown in Table 3. When CD.sub.50 values could not be calculated based on the shape of the curve over the concentrations tested, then a maximum CD.sub.50 value was noted as being beyond the maximum tested value, e.g., greater than 100 nanomolar (>100 nM) or 200 nanomolar (>200 nM), for samples which did not kill 50% of the cells at the highest, tested, sample concentration, e.g., 100 or 200 nanomolar (nM). In some experiments, biomolecule target negative cells treated with the maximum concentration of the cell-targeting molecule did not show any change in viability as compared to a buffer only control. As reported in the Examples herein, a molecule exhibiting a CD.sub.50 within 10-fold of a CD.sub.50 exhibited by a reference molecule is considered to exhibit cytotoxic activity comparable to that reference molecule. Cell-targeting molecules that exhibited a CD.sub.50 to a biomolecule target positive cell population within 100-fold to 10-fold of a reference molecule comprising the same binding region and a related Shiga toxin effector polypeptide component lacking any cysteine residue is referred to herein as active but attenuated. The CD.sub.50 values for exemplary, cell-targeting molecules are shown in Table 3 and associated cell-kill assay data is shown in
TABLE-US-00005 TABLE 3 Cytotoxic Activities of Exemplary, Cell-Targeting Molecules SLT-1A Cell-type tested Cell-type tested Cysteine (target positive Cytotoxicity (target positive Cytotoxicity Molecule tested position or negative) CD.sub.50 (nM) or negative) CD.sub.50 (nM) Experiment 1 SLT-1A- C33 Cell Line A 0.67 Cell Line B >200 nM Cys5::scFv1 (positive) (negative) SLT-1A- C45 Cell Line A 0.44 Cell Line B >200 nM Cys7::scFv1 (positive) (negative) SLT-1A- C54 Cell Line A 0.42 Cell Line B >200 nM Cys10::scFv1 (positive) (negative) SLT-1A1-WT C242 Cell Line A 373.00 Cell Line B >200 nM (n/a) (n/a) Experiment 2 SLT-1A-Cys2- C8 Cell Line C 0.0023 Cell Line D 0.0074 D1::scFv2 (positive) (positive) SLT-1A-Cys6- C43 Cell Line C 0.0041 Cell Line D 0.0588 D1::scFv2 (positive) (positive) SLT-1A-Cys7- C45 Cell Line C 0.0012 Cell Line D 0.0063 D1::scFv2 (positive) (positive) SLT-1A-Cys8- C146 Cell Line C 0.0010 Cell Line D 0.0038 D1::scFv2 (positive) (positive) SLT-1A-Cys9- C186 Cell Line C 0.0046 Cell Line D 0.0176 D1::scFv2 (positive) (positive) SLT-1A-D1:: none Cell Line C 0.0026 Cell Line D 0.0073 scFv2 (positive) (positive) SLT-1A-D1 none Cell Line C >100 nM Cell Line D >100 nM (n/a) (n/a) Experiment 3 SLT-1A-Cys2- C8 Cell Line E 54.8 D1::scFv3 (positive) SLT-1A-Cys3- C16 Cell Line E 34.8 D1::scFv3 (positive) SLT-1A-Cys5- C33 Cell Line E 17.4 D1::scFv3 (positive) SLT-1A-D1- C242 Cell Line E 40.3 C242::scFv3 (positive) SLT-1A- none Cell Line E 55.7 D1::scFv3 (positive) * n/a denotes that the presence or absence of a particular target biomolecule is irrelevant because the molecule tested was untargeted
[0544] The exemplary cell-targeting molecules of the present invention represented by SLT-1A-Cys5::scFv1 (SEQ ID NO:781), SLT-1A-Cys7::scFv1 (SEQ ID NO:782), SLT-1A-Cys10::scFv1 (SEQ ID NO:783), SLT-1A-Cys2-D1::scFv2 (SEQ ID NO:773), SLT-1A-Cys2-D1::scFv3 (SEQ ID NO:780), SLT-1A-Cys3-D1::scFv3 (SEQ ID NO:779), SLT-1A-Cys5-D1::scFv3 (SEQ ID NO:778), SLT-1A-Cys6-D1::scFv2 (SEQ ID NO:774), SLT-1A-Cys7-D1::scFv2 (SEQ ID NO:775), SLT-1A-Cys8-D1::scFv2 (SEQ ID NO:776), and SLT-1A-Cys9-D1::scFv2 (SEQ ID NO:777) were each cytotoxic to target positive cells, commonly exhibiting CD.sub.50 values less than 100 nM. The results reported in Table 3 show that cell-targeting molecules comprising any of the Shiga toxin effector polypeptides SLT-1A-Cys5 (SEQ ID NO:9), SLT-1A-Cys7 (SEQ ID NO:11), SLT-1A-Cys10 (SEQ ID NO:14) were cytotoxic to target positive cells at potencies associated with CD.sub.50 values generally less than 100 nM to 5 micromolar (04) depending on the cell-line tested (see Table 3;
[0545]
[0546]
[0547] The specificity of the cytotoxic activity of a given cell-targeting molecule can be determined by comparing cell-kill activities toward cells expressing a significant amount of a target biomolecule of the binding region of the cell-targeted molecule (target positive cells) with cell-kill activities toward cells which do not have any significant amount of any target biomolecule of the binding region of the cell-targeted molecule physically coupled to any cellular surface (target negative cells). This was accomplished by comparing the CD.sub.50 value(s) of a given cell-targeting molecule toward a cell population(s) which were positive for cell surface expression of the target biomolecule of the cell-targeting molecule being analyzed to the CD.sub.50 value(s) of the same molecule toward a cell population(s) which were negative for cell surface expression of any target biomolecule of the molecule being analyzed.
[0548] The results of the cell-kill assays described above and shown in Table 3 and
Example 2. Exemplary, Cell-Targeting Molecules of the Present Invention that Comprise a Free Cysteine Residue Outside the Toxin Effector Region
[0549] In this Example, exemplary cell-targeting molecules of the present invention, each comprising one free cysteine residue outside any toxin effector region, were created and tested.
A. Constructing Exemplary, Cell-Targeting Molecules of the Present Invention that Comprise a Free Cysteine Residue Outside all Shiga Toxin Effector Polypeptide Components
[0550] This section describes the creation of various cell-targeting molecules comprising a free cysteine residue. Using standard techniques, cysteine residues were engineered into cell-targeting proteins as genetically encoded substitutions which did not change the overall number of amino acid residues in the cell-targeting protein. In this Example, each cell-targeting molecule that was tested comprised a cell-targeting, immunoglobulin-type, binding region comprising a polypeptide capable of binding to an extracellular target biomolecule with high-affinity (see Table 4). Protein expression and purification of these cell-targeting molecules was performed using standard techniques known to the skilled worker and/or as previously described (see e.g. WO 2014/164680, WO 2014/164693, WO 2015/138435, WO 2015/138452, WO 2015/191764, and WO 2016/126950), such as using Capto-L (GE Healthcare, Marlborough, MA, U.S.).
TABLE-US-00006 TABLE 4 Free Cysteine Residues Engineered into Cell-Targeting Proteins Cell-Targeting Exemplary, Cell- Molecule Targeting Molecules Component Free Cysteine location with Component linker-Cys1 between Binding Region SEQ ID NOs: 803-806 (SEQ ID NO: 757) and Toxin Effector Region linker-Cys2 between Binding Region (SEQ ID NO: 758) and Toxin Effector Region linker-Cys3 between Binding Region (SEQ ID NO: 759) and Toxin Effector Region linker-Cys4 between Binding Region (SEQ ID NO: 760) and Toxin Effector Region scFv-linker-Cys1 between the scFv's heavy SEQ ID NOs: 807-808 (SEQ ID NO: 769) and light variable regions scFv-linker-Cys2 between the scFv's heavy SEQ ID NOs: 809-811 (SEQ ID NO: 770) and light variable regions scFv-linker-Cys3 between the scFv's heavy (SEQ ID NO:771) and light variable regions scFv-linker-Cys4 between the scFv's heavy (SEQ ID NO: 772) and light variable regions scFv(n)-Cys-C2 penultimate C-terminal SEQ ID NOs: 812-815 residue of Binding Region V.sub.HH(n)-Cys-C2 penultimate C-terminal SEQ ID NOs: 816-817 residue of Binding Region
B. Testing the Cytotoxic Activities of Exemplary, Cell-Targeting Molecules of the Invention
[0551] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of the present invention were tested to assess the functionalities of their Shiga toxin effector polypeptide components. The cytotoxic activities of exemplary, cell-targeting molecules, which each comprised one of the following components: linker-Cys(p), scFv(n)-linker-Cys(p), or scFv(n)-Cys(p) were determined using the cell-kill assay described in Example 1 (above). The cytotoxicities of exemplary, cell-targeting molecules were determined using cells expressing, at a cellular surface, significant amounts of the appropriate, extracellular target biomolecule of scFv2. Using the cytotoxicity assay, the CD.sub.50 value(s) for each molecule tested were calculated (Table 5), and associated cell-kill assay data is shown in
TABLE-US-00007 TABLE 5 Cytotoxic Activities of Exemplary, Cell-Targeting Molecules Free Cell-type tested Cytotoxicity Cell-type tested Cytotoxicity Cysteine (target positive CD.sub.50 (target positive CD.sub.50 Molecule Tested location or negative) (nM) or negative) (nM) SLT-1A-D1:: between Cell Line C 0.0006 Cell Line D 0.0089 linker-Cys1::scFv2 Toxin (positive) (positive) (SEQ ID NO: 803) Effector and Binding Region SLT-1A-D1:: between Cell Line C 0.0038 Cell Line D 0.0273 scFv2-linker-Cys1 variable (positive) (positive) (SEQ ID NO: 807) domains of scFv2 SLT-1A-D1:: in variable Cell Line C 0.0002 Cell Line D 0.002 scFv2-Cys-C2 domain of (positive) (positive) (SEQ ID NO: 812) scFv2 SLT-1A-D1::scFv2 none Cell Line C 0.0026 Cell Line D 0.0073 (SEQ ID NO: 838) (positive) (positive) SLT-1A-D1 none Cell Line C >100 nM Cell Line D >100 nM (SEQ ID NO: 831) (n/a) (n/a)
[0552] Table 5 and
Example 3. Testing Purified, Cell-Targeting Molecules for Intermolecular Disulfide Bonding
[0553] Certain, exemplary, cell-targeting molecules of this Example where produced and purified using routine methods known to the skilled worker and then analyzed for intermolecular associations. The cell-targeting molecules tested in this Example included those which comprise a single free cysteine residue such as #1) SLT-1A-D1-C242::scFv3 (SEQ ID NO:838), #2) SLT-1A-Cys2-D1::scFv2 (SEQ ID NO:773), #3) SLT-1A-Cys2-D1::scFv3 (SEQ ID NO:780), #4) SLT-1A-Cys3-D1::scFv3 (SEQ ID NO:779), #5) SLT-1A-Cys5-D1::scFv3 (SEQ ID NO:778), #6) SLT-1A-Cys6-D1::scFv2 (SEQ ID NO:774), #7) SLT-1A-Cys7-D1::scFv2 (SEQ ID NO:775), #8) SLT-1A-Cys8-D1::scFv2 (SEQ ID NO:776), #9) SLT-1A-Cys9-D1::scFv2 (SEQ ID NO:777), #10) SLT-1A-D1::linker-Cys1::scFv2 (SEQ ID NO:803), #11) SLT-1A-D1::scFv2-linker-Cys1 (SEQ ID NO:807), and #12) SLT-1A-D1::scFv2-Cys-C2 (SEQ ID NO:812) as well as other cell-targeting molecules used as various controls such as SLT-1A-D1::scFv2 (SEQ ID NO:838) and SLT-1A-D1::scFv3 (SEQ ID NO:839).
[0554] Exemplary, cell-targeting molecule preparations were analyzed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (SDS-PAGE). Exemplary, cell-targeting molecules and the reference molecules SLT-1A-D1::scFv(n) were loaded in equal concentrations to replicate, 4-20%, SDS-PAGE gels (Lonza, Basel, CH) and electrophoresed under denaturing conditions or denaturing and reducing conditions. The resulting gels were analyzed by COOMASSIE staining for intermolecular associations indicative of disulfide bond formation (shown in
[0555] In
[0556] In
[0557] In
[0558] The results of this analysis and as shown in
[0559] To further analyze the nature of intermolecular associations formed among purified, exemplary, cell-targeting molecules, two samples were further purified and analyzed. Samples of the exemplary, cell-targeting molecules SLT-1A-Cys2-D1::scFv2 and SLT-1A-Cys7-D1::scFv2 used in the experiments above were run over a size exclusion chromatography (SEC) column (Superdex 200 Increase, GE Healthcare, Marlborough, MA, U.S.) and compared to commercial, molecular-size migration reference standards to estimate sizes (see e.g.
[0560] Molecules of known sizes and migration characteristics (standards) were analyzed in order to calibrate which retention times corresponded to which protein sizes and/or protein sizes were predicted from the amino acid composition and/or SDS-PAGE analyses of the purified protein present in the sample to be analyzed. Chromatographic data collected from commercial size standards were used to create calibration curves to help estimate sizes of molecular species in samples and focus analyses on specific retention time ranges.
[0561] The results of the experiments shown in
[0562] The results of the experiments shown in
Example 4. Constructing Cargo-Linked, Cell-Targeting Molecules Comprising Exemplary, Shiga Toxin A Subunit Effector Polypeptides of the Present Invention
[0563] Exemplary cell-targeting molecules of the present invention (SEQ ID NOs: 789, 791-793, 804, 808, 813, and 1141) were produced, purified and conjugated to a maleimide-activated fluorescent dye using routine methods known to the skilled worker. The maleimide-activated fluorescent dyes used were Alexa Fluor 488 C5 Maleimide and Alexa Fluor 555 C2 Maleimide (Thermo Fisher Scientific, Waltham, MA, U.S.).
[0564] For each sample, 200 microgram (m) of an exemplary cell-targeting molecule was incubated with 2 mM TCEP at pH 7 and room temperature for twenty minutes. A 5-fold molar excess of either the 488 or 555 maleimide-activated dye was added to each sample, and chemical reactions were allowed to proceed at room temperature for one hour. Then for each sample, protein was separated from unbound dye using a purification resin according to the manufacturer's instructions (Antibody Conjugate Purification Kit, Catalog #A33088, Thermo Fisher Scientific, Waltham, MA, U.S.), and the protein concentration was estimated by measuring each sample's absorbance of light of a wavelength of 280 nM.
[0565] The method of conjugation used (i.e. the chemical reaction during the one-hour incubation period) was intended to link the maleimide functional group of the dye to a free cysteine residue of a polypeptide component of the cell-targeting molecule. As a control, a related cell-targeting molecule lacking any free cysteine was subjected to the same chemical reaction and method steps described above.
[0566] In certain embodiments, exemplary cell-targeting molecules of the present invention comprise a mutation known in the art to cause the inactivation of the toxin effector component(s) of the cell-targeting molecule. For example, Shiga toxin effector polypeptides comprising the mutation E167D, wherein 167 refers to the native position of a Shiga toxin A Subunit, is referred to in the Examples herein as IA-SLT-1A, with the IA indicating catalytically inactive or catalytically impaired for noncytotoxic or reduced-cytotoxicity variants as a result of severely reduced catalytic activity consistent with descriptions in the scientific literature and as would be recognized by the skilled worker (see e.g. Hovde C et al., Proc Natl Acad Sci USA 85: 2568-72 (1988); Jackson M et al., J Bacteriol 172: 3346-50 (1990); Gordon V et al., Infect Immun 60: 485-90 (1992); Ohmura M et al., Microb Pathog 15: 169-76 (1993)). Exemplary, cell-targeting molecules were created and tested in Example 5, including cell-targeting molecules comprising a Shiga toxin effector polypeptide(s) comprising the E167D mutation: IA-SLT-1A-D1 (SEQ ID NO:834), IA-SLT-1A-Cys5-D1 variant 2 (SEQ ID NO:1101), IA-SLT-1A-Cys7-D1 (SEQ ID NO:1102), IA-SLT-1A-Cys8-D1 (SEQ ID NO:1103), or IA-SLT-1A-Cys9-D1 (SEQ ID NO:1104)).
Example 5. Testing Cargo-Linked Cell-Targeting Molecules of the Present Invention that Comprise a Cargo Linked to a Site-Specific Free Cysteine Residue
[0567] Cargo-linked cell-targeting molecules were tested for cell-targeting molecule functions (e.g. target binding, cell binding, cellular internalization, intracellular routing efficiency, and cell-killing) using standard techniques known to the skilled worker (see e.g. WO 2014/164680, WO 2014/164693, WO 2015/113005, WO 2015/113007, WO 2015/138435, WO 2015/138452, US 2015/0259428, and WO 2015/191764). Exemplary, cell-targeting molecules were tested for Shiga toxin A Subunit functions of cellular internalization and cytotoxicity, and by inference catalytic activity, inhibition of eukaryotic ribosome function, and self-directing subcellular routing to the cytosol. The potency and specificity of cytotoxic activities of exemplary, cargo-linked cell-targeting molecules of the present invention were tested based on the exemplary cell-targeting molecule comprising or consisting of SEQ ID NO:789. In addition, cellular internalization of cargo by cargo-linked cell-targeting molecules was tested to demonstrate cargo delivery functionality of cell-targeting molecules which each comprised a cargo linked to a specific cysteine residue.
A. Testing Cargo-Linked Cell-Targeting Molecules for Retention of Cell-Targeting
[0568] Exemplary, cargo-linked cell-targeting molecules were tested for cell binding, and by inference target biomolecule binding function, using routine techniques known to the skilled worker. The cell-binding ability of each dye-linked cell-targeting molecule was measured by standard flow cytometry assays known to the skilled worker (e.g. fluorescence-activated cell sorting (FACS)) using immortalized human cell lines that either expressed at a cellular surface the target of the cell-targeting molecule (cell lines C and G) or, as a negative control, cells that did not express the target of the cell-targeting molecule being tested on the cell surface (Cell line H).
[0569] For each sample and cell-type tested, 300,000 cells were incubated on ice for one hour in media containing 200 nM of each dye-linked cell-targeting molecule sample. Then the cells were washed, re-suspended in PBS, and analyzed using a BD Accuri C6 flow cytometer according to the manufacturer's instructions (BD Biosciences, San Jose, CA, U.S.). As assay controls for each specific target, each cell line was tested in the same assay using a monoclonal antibody specific to the same target conjugated to fluorescein isothiocyanate (FITC) as a positive control and an isotype negative monoclonal antibody specific to the same target conjugated to FITC as a negative control. Thus, 300,000 cells from each cell line tested were incubated on ice for one hour in media containing one g of either a positive control antibody or a negative control antibody and analyzed with the same method and parameters. The FACS analysis of each sample involved the sorting of living cells by gating based on forward versus side scatter. The FACS gate for positive was based on the isotype negative control samples (encompassing the negative population) and confirmed with a positive control monoclonal antibody (e.g. anti-target2 mAb-FITC).
[0570] The exemplary cell-targeting molecules tested in this assay included: IA-SLT-1A-Cys5-D1::scFv2 (SEQ ID NO:1141), SLT-1A-Cys5-D1::scFv2 (SEQ ID NO:789), IA-SLT-1A-Cys7-D1::scFv2 (SEQ ID NO:793), IA-SLT-1A-Cys8-D1::scFv2 (SEQ ID NO:791), IA-SLT-1A-Cys9-D1::scFv2 (SEQ ID NO:792), IA-SLT-1A-DI::linker-Cys1::scFv2 (SEQ ID NO:804), IA-SLT-1A-DI::scFv2-linker-Cys1 (SEQ ID NO:808), and IA-SLT-1A-DI::scFv2-Cys1 (SEQ ID NO:813). The results of the FACS cell-binding assay measurements for cells incubated with 200 nM of cell-targeting molecule or control antibody are listed in Table 6 and some representative FACS histogram overlay data is shown in
TABLE-US-00008 TABLE 6 Exemplary, Cargo-Linked Cell-Targeting Molecules Exhibited Binding to Target Positive Cells at Cell-Targeting Molecule Concentrations of 200 nM target target positive target positive negative Cell Line C Cell Line G Cell Line H Molecule Tested by target target target linking to dye and Cysteine positive positive positive assaying cell binding position percentage iMFI percentage iMFI percentage IA-SLT-1A-D1::scFv2 none 5% 1,130 6% 361 7% (SEQ ID NO: 843) IA-SLT-1A-Cys5- 33 99% 52,462 100% 83,443 9% D1::scFv2 (SEQ ID NO: 1141) SLT-1A-Cys5- 33 99% 52,466 100% 92,173 7% D1::scFv2 (SEQ ID NO: 789) IA-SLT-1A-Cys7- 45 98% 39,793 100% 81,658 7% D1::scFv2 (SEQ ID NO: 793) IA-SLT-1A-Cys8- 146 89% 26,388 100% 103,075 7% D1::scFv2 (SEQ ID NO: 791) IA-SLT-1A-Cys9- 186 97% 33,677 100% 60,459 6% D1::scFv2 (SEQ ID NO: 792) IA-SLT-1A-DI::linker- linker 97% 37,627 100% 101,105 6% Cys1::scFv2 (SEQ ID NO: 804) IA-SLT-1A-DI::scFv2- linker 99% 64,287 100% 131,920 6% linker-Cys1 (SEQ ID NO: 808) IA-SLT-1A-DI::scFv2- scFv-Cys1 90% 18,634 100% 35,528 5% Cys1 (SEQ ID NO: 813) anti-target 2 mAb N/A 98% 48,505 100% 156,783 2%
[0571] The results in Table 6 demonstrated that exemplary, cargo-linked cell-targeting molecules bound to target positive cells and labeled them with their dye-cargos. However, the control protein IA-SLT-1A-D1::scFv2 (SEQ ID NO:843), which lacks any free cysteine residue, was not capable of labeling cells after being subjected to the same dye-conjugation procedure (Table 6). The exemplary, cargo-linked cell-targeting molecules did not label appreciable numbers of target negative cells (Table 6).
[0572] In addition, the cell-binding results summarized in Table 6 indicate that some cargo and cell-targeting molecule pairs were not as successful as others. For example, the samples which resulted in lower MFI values and iMFI for target positive cell lines suggest those samples had less efficient maleimide-cysteine conjugation reactions and/or lower effective cell binding properties than other samples tested.
[0573] The results of the FACS cell-binding assay measurements for cells incubated with 200 nM of cell-targeting molecule or antibody are shown in histogram overlays in
[0574] These FACS results show that exemplary, cell-targeting molecules SLT-1A-Cys5-D1::scFv2 (SEQ ID NO:789) and IA-SLT-1A-Cys5-D1::scFv2 (SEQ ID NO:1141) bind target positive cells after conjugation to an Alexa Fluor dye cargo (Table 6;
B. Testing Cargo-Linked Cell-Targeting Molecules for Cell Internalization of their Cargos
[0575] Exemplary, cargo-linked cell-targeting molecules were tested for cellular internalization using routine microscopy-based fluorescent techniques known to the skilled worker (see e.g. WO 2014/164680; WO 2015/138452; US20150259428). The behavior and localization of a fluorescent molecule administered to a cell(s) may be observed using fluorescent microscopy. To examine cellular internalization of the exemplary, cell-targeting molecules of the present invention, a fluorescence microscopy-based assay was used to observe the localization of cargo-linked cell-targeting molecules after administration to cells. The molecules tested in this section were an active or inactive version of a scFv2 targeted molecule (SLT-1A-Cys5-D1::scFv2 (SEQ ID NO:789) and IA-SLT-1A-Cys5-D1::scFv2 (SEQ ID NO:1141)), which comprised the Shiga toxin effector polypeptide of either SLT-1A-Cys5-D1 (SEQ ID NO:29) or IA-SLT-1A-Cys5-D1 variant 2 (SEQ ID NO:1101) conjugated to an Alexa Fluor dye, such as, ALEXA-488 or ALEXA-555.
[0576] In this cell-internalization assay, approximately 500,000 cells per sample were incubated at 37 C. for 1 hour in media containing 200 nM of an exemplary, dye-linked, cell-targeting molecule, a cocktail of protease inhibitors, and human BD Fc Block. The cell samples were then washed and fixed with BD Cytofix/Cytoperm (BD Biosciences, San Jose, CA, U.S.). Then the cell samples were re-suspended in PBS and spread on poly-L-lysine coated glass slides. For each slide, a coverslip was mounted using a 4,6-diamidino-2-phenylindole (DAPI)-containing VECTASHIELD solution (catalog #NC9524612, Fisher Scientific, Waltham, MA, U.S.), and the slides were viewed using fluorescent microscopy.
[0577]
[0578] Both ALEXA-488 and ALEXA-555 conjugated cell-targeting molecules produced similar qualitative results, though the ALEXA-555 signal appeared more intense than the ALEXA-488 signal. The results from this cell internalization assay, some of which are shown in
C. Testing Cell-Targeting Molecules for Conjugation Using a Size Shift
[0579] Cargo-linked cell-targeting molecule samples were analyzed by SDS-PAGE in attempt to observe conjugates having a greater mass than unconjugated molecules.
[0580]
D. Testing Cargo-Linked Cell-Targeting Molecules for Retention of Cytotoxicity
[0581] Exemplary, cargo-linked cell-targeting molecules were tested for the Shiga toxin A Subunit function of cell-targeted cytotoxicity, and by inference catalytic activity, inhibition of eukaryotic ribosome function, and self-directing subcellular routing to the cytosol. The cytotoxicity assay was performed as described in Example 1 and Example 2. Table 7 and
TABLE-US-00009 TABLE 7 Exemplary, Cargo-Linked Cell-Targeting Molecules Exhibited Potent and Specific Cytotoxicity to Target Positive Cells SLT-1A target positive cells target negative Cysteine Cell Line C Cell Line G Cell Line H Molecule Tested position Cargo IC.sub.50 (ng/mL) IC.sub.50 (ng/mL) IC.sub.50 (ng/mL) SLT-1A-D1:: none none 1.11 22 >10,000 scFv2 (SEQ ID NO: 838) SLT-1A-Cys5-D1:: 33 none 0.67 12 >10,000 scFv2 (SEQ ID NO: 789) SLT-1A-Cys5-D1:: 33 ALEXA-488 1.32 48 >10,000 scFv2_ALEXA-488 SLT-1A-Cys5-D1:: 33 ALEXA-555 1.93 71 >10,000 scFv2_ALEXA-555
[0582] The results of the cytotoxicity assay shown in
[0583] The results of the experiments in Examples 4-5 demonstrated that a cargo molecule can be conjugated to a free cysteine residue of a cell-targeting molecule of the present invention such that a useful level of (1) the cell-targeting, target binding function of the binding region component is present, (2) cargo molecule delivery to the interior of a target positive cell, and/or (3) the cytotoxic function of the Shiga toxin A Subunit effector polypeptide component is present.
Example 6. Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention Comprising a Unique Lysine Residue for Site-Specific Conjugation
[0584] This example describes the creation of various scaffolds comprising Shiga toxin effector polypeptides, each comprising one, unique, lysine residue (SLT-1A-Lys(p)-variant), where Lys(p) represents a lysine residue at a unique position) were created and tested as components of exemplary cell-targeting molecules of the present invention. All other lysine residues were removed from the Shiga toxin effector polypeptides as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. Exemplary, cell-targeting molecules of the present invention (e.g. SLT-1A-Lys(p)::scFv(n)) were created using these Shiga toxin effector polypeptides.
[0585] The parental, Shiga toxin effector polypeptide of this Example is the A Subunit of Shiga-like toxin 1 (SEQ ID NO:1). Using standard techniques, the parental, Shiga toxin effector polypeptide are used to make various Shiga toxin effector polypeptides, each having one unique lysine residue (see e.g. Table 8). Cell-targeting molecules (see e.g. SEQ ID NOs: 818-823) comprising Shiga toxin effector polypeptides (see e.g. SEQ ID NOs: 197, 198, 200-202, and 205) having exactly one of the unique lysine residues described in Table 8 were made using standard techniques and tested for retention of Shiga toxin A Subunit functions as described in Example 1.
TABLE-US-00010 TABLE 8 Shiga Toxin Effector Polypeptides Engineered to have Unique Lysine Residues Unique Exemplary, Shiga Toxin Lysine Sequence Variants Effector Polypeptide Residue (Shiga Toxin A Subunit-Lys(p)-variant) SLT-1A-Lys1 K1 SEQ ID NOs: 125-128, 134-137, 143-146, 152-155, 161-164, 170-173, 179-182, 188-191, 197-200, 206-209, 215-218, and 224-227 SLT-1A-Lys2 K11 SEQ ID NOs: 129-133, 138-142, 147-151, 156-160, 165-169, 174-178, 183-187, 192-196, 201-205, 210-214, 219-223, and 228-232
[0586] Exemplary, cell-targeting molecules of the present invention were tested for retention of Shiga toxin A Subunit functions as described in Example 1 after removal of an endogenous lysine residue from their Shiga toxin effector polypeptide components. The Shiga toxin A Subunit functions analyzed were: inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol.
[0587] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of this Example, which each comprised a Shiga toxin effector polypeptide SLT-1A-Lys(p), were determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker, mainly to assess the functions of their Shiga toxin effector polypeptide components. The cytotoxicities of exemplary, cell-targeting molecules were determined using cells expressing, at a cellular surface, significant amounts of the appropriate, extracellular target biomolecule, such as, a target of the binding region scFv4. The cells used in this Example were immortalized, human tumor cells available from the ATCC (Manassas VA, U.S.), National Cancer Institute of the U.S. (Frederick, MD, U.S.), and/or DSZM (Braunschweig, DE), such as HCC-1954, MDA-MB-231, Daudi, and U-266 cells, or more simply cell-lines A, B, G, and I, respectively. Certain cell-targeting molecules were tested using cell-kill assays involving both target biomolecule positive and target biomolecule negative cells with respect to the target biomolecule of each cell-targeting molecule's binding region.
[0588] The cytotoxicity assays were performed as follows. Certain, human tumor cell-line cells were plated (typically at 210.sup.3 cells per well for adherent cells, plated the day prior to cell-targeting molecule addition, or 7.510.sup.3 cells per well for suspension cells, plated the same day as cell-targeting molecule addition) in 20 L cell culture medium in 384-well plates. A series of 10-fold dilutions of the molecules to be tested was prepared in an appropriate buffer, and 5 L of the dilutions or buffer control were added to the plated cells. Control wells containing only cell culture medium were used for baseline correction. The cell samples were incubated with the cell-targeting molecule or just buffer for 3 or 5 days at 37 C. and in an atmosphere of 5% carbon dioxide (CO.sub.2). The total cell survival or percent viability was determined using a luminescent readout using the CellTiter-Glo Luminescent Cell Viability Assay or RealTime-Glo MT Cell Viability Assay (Promega Corp., Madison, WI, U.S.) according to the manufacturer's instructions.
[0589] The Percent Viability of experimental wells was calculated using the following equation: (Test RLUAverage Media RLU)(Average Cells RLUAverage Media RLU)100. The logarithm of the cell-targeting molecule protein concentration versus Percent Viability was plotted in Prism (GraphPad Prism, San Diego, CA, U.S.) and log (inhibitor) versus response (3 parameter) analysis or and log (inhibitor) versus normalized response analysis were used to determine the half-maximal cytotoxic concentration (CD.sub.50) value for the tested molecule. The CD.sub.50 value(s) for each molecule tested were calculated, when possible, and shown in Table 9. When CD.sub.50 values could not be calculated based on the shape of the curve over the concentrations tested, then a maximum CD.sub.50 value was noted as being beyond the maximum tested value, e.g., greater than 20,000 nanogram/milliliter (>20,000 ng/mL), for samples which did not kill 50% of the cells at the highest, tested, sample concentration, e.g., 100 or 200 nanomolar (nM). In some experiments, biomolecule target negative cells treated with the maximum concentration of the cell-targeting molecule did not show any change in viability as compared to a buffer only control. The CD.sub.50 values for exemplary, cell-targeting molecules are shown in Table 9 and associated cell-kill assay data is shown in
TABLE-US-00011 TABLE 9 Cytotoxic Activities of Exemplary, Cell-Targeting Molecules SLT-1A Cell-type tested Cytotoxicity Cell-type tested Cytotoxicity lysine (target positive CD.sub.50 (target positive CD.sub.50 Molecule tested position or negative) (ng/mL) or negative) (ng/mL) SLT-1A-Lys1-D3- K1 Cell Line A 1.61 Cell Line G >20,000 variant-1::scFv4 (positive) (negative) SEQ ID NO: 818 SLT-1A-Lys1-D3- K1 Cell Line A 1.89 Cell Line G >20,000 variant-2::scFv4 (positive) (negative) SEQ ID NO: 819 SLT-1A-Lys1-D3- K1 Cell Line A 2.99 Cell Line G >20,000 variant-4::scFv4 (positive) (negative) SEQ ID NO: 820 SLT-1A-Lys2-D3- K11 Cell Line A 0.58 Cell Line G >20,000 variant-1::scFv4 (positive) (negative) SEQ ID NO: 821 SLT-1A-Lys2-D3- K11 Cell Line A 2.40 Cell Line G >20,000 variant-2::scFv4 (positive) (negative) SEQ ID NO: 822 SLT-1A-Lys2-D3- K11 Cell Line A 2.99 Cell Line G >20,000 variant-5::scFv4 (positive) (negative) SEQ ID NO: 823 SLT-1A- K1/K11 Cell Line A 1.22 Cell Line G >20,000 D3::scFv4 control (positive) (negative) SEQ ID NO: 828
[0590] The molecules tested for cytotoxic activity in this Example included cell-targeting molecules comprising or consisting of SLT-1A-Lys1-D3-variant-1::scFv4 (SEQ ID NO:818), SLT-1A-Lys1-D3-variant-2::scFv4 (SEQ ID NO:819), SLT-1A-Lys1-D3-variant4::scFv4 (SEQ ID NO:820), SLT-1A-Lys2-D3-variant-1::scFv4 (SEQ ID NO:821), SLT-1A-Lys2-D3-variant2::scFv4 (SEQ ID NO:822), and SLT-1A-Lys2-D3-variant-5::scFv4 (SEQ ID NO:823). The exemplary cell-targeting molecules of the present invention represented by SEQ ID NOs: 818-823 were each cytotoxic to target positive cells, commonly exhibiting CD.sub.50 values less than 3 ng/mL, depending on the cell-line tested. The cytotoxicities of the Shiga toxin effector polypeptides (SEQ ID NOs: 197, 198, 200-202, and 205) tested as a component of a cell-targeting molecule were comparable to the cytotoxicities of a related, Shiga toxin effector polypeptide having two or more endogenous lysine residues (see e.g. cytotoxicities measured for SLT-1A-D3::scFv4 (SEQ ID NO:828) in Table 9 and
[0591] Similar to the use of cysteine residues in Examples 1-5, a unique lysine residue may be used for site-specific attachment of other molecules using conjugation chemical reactions known to the skilled worker. Commonly, two molecules are linked via an acylation of the functional group of the lysine.
Example 7. Constructing Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention by Removing Lysine Residues Via Amino Acid Residue Substitution
[0592] This example describes the creation of various scaffolds comprising Shiga toxin effector polypeptides, each comprising one, unique, lysine residue. All other lysine residues are removed from the Shiga toxin effector polypeptides as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. Exemplary, cell-targeting molecules of the present invention (e.g. scFv(n)::SLT-1A-Lys(p) and SLT-1A-Lys(p)::scFv(n), where ID numbers the unique lysine residue) are created using these Shiga toxin effector polypeptides.
[0593] The parental, Shiga toxin effector polypeptide of this Example is the A Subunit of Shiga toxin (SEQ ID NO:2), Shiga-like toxin 1 (SEQ ID NO:1), and Shiga-like toxin 2 (SLT-2A) (SEQ ID NO:3). Using standard techniques, the parental, Shiga toxin effector polypeptide is used to make various Shiga toxin effector polypeptides, each having one unique lysine residue (see e.g. Table 10). Cell-targeting molecules (see e.g. SEQ ID NOs: 773-783) comprising Shiga toxin effector polypeptides (see e.g. SEQ ID NOs: 125-232, 1109-1140) having exactly one of the unique lysine residues described in Table 10 are made using standard techniques and tested for retention of Shiga toxin A Subunit function as in Example 1.
TABLE-US-00012 TABLE 10 Shiga Toxin Effector Polypeptides Engineered to have Unique Lysine Residues Exemplary, Shiga Unique Toxin Effector Lysine Sequence Variants Polypeptide Residue (Shiga Toxin A Subunit-Lys(p)-variant) StxA-Lys1 K1 SEQ ID NOs: 1109 and 1112 StxA-Lys2 K11 SEQ ID NOs: 1110 and 1113-1114 StxA-Lys3 K274 SEQ ID NOs: 1111 and 1115-1116 SLT-1A-Lys1 K1 SEQ ID NOs: 1117 and 1120 SLT-1A-Lys2 K11 SEQ ID NOs: 1118 and 1121-1122 SLT-1A-Lys3 K274 SEQ ID NOs: 1119 and 1123-1124 SLT-2A-Lys1 K11 SEQ ID NOs: 1125 and 1130 SLT-2A-Lys2 K255 SEQ ID NOs: 1126, 1131, 1138 and 1135 SLT-2A-Lys3 K257 SEQ ID NOs: 1127, 1132, 1136, 1139 SLT-2A-Lys4 K270 SEQ ID NOs: 1128, 1133, 1137 and 1140 SLT-2A-Lys5 K288 SEQ ID NOs: 1129 and 1134
[0594] Exemplary, cell-targeting molecules of the present invention are tested for retention of Shiga toxin A Subunit functions after removal of endogenous lysine residues from their Shiga toxin effector polypeptide components. The Shiga toxin A Subunit functions analyzed are: catalytic activity, inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol.
[0595] The catalytic activities of Shiga toxin effector polypeptide components of exemplary, cell-targeting molecules of this Example are tested using a ribosome inhibition assay. The ribosome inactivation activities of all the Shiga toxin effector polypeptides SLT-1A-Lys(p) tested in the context of a cell-targeting molecule are equivalent to the catalytic activity of similarly de-immunized Shiga toxin effector polypeptides comprising all the natively occurring lysine residues and/or wild-type, Shiga toxin A1 fragments (amino acids 1-251 of SEQ ID NO:1 or SEQ ID NO:2; or amino acids 1-250 of SEQ ID NO:3).
[0596] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of this Example, which each comprised a Shiga toxin effector polypeptide SLT-1A-Lys(p), are determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker. The exemplary cell-targeting molecules of the present invention are each cytotoxic to target positive cells, commonly exhibiting CD.sub.50 values less than 100 nM, depending on the cell-line tested. The cytotoxicities of many of the Shiga toxin effector polypeptides tested as a component of a cell-targeting molecule are comparable to the cytotoxicities of a related, Shiga toxin effector polypeptide having two or more endogenous lysine residues.
[0597] Exemplary cell-targeting molecules of this Example are produced, purified and conjugated to another molecule via a lysine residue of the Shiga toxin effector polypeptide component of the cell-targeting molecule using routine methods known to the skilled worker, such as, e.g., wherein the other molecule comprises a succinimidyl ester-fluorescent dye or iso/isothio-cyanate-fluorescent dye. These cell-targeting molecules are tested for cell-targeting, internalization, multimerization, and/or cytotoxicity as described in Example 5. The results of cytotoxicity assays show that cargo-linked cell-targeting molecules of this Example retain potent and specific cytotoxicity to target positive cells that is comparable in potency and activity to unconjugated variants of these cell-targeting molecules. The results of the experiments in this Example demonstrate that a cargo molecule can be conjugated to a unique lysine residue of a Shiga toxin effector polypeptide component of a cell-targeting molecule of the present invention, and, furthermore, such conjugated cell-targeting molecules exhibit a useful level of cargo molecule delivery to the interior of a target positive cell and/or cytotoxicity to targeted cells.
Example 8. Constructing Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention by Removing Lysine Residues Via Amino Acid Residue Substitution
[0598] This example describes the creation of various scaffolds comprising Shiga toxin effector polypeptides, each comprising zero lysine residues. All lysine residues respectively are removed from the Shiga toxin effector polypeptides as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. Exemplary, cell-targeting molecules of the present invention (e.g. SLT-1A-Lys(null)::scFv(n) where null refers to the lack of any lysine residue in the Shiga toxin A Subunit component) are created using these Shiga toxin effector polypeptides.
[0599] The parental, Shiga toxin effector polypeptide of this Example is the A Subunit of Shiga toxin (SEQ ID NO:2), Shiga-like toxin 1 (SEQ ID NO:1), and Shiga-like toxin 2 (SLT-2A) (SEQ ID NO:3). Using standard techniques, the parental, Shiga toxin effector polypeptide is used to make various Shiga toxin effector polypeptides, each lacking any lysine residues (see e.g. Table 11). Cell-targeting molecules (see e.g. SEQ ID NOs: 773-783 and 824-827) comprising Shiga toxin effector polypeptides (see e.g. SEQ ID NOs: 233-720) are made using standard techniques and tested for retention of Shiga toxin A Subunit function as in Examples 1 and 6.
TABLE-US-00013 TABLE 11 Shiga Toxin Effector Polypeptides Engineered to have Unique Lysine Residues Exemplary, Shiga Toxin Sequence Variants Effector Polypeptide Lysine Null (Shiga Toxin A Subunit-Lys(null)-variant) SLT-1A-Lys(null) no lysines SEQ ID NOs: 233-252, 314-333, 477-496, and 558-577 SLT-1A-Lys(null)-FR no lysines SEQ ID NOs: 253-272, 334-353, 497-516, and 578-597 SLT-1A-Lys(null)-D1 no lysines SEQ ID NOs: 273-292, 354-373, 517-536, and 598-617 SLT-1A-Lys(null)-D2 no lysines SEQ ID NOs: 293-313, 374-394, 537-557, and 618-638 SLT-1A-Lys(null)-D3 no lysines SEQ ID NOs: 395-414, 436-455, 639-658, and 680-699 SLT-1A-Lys(null)-D4 no lysines SEQ ID NOs: 415-435, 456-476, 659-679, and 700-720
[0600] Exemplary, cell-targeting molecules of the present invention were tested for retention of Shiga toxin A Subunit functions after removal of all lysine residues from their Shiga toxin effector polypeptide components. The Shiga toxin A Subunit functions analyzed are: catalytic activity, inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol.
[0601] The catalytic activities of Shiga toxin effector polypeptide components of exemplary, cell-targeting molecules of this Example are tested using a ribosome inhibition assay. The ribosome inactivation activities of Shiga toxin effector polypeptides StxA-Lys(null) variants, SLT-1A-Lys(null) variants, and SLT-2A-Lys(null) variants are tested in the context of a cell-targeting molecule and found to be equivalent to the catalytic activity of similarly de-immunized Shiga toxin effector polypeptides comprising all the endogenous, natively occurring lysine residues and/or wild-type, Shiga toxin A1 fragments (such as, e.g., amino acids 1-251 of SEQ ID NO:1 or SEQ ID NO:2; or amino acids 1-250 of SEQ ID NO:3).
[0602] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of this Example, which each comprised a Shiga toxin effector polypeptide StxA-Lys(null), SLT-1A-Lys(null), or SLT-2A-Lys(null) are determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker.
[0603] Exemplary, cell-targeting molecules of the present invention were tested for retention of Shiga toxin A Subunit functions as described in Example 6 after removal of all endogenous lysine residues from their Shiga toxin effector polypeptide components. The Shiga toxin A Subunit functions analyzed were: inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol. The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of this Example, which each comprised a Shiga toxin effector polypeptide SLT-1A-Lys(null) variant as described in Table 11, were determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker, mainly to assess the functions of their Shiga toxin effector polypeptide components. The CD.sub.50 values for exemplary, cell-targeting molecules are shown in Table 12 and associated cell-kill assay data is shown in
TABLE-US-00014 TABLE 12 Cytotoxic Activities of Exemplary, Cell-Targeting Molecules SLT-1A Cell-type tested Cytotoxicity Cell-type tested Cytotoxicity Lysine (target positive CD.sub.50 (target positive CD.sub.50 Molecule tested position or negative) (ng/mL) or negative) (ng/mL) SLT-1A- none Cell Line A 1.62 Cell Line G >20,000 Lys(null)-D3- (positive) (negative) variant-21::scFv4 SEQ ID NO: 824 SLT-1A- none Cell Line A 2.4 Cell Line G >20,000 Lys(null)-varaint- (positive) (negative) 27::scFv4 SEQ ID NO: 825 SLT-1A- none Cell Line A 8.81 Cell Line G >20,000 Lys(null)-D3- (positive) (negative) variant-40::scFv4 SEQ ID NO: 826 SLT-1A- none Cell Line A 1.04 Cell Line I >20,000 Lys(null)-D4- (positive) (negative) variant-42::scFv4 SEQ ID NO: 827 SLT-1A- K1/K11 Cell Line A 1.01 Cell Line G >20,000 D3::scFv4 control (positive) (negative) SEQ ID NO: 828 SLT-1A- K1/K11 Cell Line A 1.36 D5::scFv4 control (positive) SEQ ID NO: 829
[0604] The exemplary cell-targeting molecules of the present invention (SEQ ID NOs: 824-827) are each cytotoxic to target positive cells, commonly exhibiting CD.sub.50 values less than 10 ng/mL, depending on the cell-line tested (see Table 12 and
[0605] Exemplary cell-targeting molecules of this Example are produced, purified and conjugated to another molecule via a lysine residue of the cell-targeting molecule using routine methods known to the skilled worker, such as, e.g., wherein the other molecule comprises a succinimidyl ester-fluorescent dye or iso/isothio-cyanate-fluorescent dye. These cell-targeting molecules are tested for cell-targeting, internalization, multimerization, and/or cytotoxicity as described in Example 5. The results of cytotoxicity assays show that cargo-linked cell-targeting molecules of this Example retain potent and specific cytotoxicity to target positive cells that is comparable in potency and activity to unconjugated variants of these cell-targeting molecules. The results of the experiments in this Example demonstrate that a cargo molecule can be conjugated to a free lysine residue of a component of a cell-targeting molecule of the present invention comprising only those Shiga toxin effector polypeptide(s) lacking lysine residues, and, furthermore, such conjugated cell-targeting molecules exhibit a useful level of cargo molecule delivery to the interior of a target positive cell and/or cytotoxicity to targeted cells.
Example 9. Exemplary, Cell-Targeting Molecules of the Present Invention Comprising a Lysine or Free Cysteine for Site-Specific Conjugation Outside the Toxin Effector Region
[0606] This example describes the creation of various scaffolds comprising Shiga toxin effector polypeptides, each lacking either any cysteine residue or any lysine residue. Many of these scaffolds involved a linker or extension consisting of a peptide or polypeptide fused directly to a Shiga toxin A Subunit effector polypeptide lacking any cysteine or lysine residue. As used in this example, a linker represents a structure in a cell-targeting molecule which links a Shiga toxin effector polypeptide to a cell-targeting binding region capable of binding an extracellular target biomolecule with high affinity wherein the linker is neither part of the Shiga toxin effector polypeptide region structure nor part of the cell-targeting binding region structure.
[0607] In this Example, Shiga toxin A Subunit effector polypeptides lacking any cysteine and/or lysine residue (such as, e.g., SLT-1A-Lys(null) variants described in Example 8) were created and fused to cell-targeting moieties. This allows for the creation of cell-targeting molecules comprising such a Shiga toxin A Subunit effector polypeptide wherein there is a unique lysine or free cysteine present outside of the toxin effector region. The removal of all cysteine or lysine residues from the toxin effector region prevents other conjugation sites from being available when certain conjugation reactions are performed on such cell-targeting molecules. A free cysteine or unique lysine residue outside the Shiga toxin component of a cell-targeting molecule of the present invention may be used for site-specific attachment of other molecules using conjugation chemical reactions known to the skilled worker.
[0608] Using routine techniques known to the skilled worker, all cysteine and/or lysine residues are removed from Shiga toxin effector polypeptides by introducing one or more amino acid residue substitutions as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. The parental, Shiga toxin effector polypeptide of this Example is the A Subunit of Shiga toxin (SEQ ID NO:2), Shiga-like toxin 1 (SEQ ID NO:1), and Shiga-like toxin 2 (SLT-2A) (SEQ ID NO:3). The parental, Shiga toxin effector polypeptide is used to make various Shiga toxin effector polypeptides (see e.g. Table 11). Cell-targeting molecules (see e.g. SEQ ID NOs: 773-783 and 824-827) comprising such null Shiga toxin A Subunit effector polypeptides (see e.g. SEQ ID NOs: 233-756) are made using standard techniques and tested for retention of Shiga toxin A Subunit function as in Examples 1 and 6.
[0609] Using standard techniques, Shiga toxin effector polypeptides of this Example were fused to linkers comprising exactly one cysteine or lysine residue to create exemplary Shiga toxin A Subunit effector polypeptide scaffolds of the present invention (also referred to herein as Shiga toxin effector scaffolds). Whichever amino acid residue was absent from the Shiga toxin effector polypeptide (e.g. cysteine or lysine) was included at a single unique position in the Shiga toxin effector scaffold.
[0610] Exemplary Shiga toxin A Subunit effector polypeptide scaffolds of the present invention, each comprising one or more lysine and/or free cysteine residues outside any toxin effector region, were created and tested in the context of a cell-targeting molecule for retention of Shiga toxin A Subunit function as in Examples 1 and 6. In this Example, each exemplary Shiga toxin effector scaffold of the present invention that was tested comprised additional proteinaceous structure outside of the Shiga toxin effector polypeptide region (see Tables 13).
TABLE-US-00015 TABLE 13 Exemplary, Shiga Toxin Effector Polypeptide Scaffolds of the Present Invention Comprising Only One Lysine Residue for Conjugation Toxin Effector Residue Available for Shiga Toxin A Subunit Scaffold Null for Conjugation Scaffold Sequence SLT-1A-Lys(null) cysteine cysteine at position 257 SEQ ID NO: 762 scaffold variant 1 SLT-1A-Lys(null) cysteine cysteine at position 257 SEQ ID NO: 763 scaffold variant 2 SLT-1A-Lys(null) lysine and lysine at position 255 SEQ ID NO: 764 scaffold variant 1 cysteine SLT-1A-Lys(null) lysine and lysine at position 255 SEQ ID NO: 765 scaffold variant 2 cysteine SLT-1A-Lys(null) lysine and lysine at position 255 SEQ ID NO: 766 scaffold variant 3 cysteine SLT-1A-Lys(null) lysine and lysine at position 255 SEQ ID NO: 767 scaffold variant 4 cysteine
[0611] Using standard techniques, exemplary, cell-targeting molecules of the present invention (named e.g. SLT-1A-Lys(null)::scFv(n) or SLT-1A-Cys(null)::scFv(n)), each comprising one or more lysine and/or free cysteine residues outside any toxin effector region, were created using the null Shiga toxin effector polypeptides (e.g. SLT-1A-Lys(null) or SLT-1A-Cys(null), where null represents the lack of any lysine or cysteine residue, respectively). Some of the exemplary cell-targeting molecules of this Example comprises an exemplary, Shiga toxin effector polypeptide scaffolds of this Example (selected from SEQ ID NOs: 762-767, see also Table 13).
[0612] Cell-targeting molecules (see e.g. SEQ ID NOs: 773-783 and 824-827) based on such null Shiga toxin A Subunit effector polypeptides (SEQ ID NOs: 233-756) were made using standard techniques and tested for retention of Shiga toxin A Subunit function as in Examples 1 and 6. In this Example, each cell-targeting molecule that was tested comprised a cell-targeting, immunoglobulin-type, binding region comprising a polypeptide capable of binding to an extracellular target biomolecule with high-affinity (see Tables 13-15). Protein expression and purification of these cell-targeting molecules was performed using standard techniques known to the skilled worker and/or as previously described (see e.g. WO 2014/164680, WO 2014/164693, WO 2015/138435, WO 2015/138452, WO 2015/191764, and WO 2016/126950), such as using Capto-L (GE Healthcare, Marlborough, MA, U.S.).
TABLE-US-00016 TABLE 14 Exemplary Cell-Targeting Molecule Components of the Present Invention Comprising an Engineered, Free Cysteine Available for Conjugation Cell-Targeting Molecule Exemplary, Cell-Targeting Component Conjugation site location Molecules with Component linker-Cys1 between Binding Region and Toxin SEQ ID NOs: 803-806 (SEQ ID NO: 757) Effector Region linker-Cys2 between Binding Region and Toxin (SEQ ID NO: 758) Effector Region linker-Cys3 between Binding Region and Toxin (SEQ ID NO: 759) Effector Region linker-Cys4 between Binding Region and Toxin (SEQ ID NO: 760) Effector Region scFv-linker-Cys1 between the scFv's heavy and light SEQ ID NOs: 807-808 (SEQ ID NO: 769) variable regions scFv-linker-Cys2 between the scFv's heavy and light SEQ ID NOs: 809-811 (SEQ ID NO: 770) variable regions scFv-linker-Cys3 between the scFv's heavy and light (SEQ ID NO: 771) variable regions scFv-linker-Cys4 between the scFv's heavy and light (SEQ ID NO: 772) variable regions scFv(n)-Cys-C2 penultimate C-terminal residue of SEQ ID NOs: 812-815 Binding Region V.sub.HH(n)-Cys-C2 penultimate C-terminal residue of SEQ ID NOs: 816-817 Binding Region
[0613] The cytotoxicities of exemplary, cell-targeting molecules were determined using cells expressing, at a cellular surface, significant amounts of the appropriate, extracellular target biomolecule. Using the cytotoxicity assay, the CD.sub.50 value(s) for each molecule tested were calculated (and reported in Table 15), and associated cell-kill assay data is shown in
TABLE-US-00017 TABLE 15 Cytotoxic Activities of Exemplary, Cell-Targeting Molecules Free Cell-type tested Cytotoxicity Cell-type tested Cytotoxicity Cysteine (target positive CD.sub.50 (target positive CD.sub.50 Molecule Tested location or negative) (nM) or negative) (nM) SLT-1A-D1:: between Toxin Cell Line C 0.0006 Cell Line D 0.0089 linker-Cys1::scFv2 Effector and (positive) (positive) (SEQ ID NO: 803) Binding Region SLT-1A-D1:: between Cell Line C 0.0038 Cell Line D 0.0273 scFv2-linker-Cys1 variable (positive) (positive) (SEQ ID NO: 807) domains of scFv2 SLT-1A-D1:: in variable Cell Line C 0.0002 Cell Line D 0.002 scFv2-Cys-C2 domain of (positive) (positive) (SEQ ID NO: 812) scFv2 SLT-1A-D1::scFv2 none Cell Line C 0.0026 Cell Line D 0.0073 (SEQ ID NO: 838) (positive) (positive) Cell-type tested Cytotoxicity Cell-type tested Cytotoxicity Lysine (target positive CD.sub.50 (target positive CD.sub.50 Molecule Tested location or negative) (ng/mL) or negative) (ng/mL) SLT-1A-Lys(null)- K256 between Cell Line A 1.62 Cell Line G >20,000 D3-variant- Toxin Effector (positive) (negative) 21::scFv4 and Binding (SEQ ID NO: 824) Region SLT-1A-Lys(null)- K256 between Cell Line A 2.4 Cell Line G >20,000 D3-variant- Toxin Effector (positive) (negative) 27::scFv4 and Binding (SEQ ID NO: 825) Region SLT-1A-Lys(null)- K256 between Cell Line A 8.81 Cell Line G >20,000 D3-variant- Toxin Effector (positive) (negative) 40::scFv4 and Binding (SEQ ID NO: 826) Region SLT-1A-D3::scFv4 K1, K11, and Cell Line A 1.22 Cell Line G >20,000 (SEQ ID NO: 828) K256 between (positive) (negative) Toxin Effector and Binding Region SLT-1A-D3::scFv4 K1, K11, and Cell Line A 1.01 Cell Line G >20,000 (SEQ ID NO: 828) K256 between (positive) (negative) Toxin Effector and Binding Region SLT-1A-D3::scFv4 K1, K11 Cell Line A 1.36 variant 2 (positive) (SEQ ID NO: 829)
[0614] Table 15 and
[0615] Table 15 and
Example 10. Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention Comprising a Unique Selenocysteine Residue for Site-Specific Conjugation
[0616] In this Example, exemplary Shiga toxin A Subunit effector polypeptides of the present invention (SLT-1A-Sec(p)-variant), where Sec(p) represents a selenocysteine residue engineered at a unique position), each comprising one ectopic selenocysteine residue, are created and tested as components of exemplary cell-targeting molecules of the present invention. The ectopic, selenocysteine residues are engineered into Shiga toxin effector polypeptides as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. Exemplary, cell-targeting molecules of the present invention (e.g. SLT-1A-Sec(p)-variant::scFv(n), scFv(n)::SLT-1A-Sec(p)-variant), StxA-Sec(p)-variant::scFv(n), and scFv(n)::StxA-Sec(p)-variant) are created using these Shiga toxin effector polypeptides.
[0617] In this Example, cell-targeting molecules, each comprising a Shiga toxin A Subunit effector polypeptide having one ectopic, selenocysteine residue, are created and tested. These cell-targeting molecules each comprised a cell-targeting, immunoglobulin-type, binding region comprising a polypeptide capable of binding to an extracellular target biomolecule with high-affinity.
[0618] The parental, Shiga toxin effector polypeptide of this Example is the A1 fragment of the A Subunit of Shiga toxin (SEQ ID NO:2), Shiga-like toxin 1 (SEQ ID NO:1), and Shiga-like toxin 2 (SLT-2A) (SEQ ID NO:3), optionally comprising the substitution C241S or C242S to remove the only endogenous, cysteine residue in the parental molecule. Using standard techniques, the parental, Shiga toxin effector polypeptide is used to make various Shiga toxin effector polypeptides, each having one unique and ectopic selenocysteine residue and optionally no cysteine residues (see e.g. Table 16). Cell-targeting molecules comprising Shiga toxin effector polypeptides having exactly one of the ectopic selenocysteine residues described in Table 16 are made using standard techniques and tested as described in the Examples above. In Table 16, the code Se-C refers to selenocysteine.
TABLE-US-00018 TABLE 16 Ectopic, Selenocysteine Residues Engineered into Shiga Toxin Effector Polypeptides Exemplary, Shiga toxin effector polypeptides Selenocysteine StxA-Sec(p) and SLT-1A-Sec(p) Substitution StxA-Sec1 and SLT-1A-Sec1 K1Se-C StxA-Sec2 and SLT-1A-Sec2 S8Se-C StxA-Sec3 and SLT-1A-Sec3 S16Se-C StxA-Sec4 and SLT-1A-Sec4 S22Se-C StxA-Sec5 and SLT-1A-Sec5 S33Se-C StxA-Sec6 and SLT-1A-Sec6 S43Se-C StxA-Sec7 or SLT-1A-Sec7 (identical) S45Se-C StxA-Sec8 and SLT-1A-Sec8 V54Se-C StxA-Sec9 and SLT-1A-Sec9 S146Se-C StxA-Sec10 and SLT-1A-Sec10 S186Se-C
[0619] Exemplary, cell-targeting molecules of the present invention are tested for retention of Shiga toxin A Subunit functions after introduction of selenocysteine residues into their Shiga toxin effector polypeptide components. The Shiga toxin A Subunit functions analyzed are: catalytic activity, inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol.
[0620] The catalytic activities of Shiga toxin effector polypeptide components of exemplary, cell-targeting molecules of this Example are tested using a ribosome inhibition assay. The ribosome inactivation activities of all the Shiga toxin effector polypeptides SLT-1A-Sec(p) tested in the context of a cell-targeting molecule are equivalent to the catalytic activity of similarly de-immunized Shiga toxin effector polypeptides comprising all the natively occurring cysteine residues and/or wild-type, Shiga toxin A1 fragments (amino acids 1-251 of SEQ ID NO:1 or SEQ ID NO:2; or amino acids 1-250 of SEQ ID NO:3).
[0621] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of this Example, which each comprised a Shiga toxin effector polypeptide SLT-1A-Sec(p), are determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker. The exemplary cell-targeting molecules of the present invention are each cytotoxic to target positive cells, commonly exhibiting CD.sub.50 values less than 100 nM, depending on the cell-line tested. The cytotoxicities of many of the Shiga toxin effector polypeptides tested as a component of a cell-targeting molecule are comparable to the cytotoxicities of a related, Shiga toxin effector polypeptide having all its endogenous cysteine residues.
[0622] Exemplary cell-targeting molecules of this Example are produced, purified and conjugated to another molecule via a selenocysteine of the cell-targeting molecule using routine methods known to the skilled worker, such as, e.g., wherein the other molecule comprises a succinimidyl ester-fluorescent dye or iso/isothio-cyanate-fluorescent dye. These cell-targeting molecules are tested for cell-targeting, internalization, multimerization, and/or cytotoxicity as described in Example 5. The results of cytotoxicity assays show that cargo-linked cell-targeting molecules of this Example retain potent and specific cytotoxicity to target positive cells that is comparable in potency and activity to unconjugated variants of these cell-targeting molecules. The results of the experiments in this Example demonstrate that a cargo molecule can be conjugated to a selenocysteine residue of a Shiga toxin effector polypeptide component of a cell-targeting molecule of the present invention, and, furthermore, such conjugated cell-targeting molecules exhibit a useful level of cargo molecule delivery to the interior of a target positive cell and/or cytotoxicity to targeted cells.
Example 11. Exemplary, Shiga Toxin Effector Polypeptides of the Present Invention Comprising a Unique Pyrroline-Carboxy-Lysine Residue for Site-Specific Conjugation
[0623] In this Example, exemplary Shiga toxin A Subunit effector polypeptides of the present invention (SLT-1A-Pcl(p)-variant), where Pcl(p) represents a pyrroline-carboxy-lysine residue engineered at a unique position), each comprising one ectopic pyrroline-carboxy-lysine residue, are created and tested as components of exemplary cell-targeting molecules of the present invention. The ectopic, pyrroline-carboxy-lysine residues are engineered into Shiga toxin effector polypeptides as genetically encoded substitutions which did not change the overall number of amino acid residues in the polypeptide. Exemplary, cell-targeting molecules of the present invention (e.g. SLT-1A-Pcl(p)-variant::scFv(n), scFv(n)::SLT-1A-Pcl(p)-variant), StxA-Pcl(p)-variant::scFv(n), and scFv(n)::StxA-Pcl(p)-variant) are created using these Shiga toxin effector polypeptides.
[0624] In this Example, cell-targeting molecules, each comprising a Shiga toxin A Subunit effector polypeptide having one ectopic, pyrroline-carboxy-lysine residue, are created and tested. These cell-targeting molecules each comprised a cell-targeting, immunoglobulin-type, binding region comprising a polypeptide capable of binding to an extracellular target biomolecule with high-affinity.
[0625] The parental, Shiga toxin effector polypeptide of this Example is the A1 fragment of the A Subunit of Shiga toxin (SEQ ID NO:2), Shiga-like toxin 1 (SEQ ID NO:1), and Shiga-like toxin 2 (SLT-2A) (SEQ ID NO:3), optionally comprising the substitution C241S or C242S to remove the only endogenous, cysteine residue of the parental molecule. Using standard techniques, the parental, Shiga toxin effector polypeptide is used to make various Shiga toxin effector polypeptides, each having one unique and ectopic pyrroline-carboxy-lysine residue and optionally no lysine residues (see e.g. Table 17). Cell-targeting molecules comprising Shiga toxin effector polypeptides having exactly one of the ectopic pyrroline-carboxy-lysine residues described in Table 17 are made using standard techniques and tested as described in the Examples above. In Table 17, the code Pcl refers to pyrroline-carboxy-lysine.
TABLE-US-00019 TABLE 17 Ectopic, Pyrroline-Carboxy-Lysine Residues Engineered into Shiga Toxin Effector Polypeptides Shiga Toxin Effector Polypeptide StxA-Pcl(p), SLT-1A-Pcl(p), or Unique Pcl Residue SLT-2A-Pcl(p) Substitution Position StxA-Pcl1 K1Pcl StxA-Pcl2 K11Pcl StxA-Pcl3 K27Pcl SLT-1A-Pcl1 K1Pcl SLT-1A-Pcl2 K11Pcl SLT-1A-Pcl3 K274Pcl SLT-2A-Pcl1 K11Pcl SLT-2A-Pcl2 K255Pcl SLT-2A-Pcl3 K257Pcl SLT-2A-Pcl4 K270Pcl SLT-2A-Pcl5 K288Pcl
[0626] Exemplary, cell-targeting molecules of the present invention are tested for retention of Shiga toxin A Subunit functions after introduction of pyrroline-carboxy-lysine residues into their Shiga toxin effector polypeptide components. The Shiga toxin A Subunit functions analyzed are: catalytic activity, inhibition of eukaryotic ribosome function, cytotoxicity, and by inference self-directing subcellular routing to the cytosol.
[0627] The catalytic activities of Shiga toxin effector polypeptide components of exemplary, cell-targeting molecules of this Example are tested using a ribosome inhibition assay. The ribosome inactivation activities of all the Shiga toxin effector polypeptides SLT-1A-Pcl(p) tested in the context of a cell-targeting molecule are equivalent to the catalytic activity of similarly de-immunized Shiga toxin effector polypeptides comprising all the natively occurring lysine residues and/or wild-type, Shiga toxin A1 fragments (amino acids 1-251 of SEQ ID NO:1 or SEQ ID NO:2; or amino acids 1-250 of SEQ ID NO:3).
[0628] The potency and specificity of cytotoxic activities of exemplary, cell-targeting molecules of this Example, which each comprised a Shiga toxin effector polypeptide SLT-1A-Pcl(p), are determined using a tissue culture cell-based cytotoxicity assay known to the skilled worker. The exemplary cell-targeting molecules of the present invention are each cytotoxic to target positive cells, commonly exhibiting CD.sub.50 values less than 100 nM, depending on the cell-line tested. The cytotoxicities of many of the Shiga toxin effector polypeptides tested as a component of a cell-targeting molecule are comparable to the cytotoxicities of a related, Shiga toxin effector polypeptide comprising all the natively occurring lysine residues and no pyrroline-carboxy-lysine residues or alternatively a wild-type, Shiga toxin A1 fragment.
[0629] Exemplary cell-targeting molecules of this Example are produced, purified and conjugated to another molecule via a pyrroline-carboxy-lysine residue of the cell-targeting molecule using routine methods known to the skilled worker, such as, e.g., wherein the other molecule comprises a succinimidyl ester-fluorescent dye or iso/isothio-cyanate-fluorescent dye. These cell-targeting molecules are tested for cell-targeting, internalization, multimerization, and/or cytotoxicity as described in Example 5. The results of cytotoxicity assays show that cargo-linked cell-targeting molecules of this Example retain potent and specific cytotoxicity to target positive cells that is comparable in potency and activity to unconjugated variants of these cell-targeting molecules. The results of the experiments in this Example demonstrate that a cargo molecule can be conjugated to a pyrroline-carboxy-lysine residue of a Shiga toxin effector polypeptide component of a cell-targeting molecule of the present invention, and, furthermore, such conjugated cell-targeting molecules exhibit a useful level of cargo molecule delivery to the interior of a target positive cell and/or cytotoxicity to targeted cells.
Example 12. Cell-Targeting Molecules Targeting Various Cell Types, Each Comprising a Shiga Toxin a Subunit Effector Polypeptide of the Present Invention
[0630] In this example, cell-targeting molecules of the present invention are created using one or more Shiga toxin effector polypeptides described herein to provide a unique amino acid residue for site-specific attachment of a cargo and/or cell-targeting molecule altering agent wherein the Shiga toxin effector polypeptide(s) also provide two or more of the following: 1) de-immunization, 2) protease-cleavage resistance, and/or 3) an embedded or inserted, heterologous, T-cell epitope. An immunoglobulin-type binding region is derived from the molecules chosen from column 1 of Table 18 and which binds the extracellular target biomolecule indicated in column 2 of Table 18. The selected immunoglobulin-type binding region and Shiga toxin effector polypeptide are associated with each other. The exemplary proteins of this example are optionally created with a carboxy-terminal KDEL-type signal motif (KDEL disclosed as SEQ ID NO:1142) using techniques known in the art and optionally linked to an additional exogenous material, such as, e.g., a useful agent: detection-promoting agent, solubility-altering agent, pharmacokinetic-altering agent, immunogenicity-altering agent, and/or pharmacodynamic-altering agent like a polyethylene glycol or serum albumin, and/or an additional exogenous material: a peptide, protein, nucleic acid, protein-nucleic acid complex, cytotoxic agent, or antibiotic like an antigen, enzyme, or messenger RNA. The resulting molecules are tested for any function required to be categorized as a cell-targeting molecule of the present invention as described in the previous examples using cells expressing the appropriate extracellular target biomolecules.
TABLE-US-00020 TABLE 18 Various Binding Regions for Cell-Targeting Molecules of the Present Invention Source of binding region Extracellular target Application(s) alemtuzumab CD52 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders basiliximab CD25 T-cell disorders, such as prevention of organ transplant rejections, and some B-cell lineage cancers brentuximab CD30 hematological cancers, B-cell related immune disorders, and T-cell related immune disorders catumaxomab EpCAM various cancers, such as ovarian cancer, malignant ascites, gastric cancer cetuximab EGFR various cancers, such as colorectal cancer and head and neck cancer daclizumab CD25 B-cell lineage cancers and T-cell disorders, such as rejection of organ transplants daratumumab CD38 hematological cancers, B-cell related immune disorders, and T-cell related immune disorders dinutuximab ganglioside GD2 Various cancers, such as breast cancer, myeloid cancers, and neuroblastoma efalizumab LFA-1 (CD11a) autoimmune disorders, such as psoriasis enoblituzumab CD276 (B7-H3) various cancers and immune disorders ertumaxomab HER2/neu various cancers and tumors, such as breast cancer and colorectal cancer gemtuzumab CD33 myeloid cancer or immune disorder ibritumomab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders ipilimumab CD152 T-cell related disorders and various cancers, such as leukemia, melanoma muromonab CD3 prevention of organ transplant rejections natalizumab integrin 4 autoimmune disorders, such as multiple sclerosis and Crohn's disease obinutuzumab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders ocaratuzumab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders ocrelizumab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders ofatumumab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders palivizumab F protein of respiratory treat respiratory syncytial virus syncytial virus panitumumab EGFR various cancers, such as colorectal cancer and head and neck cancer pertuzumab HER2/neu various cancers and tumors, such as breast cancer and colorectal cancer pro 140 CCR5 HIV infection and T-cell disorders ramucirumab VEGFR2 various cancers and cancer related disorders, such as solid tumors rituximab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders tocilizumab or IL-6 receptor autoimmune disorders, such as rheumatoid arthritis atlizumab tositumomab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders trastuzumab HER2/neu various cancers and tumors, such as breast cancer and colorectal cancer ublituximab CD20 B-cell cancers, such as lymphoma and leukemia, and B- cell related immune disorders, such as autoimmune disorders vedolizumab integrin 47 autoimmune disorders, such as Crohn's disease and ulcerative colitis CD20 binding CD20 B-cell cancers, such as lymphoma and leukemia, and B- antibodies, cell related immune disorders, such as autoimmune scFv(s), and disorders (see e.g. Geng S et al., Cell Mol Immunol 3: 439- fibronectin 43 (2006); Olafesn T et al., Protein Eng Des Sel 23: 243-9 domain(s) (2010); Natarajan A et al., Clin Cancer Res 19: 6820-9 FN3.sub.CD20 (2013)) CD22 binding CD22 B-cell cancers or B-cell related immune disorders (see e.g. scFv(s) Kawas S et al., MAbs 3: 479-86 (2011)) CD24 binding CD24 various cancers, such as bladder cancer (see e.g. monoclonal Kristiansen G et al., Lab Invest 90: 1102-16 (2010)) antibody(ies) CD25 binding CD25 various cancers of the B-cell lineage and immune scFv(s) disorders related to T-cells (see e.g. Muramatsu H et al., Cancer Lett 225: 225-36 (2005)) CD30 binding CD30 B-cell cancers or B-cell/T-cell related immune disorders monoclonal (see e.g. Klimka A et al., Br J Cancer 83: 252-60 (2000)) antibody(ies) CD33 binding CD33 myeloid cancer or immune disorder (see e.g. Benedict C et monoclonal al., J Immunol Methods 201: 223-31 (1997)) antibody(ies) CD38 binding CD38 hematological cancers, B-cell related immune disorders, immunoglobulin and T-cell related immune disorders (see e.g. U.S. Pat. No. domains 8,153,765) CD40 binding CD40 various cancers and immune disorders (see e.g. Ellmark P scFv(s) et al., Immunology 106: 456-63 (2002)) CD45 binding CD45 Hematological cancers and myelodysplastic syndromes monoclonal (see e.g. Matthews D et al., Blood 94: 1237-47 (1999); Lin antibody(ies) Y et al., Cancer Res 66: 3884-92 (2006); Pagel J et al., and scFv(s) Blood 107: 2184-91 (2006)) CD52 binding CD52 B-cell cancers, such as lymphoma and leukemia, and B- monoclonal cell related immune disorders, such as autoimmune antibody(ies) disorders (see e.g. U.S. Pat. No. 7,910,104) CD56 binding CD56 immune disorders and various cancers, such as lung monoclonal cancer, Merkel cell carcinoma, myeloma (see e.g. Shin J et antibody(ies) al., Hybridoma 18: 521-7 (1999)) CD79 binding CD79 B-cell cancers or B-cell related immune disorders (see e.g. monoclonal Zhang L et al., Ther Immunol 2: 191-202 (1995)) antibody(ies) CD133 binding CD133 various cancers, hematologic malignancies, and immune monoclonal disorders (see e.g. Bidlingmaier S et al., J Mol Med 86: antibodies and 1025-32 (2008); Pavlon L et al., J Microsc 231: 374-83 scFv(s) (2008); Rappa G et al., Stem Cells 26: 3008-17 (2008); Swaminathan S et al., J Immunol Methods 361: 110-5 (2010); Wang J et al., Hybridoma 29: 241-9 (2010); Zhu X et al., Mol Cancer Ther 9: 2131-41 (2010); Xia J et al., Sci Rep 3: 3320 (2013)) CD248 binding CD248 various cancers, such as inhibiting angiogenesis (see e.g. scFv(s) Zhao A et al., J Immunol Methods 363: 221-32 (2011)) CEA-binding CEA various cancers, such as gastrointestinal cancer, pancreatic antibody(ies), cancer, lung cancer, and breast cancer (see e.g. Neumaier scFv(s), and M et al., Cancer Res 50: 2128-34 (1990); Pavoni E et al., engineered, BMC Cancer 6: 4 (2006); Yazaki P et al., Nucl Med Biol fibronectin 35: 151-8 (2008); Zhao J et al., Oncol Res 17: 217-22 domain(s) (2008); Pirie C et al., J Biol Chem 286: 4165-72 (2011)) FN3.sub.CEA EGFR-binding EGFR various cancers (see e.g. GenBank Accession: 3QWQ_B; Adnectin(s) and GenBank Accession: 2KZI_A; U.S. Pat. No. 8,598,113) Affibody(ies) EpCAM binding EpCAM various cancers, such as ovarian cancer, malignant ascites, monoclonal gastric cancer (see e.g. Schanzer J et al., J Immunother 29: antibody(ies) 477-88 (2006)) PSMA binding PSMA prostate cancer (see e.g. Frigerio B et al., Eur J Cancer 49: monoclonal 2223-32 (2013)) antibody(ies) Eph-B2 binding Eph-B2 various cancers such as colorectal cancer and prostate monoclonal cancer (see e.g. Abngozar M et al., Blood 119: 4565-76 antibody(ies) (2012)) Endoglin Endoglin various cancers, such as breast cancer and colorectal binding cancers (see e.g. Vlkel T et al., Biochim Biophys Res monoclonal Acta 1663: 158-66 (2004)) antibody(ies) FAP binding FAP various cancers, such as sarcomas and bone cancers (see monoclonal e.g. Zhang J et al., FASEB J 27: 581-9 (2013)) antibody(ies) HER2-binding HER2/neu various cancers, such as breast cancer and colorectal monoclonal cancer (see e.g. Zahnd C et al., J Mol Biol 369: 1015-28 antibodies, (2007); WO 1993/21319; WO 1994/00136; WO scFvs, V.sub.HHs, 1997/00271; WO 1998/77797; U.S. Pat. No. 5,772,997; U.S. Pat. No. and DARPins 5,783,186; U.S. Pat. No. 5,821,337; U.S. Pat. No. 5,840,525; U.S. Pat. No. 6,949,245; U.S. Pat. No. 7,625,859; US2011/0059090; Goldstein R et al., Eur J Nucl Med Mol Imaging 42: 288-301 (2015)) LewisY antigen LewisY antigens various cancers, such as cervical and uterine cancer (see binding e.g. Power B et al., Protein Sci 12: 734-47 (2003); monoclonal Feridani A et al., Cytometry 71: 361-70 (2007)) antibody(ies) and scFv(s) Neurotensin neurotensin receptors various cancers (see e.g. GenBank Accession: 2P2C_R); receptor binding Ovigne J et al., Neuropeptides 32: 247-56 (1998); Haase C antibodies and et al., Anticancer Res 3527-33 (2006)) DARPin(s) adalimumab TNF- various cancers and immune disorders, such as rheumatoid arthritis, Crohn's Disease, plaque psoriasis, psoriatic arthritis, ankylosing spondylitis, juvenile idiopathic arthritis, hemolytic disease of the newborn afelimomab TNF- various cancers and immune disorders ald518 IL-6 various cancers and immune disorders, such as rheumatoid arthritis anrukinzumab or IL-13 various cancers and immune disorders ima-638 briakinumab IL-12, IL-23 various cancers and immune disorders, such as psoriasis, rheumatoid arthritis, inflammatory bowel diseases, multiple sclerosis brodalumab IL-17 various cancers and immune disorders, such as inflammatory diseases canakinumab IL-1 various cancers and immune disorders, such as rheumatoid arthritis certolizumab TNF- various cancers and immune disorders, such as Crohn's disease fezakinumab IL-22 various cancers and immune disorders, such as rheumatoid arthritis, psoriasis ganitumab IGF-I various cancers golimumab TNF- various cancers and immune disorders, such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis infliximab TNF- various cancers and immune disorders, such as rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, psoriasis. Crohn's disease, ulcerative colitis ixekizumab IL-17A various cancers and immune disorders, such as autoimmune diseases mepolizumab IL-5 various immune disorders and cancers, such as B-cell cancers nerelimomab TNF- various cancers and immune disorders olokizumab IL6 various cancers and immune disorders ozoralizumab TNF- inflammation perakizumab IL-17A various cancers and immune disorders, such as arthritis placulumab human TNF various immune disorders and cancers sarilumab IL6 various cancers and immune disorders, such as rheumatoid arthritis, ankylosing spondylitis siltuximab IL-6 various cancers and immune disorders sirukumab IL-6 various cancers and immune disorders, such as rheumatoid arthritis tabalumab BAFF B-cell cancers ticilimumab or CTLA-4 various cancers tremelimumab tildrakizumab IL23 immunologically mediated inflammatory disorders tnx-650 IL-13 various cancers and immune disorders, such as B-cell cancers tocilizumab or IL-6 receptor various cancers and immune disorders, such as rheumatoid atlizumab arthritis ustekinumab IL-12, IL-23 various cancers and immune disorders, such as multiple sclerosis, psoriasis, psoriatic arthritis Various growth VEGFR, EGFR, FGFR various cancer, such as breast cancer and colon cancer, factors: VEGF, and to inhibit vascularization EGF1, EGF2, FGF Various IL-2R, IL-6R, IL-23R, various immune disorders and cancers cytokines: IL-2, CD80/CD86, IL-6, IL-23, TNFRSF13/TNFRSF17, CCL2, BAFFs, TNFR TNFs, RANKL Various Epstein-Barr virus latent various cancers, viral infections, and immune disorders antibodies membrane protein (see e.g. Chan B et al., In J Cancer 102: 492-8 (2002); (LMP1) Fang C et al., J Immunol Methods 287: 21-30 (2004)); Nichols J et al., J Virol Methods 116: 79-88 (2004); Sim A et al., Sci Rep 3: 3232 (2013)) Broadly Influenza surface viral infections (see e.g. Prabakaran P et al., Front neutralizing antigens (e.g. Microbiol 3: 277 (2012)) antibodies hemaglutinins and identified from matrix protein 2) patient samples Broadly Coronavirus surface viral infections (see e.g. Prabakaran P et al., Front neutralizing antigens Microbiol 3: 277 (2012)) antibodies identified from patient samples Various Filovirus surface viral infections (see e.g. Olinger G et al., Proc Natl Acad antibodies antigens (e.g. VP35, Sci USA 109: 18030-5 (2012); Pettitt J et al., Sci Transl VP40, and glycoprotein) Med 5: 199ra113 (2013); Stahelin R, Expert Opin Ther Targets 18: 115-20 (2014); Becquart P et al., PLoS One 9: e96360 (2014); Stahelin R, Fron Microbiol 5: 300 (2014); Tran E et al., J Virol 88: 10958-62 (2014); Murin C et al., Proc Natl Acad Sci USA 111: 17182-7 (2014)) Broadly Henipavirus surface viral infections (see e.g. Prabakaran P et al., Front neutralizing antigens Microbiol 3: 277 (2012)) antibodies identified from patient samples Various HIV surface antigens viral infections (see e.g. Kitidee K et al., BMC Biotechnol antibodies (e.g. matrix protein 10: 80 (2010); Yu L, Guan Y, Front Immunol 5: 250 including Map17) (2014)) broadly neutralizing antibodies and scFvs
[0631] While some embodiments of the invention have been described by way of illustration, it will be apparent that the invention may be put into practice with many modifications, variations and adaptations, and with the use of numerous equivalents or alternative solutions that are within the scope of persons skilled in the art, without departing from the spirit of the invention or exceeding the scope of the claims.
[0632] All publications, patents, and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. The patent application publications WO 2005/092917, WO 2007/033497, US 2013/196928, WO 2014/164680, WO 2014/164693, WO 2015/113005, WO 2015/113007, WO 2015/120058, WO 2015/138435, WO 2015/138452, US 2015/0259428, WO 2015/191764, US2016/177284, WO 2016/126950, WO 2016/196344, and WO 2017/019623 are each incorporated herein by reference in its entirety. The disclosure of U.S. patent application 62/431,036 is incorporated herein by reference in its entirety. The complete disclosures of all electronically available biological sequence information from GenBank (National Center for Biotechnology Information, U.S.) for amino acid and nucleotide sequences cited herein are each incorporated herein by reference in their entirety.
TABLE-US-00021 SequenceListing IDNumber TextDescription BiologicalSequence SEQIDNO:1 Shiga-liketoxin1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SubunitA(SLT- SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 1A) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVARM ASDEFPSMCPADGRVRGITHNKILWDSSTLGAILMRRTISS SEQIDNO:2 Shigatoxin KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SubunitA(StxA) TGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR MASDEFPSMCPADGRVRGITHNKILWDSSTLGAILMRRTISS SEQIDNO:3 Shiga-liketoxin2 DEFTVDFSSQKSYVDSLNSIRSAISTPLGNISQGGVSVSVINH SubunitA(SLT- VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE 2A) TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ KQKTECQIVGDRAAIKVNNVLWEANTIAALLNRKPQDLTEP NQ SEQIDNO:4 parental,Shiga KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG toxineffector SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN polypeptide NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:5 SLT-1A-Cys1 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:6 SLT-1A-Cys2 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:7 SLT-1A-Cys3 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:8 SLT-1A-Cys4 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:9 SLT-1A-Cys5 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:10 SLT-1A-Cys6 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:11 SLT-1A-Cys7 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG CGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:12 SLT-1A-Cys8 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:13 SLT-1A-Cys9 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLCGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:14 SLT-1A-Cys10 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:15 SLT-1A-Cys1-FR CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:16 SLT-1A-Cys2-FR KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:17 SLT-1A-Cys3-FR KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:18 SLT-1A-Cys4-FR KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:19 SLT-1A-Cys5-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:20 SLT-1A-Cys6-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:21 SLT-1A-Cys7-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG CGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:22 SLT-1A-Cys8-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:23 SLT-1A-Cys9-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLCGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:24 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG FR SGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:25 SLT-1A-Cys1-D1 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:26 SLT-1A-Cys2-D1 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:27 SLT-1A-Cys3-D1 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:28 SLT-1A-Cys4-D1 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:29 SLT-1A-Cys5-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:30 SLT-1A-Cys6-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:31 SLT-1A-Cys7-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:32 SLT-1A-Cys8-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:33 SLT-1A-Cys9-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:34 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D1 IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:35 SLT-1A-Cys1-D2 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:36 SLT-1A-Cys2-D2 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:37 SLT-1A-Cys3-D2 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:38 SLT-1A-Cys4-D2 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:39 SLT-1A-Cys5-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:40 SLT-1A-Cys6-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:41 SLT-1A-Cys7-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:42 SLT-1A-Cys8-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:43 SLT-1A-Cys9-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:44 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D2 IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:45 SLT-1A-Cys1 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:46 SLT-1A-Cys2 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:47 SLT-1A-Cys3 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:48 SLT-1A-Cys4 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:49 SLT-1A-Cys5 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:50 SLT-1A-Cys6 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:51 SLT-1A-Cys7 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) CGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:52 SLT-1A-Cys8 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:53 SLT-1A-Cys9 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:54 SLT-1A-Cys10 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:55 SLT-1A-Cys1-FR CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:56 SLT-1A-Cys2-FR KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:57 SLT-1A-Cys3-FR KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:58 SLT-1A-Cys4-FR KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:59 SLT-1A-Cys5-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:60 SLT-1A-Cys6-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:61 SLT-1A-Cys7-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) CGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:62 SLT-1A-Cys8-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:63 SLT-1A-Cys9-FR KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:64 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG FR(inactivated) SGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:65 SLT-1A-Cys1-D1 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:66 SLT-1A-Cys2-D1 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:67 SLT-1A-Cys3-D1 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:68 SLT-1A-Cys4-D1 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:69 SLT-1A-Cys5-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:70 SLT-1A-Cys6-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:71 SLT-1A-Cys7-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:72 SLT-1A-Cys8-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:73 SLT-1A-Cys9-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:74 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D1(inactivated) IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:75 SLT-1A-Cys1-D2 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:76 SLT-1A-Cys2-D2 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:77 SLT-1A-Cys3-D2 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:78 SLT-1A-Cys4-D2 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:79 SLT-1A-Cys5-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:80 SLT-1A-Cys6-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:81 SLT-1A-Cys7-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:82 SLT-1A-Cys8-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:83 SLT-1A-Cys9-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:84 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D2(inactivated) IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:85 SLT-1A-Cys1-D3 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:86 SLT-1A-Cys2-D3 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:87 SLT-1A-Cys3-D3 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:88 SLT-1A-Cys4-D3 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:89 SLT-1A-Cys5-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:90 SLT-1A-Cys6-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:91 SLT-1A-Cys7-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:92 SLT-1A-Cys8-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:93 SLT-1A-Cys9-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:94 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D3 IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:95 SLT-1A-Cys1-D4 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:96 SLT-1A-Cys2-D4 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:97 SLT-1A-Cys3-D4 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:98 SLT-1A-Cys4-D4 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:99 SLT-1A-Cys5-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:100 SLT-1A-Cys6-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:101 SLT-1A-Cys7-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:102 SLT-1A-Cys8-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:103 SLT-1A-Cys9-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:104 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D4 IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:105 SLT-1A-Cys1-D3 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:106 SLT-1A-Cys2-D3 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:107 SLT-1A-Cys3-D3 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:108 SLT-1A-Cys4-D3 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:109 SLT-1A-Cys5-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:110 SLT-1A-Cys6-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:111 SLT-1A-Cys7-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:112 SLT-1A-Cys8-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:113 SLT-1A-Cys9-D3 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:114 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D3(inactivated) IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:115 SLT-1A-Cys1-D4 CEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:116 SLT-1A-Cys2-D4 KEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:117 SLT-1A-Cys3-D4 KEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:118 SLT-1A-Cys4-D4 KEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:119 SLT-1A-Cys5-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:120 SLT-1A-Cys6-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDCG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:121 SLT-1A-Cys7-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:122 SLT-1A-Cys8-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:123 SLT-1A-Cys9-D4 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:124 SLT-1A-Cys10- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG D4(inactivated) IGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:125 SLT-1A-Lys1 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:126 SLT-1A-Lys1 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:127 SLT-1A-Lys1 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:128 SLT-1A-Lys1 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:129 SLT-1A-Lys2 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:130 SLT-1A-Lys2 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:131 SLT-1A-Lys2 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:132 SLT-1A-Lys2 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:133 SLT-1A-Lys2 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:134 SLT-1A-Lys1-FR KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:135 SLT-1A-Lys1-FR KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:136 SLT-1A-Lys1-FR KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:137 SLT-1A-Lys1-FR KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:138 SLT-1A-Lys2-FR REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:139 SLT-1A-Lys2-FR HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:140 SLT-1A-Lys2-FR DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:141 SLT-1A-Lys2-FR QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:142 SLT-1A-Lys2-FR SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:143 SLT-1A-Lys1-D1 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:144 SLT-1A-Lys1-D1 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:145 SLT-1A-Lys1-D1 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:146 SLT-1A-Lys1-D1 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:147 SLT-1A-Lys2-D1 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:148 SLT-1A-Lys2-D1 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:149 SLT-1A-Lys2-D1 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:150 SLT-1A-Lys2-D1 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:151 SLT-1A-Lys2-D1 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:152 SLT-1A-Lys1-D2 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:153 SLT-1A-Lys1-D2 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:154 SLT-1A-Lys1-D2 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:155 SLT-1A-Lys1-D2 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:156 SLT-1A-Lys2-D2 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:157 SLT-1A-Lys2-D2 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:158 SLT-1A-Lys2-D2 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:159 SLT-1A-Lys2-D2 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:160 SLT-1A-Lys2-D2 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:161 SLT-1A-Lys1 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:162 SLT-1A-Lys1 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:163 SLT-1A-Lys1 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:164 SLT-1A-Lys1 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:165 SLT-1A-Lys2 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:166 SLT-1A-Lys2 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:167 SLT-1A-Lys2 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:168 SLT-1A-Lys2 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:169 SLT-1A-Lys2 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:170 SLT-1A-Lys1-FR KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:171 SLT-1A-Lys1-FR KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:172 SLT-1A-Lys1-FR KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:173 SLT-1A-Lys1-FR KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:174 SLT-1A-Lys2-FR REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:175 SLT-1A-Lys2-FR HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:176 SLT-1A-Lys2-FR DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:177 SLT-1A-Lys2-FR QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:178 SLT-1A-Lys2-FR SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:179 SLT-1A-Lys1-D1 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:180 SLT-1A-Lys1-D1 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:181 SLT-1A-Lys1-D1 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:182 SLT-1A-Lys1-D1 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:183 SLT-1A-Lys2-D1 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:184 SLT-1A-Lys2-D1 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:185 SLT-1A-Lys2-D1 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:186 SLT-1A-Lys2-D1 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:187 SLT-1A-Lys2-D1 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:188 SLT-1A-Lys1-D2 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:189 SLT-1A-Lys1-D2 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:190 SLT-1A-Lys1-D2 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:191 SLT-1A-Lys1-D2 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:192 SLT-1A-Lys2-D2 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:193 SLT-1A-Lys2-D2 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:194 SLT-1A-Lys2-D2 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:195 SLT-1A-Lys2-D2 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:196 SLT-1A-Lys2-D2 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:197 SLT-1A-Lys1-D3 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:198 SLT-1A-Lys1-D3 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:199 SLT-1A-Lys1-D3 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:200 SLT-1A-Lys1-D3 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:201 SLT-1A-Lys2-D3 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:202 SLT-1A-Lys2-D3 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:203 SLT-1A-Lys2-D3 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:204 SLT-1A-Lys2-D3 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:205 SLT-1A-Lys2-D3 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:206 SLT-1A-Lys1-D4 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:207 SLT-1A-Lys1-D4 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:208 SLT-1A-Lys1-D4 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:209 SLT-1A-Lys1-D4 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:210 SLT-1A-Lys2-D4 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:211 SLT-1A-Lys2-D4 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:212 SLT-1A-Lys2-D4 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:213 SLT-1A-Lys2-D4 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:214 SLT-1A-Lys2-D4 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:215 SLT-1A-Lys1-D3 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:216 SLT-1A-Lys1-D3 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:217 SLT-1A-Lys1-D3 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:218 SLT-1A-Lys1-D3 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:219 SLT-1A-Lys2-D3 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:220 SLT-1A-Lys2-D3 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:221 SLT-1A-Lys2-D3 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:222 SLT-1A-Lys2-D3 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:223 SLT-1A-Lys2-D3 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:224 SLT-1A-Lys1-D4 KEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:225 SLT-1A-Lys1-D4 KEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:226 SLT-1A-Lys1-D4 KEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:227 SLT-1A-Lys1-D4 KEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:228 SLT-1A-Lys2-D4 REFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:229 SLT-1A-Lys2-D4 HEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:230 SLT-1A-Lys2-D4 DEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:231 SLT-1A-Lys2-D4 QEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:232 SLT-1A-Lys2-D4 SEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG variant5 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:233 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 1 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:234 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 2 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:235 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:236 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 4 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:237 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 5 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:238 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 6 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:239 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 7 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:240 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 8 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:241 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 9 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:242 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 10 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:243 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 11 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:244 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 12 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:245 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 13 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:246 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 14 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:247 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 15 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:248 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 16 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:249 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 17 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:250 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 18 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:251 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 19 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:252 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:253 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:254 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:255 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:256 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:257 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:258 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:259 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:260 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:261 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:262 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:263 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:264 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:265 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:266 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:267 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:268 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:269 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:270 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:271 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant19 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:272 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:273 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:274 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:275 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:276 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:277 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:278 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:279 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:280 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:281 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:282 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:283 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:284 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:285 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:286 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:287 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:288 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:289 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:290 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:291 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant19 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:292 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D1 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant20 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:293 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:294 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:295 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:296 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:297 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:298 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:299 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:300 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:301 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:302 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:303 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:304 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:305 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:306 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:307 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:308 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:309 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:310 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:311 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D2 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant19 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:312 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:313 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:314 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 1(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:315 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 2(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:316 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 3(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:317 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 4(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:318 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 5(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:319 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 6(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:320 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 7(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:321 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 8(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:322 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 9(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:323 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 10(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:324 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 11(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:325 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 12(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:326 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 13(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:327 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 14(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:328 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 15(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:329 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 16(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:330 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 17(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:331 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 18(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:332 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 19(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:333 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 20(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:334 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:335 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:336 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:337 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:338 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:339 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:340 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:341 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:342 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:343 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:344 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:345 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:346 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:347 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:348 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:349 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:350 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:351 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:352 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant19 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:353 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:354 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:355 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:356 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:357 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:358 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:359 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:360 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:361 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:362 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:363 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:364 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:365 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:366 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:367 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:368 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:369 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:370 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:371 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:372 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant19 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:373 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D1 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant20 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:374 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:375 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:376 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:377 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:378 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:379 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:380 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:381 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:382 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:383 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:384 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:385 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:386 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:387 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:388 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:389 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:390 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:391 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:392 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D2 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant19 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:393 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:394 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:395 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:396 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:397 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:398 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:399 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:400 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:401 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:402 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:403 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:404 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:405 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:406 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:407 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:408 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:409 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:410 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:411 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:412 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:413 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant19 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:414 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D3 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant20 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:415 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:416 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:417 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:418 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:419 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:420 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:421 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:422 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:423 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:424 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:425 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:426 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:427 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:428 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:429 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:430 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:431 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:432 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:433 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D4 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant19 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:434 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:435 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:436 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:437 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:438 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:439 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:440 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:441 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:442 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:443 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:444 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:445 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:446 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:447 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:448 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:449 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:450 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:451 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:452 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:453 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:454 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant19 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:455 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D3 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant20 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:456 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:457 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:458 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:459 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant4 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:460 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant5 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:461 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant6 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:462 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant7 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:463 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant8 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:464 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant9 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:465 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant10 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:466 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant11 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:467 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant12 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:468 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant13 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:469 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant14 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:470 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant15 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:471 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant16 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:472 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant17 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:473 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant18 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:474 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D4 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant19 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:475 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant20 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:476 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:477 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 21 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:478 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 22 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:479 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 23 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:480 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 24 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:481 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 25 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:482 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 26 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:483 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 27 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:484 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 28 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:485 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 29 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:486 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 30 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:487 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 31 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:488 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 32 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:489 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 33 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:490 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 34 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:491 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 35 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:492 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 36 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:493 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 37 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:494 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 38 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:495 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 39 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:496 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 40 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:497 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:498 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:499 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:500 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:501 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:502 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:503 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:504 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:505 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:506 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:507 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:508 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:509 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:510 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:511 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:512 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:513 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:514 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:515 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:516 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant40 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:517 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:518 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:519 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:520 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:521 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:522 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:523 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:524 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:525 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:526 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:527 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:528 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:529 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:530 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:531 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:532 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:533 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:534 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:535 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:536 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D1 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:537 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:538 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:539 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:540 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:541 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:542 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:543 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:544 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:545 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:546 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:547 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:548 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:549 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:550 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:551 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:552 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:553 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:554 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:555 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D2 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:556 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant41 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:557 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant42 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:558 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 21(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:559 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 22(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:560 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 23(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:561 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 24(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:562 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 25(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:563 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 26(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:564 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 27(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:565 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 28(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:566 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 29(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:567 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 30(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:568 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 31(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:569 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 32(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:570 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 33(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:571 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 34(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:572 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 35(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:573 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 36(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:574 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 37(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:575 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 38(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:576 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 39(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:577 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 40(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:578 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:579 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:580 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:581 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:582 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:583 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:584 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:585 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:586 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:587 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:588 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:589 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:590 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:591 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:592 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:593 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:594 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:595 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:596 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:597 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant40 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:598 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:599 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:600 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:601 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:602 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:603 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:604 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:605 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:606 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:607 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:608 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:609 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:610 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:611 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:612 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:613 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:614 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:615 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:616 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:617 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D1 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:618 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:619 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:620 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:621 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:622 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:623 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:624 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:625 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:626 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:627 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:628 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:629 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:630 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:631 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:632 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:633 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:634 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:635 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:636 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D2 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:637 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant41 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:638 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant42 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:639 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:640 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:641 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:642 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:643 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:644 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:645 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:646 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:647 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:648 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:649 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:650 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:651 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:652 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:653 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:654 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:655 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:656 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:657 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:658 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D3 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:659 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:660 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:661 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:662 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:663 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:664 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:665 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:666 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:667 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:668 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:669 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:670 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:671 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:672 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:673 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:674 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:675 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:676 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:677 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D4 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:678 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant41 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:679 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant42 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:680 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant21 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:681 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:682 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:683 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:684 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:685 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:686 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:687 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:688 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:689 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:690 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:691 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:692 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:693 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:694 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:695 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:696 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:697 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:698 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:699 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D3 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:700 SLT-1A- HEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant22 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:701 SLT-1A- DEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant23 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:702 SLT-1A- QEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant24 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:703 SLT-1A- SEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant25 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:704 SLT-1A- REFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant26 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:705 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant27 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:706 SLT-1A- DEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant28 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:707 SLT-1A- QEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant29 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:708 SLT-1A- SEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant30 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:709 SLT-1A- REFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant31 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:710 SLT-1A- HEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant32 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:711 SLT-1A- DEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant33 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:712 SLT-1A- QEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant34 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:713 SLT-1A- SEFTLDFSTAQTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant35 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:714 SLT-1A- REFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant36 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:715 SLT-1A- HEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant37 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:716 SLT-1A- DEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant38 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:717 SLT-1A- QEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant39 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:718 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-D4 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN variant40 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ (inactivated) INRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:719 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant41 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:720 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant42 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:721 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 1 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:722 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:723 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:724 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:725 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 1(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:726 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:727 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:728 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:729 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:730 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:731 SLT-1A- kEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Cys(null)-D3 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN (inactivated) VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ variant1 INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:732 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant1 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:733 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 2 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASRVAR SEQIDNO:734 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASAVAA SEQIDNO:735 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASRVAR SEQIDNO:736 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASRVAR SEQIDNO:737 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN 2(inactivated) NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASRVAR SEQIDNO:738 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASAVAA SEQIDNO:739 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASRVAR SEQIDNO:740 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASRVAR SEQIDNO:741 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASAVAA SEQIDNO:742 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASAVAA SEQIDNO:743 SLT-1A- kEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Cys(null)-D3 GDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTNN (inactivated) VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ variant2 INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNAHHHASAVAA SEQIDNO:744 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNAHHHASAVAA SEQIDNO:745 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)variant SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN 3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASRVAR SEQIDNO:746 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASRVAR SEQIDNO:747 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D1 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASAVAA SEQIDNO:748 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASRVAR SEQIDNO:749 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)variant IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN 3(inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASRVAR SEQIDNO:750 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-FR SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASAVAA SEQIDNO:751 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D1 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASRVAR SEQIDNO:752 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D2 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASRVAR SEQIDNO:753 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASAVAA SEQIDNO:754 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASAVAA SEQIDNO:755 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D3 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN (inactivated) NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM variant3 QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASAVAA SEQIDNO:756 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-D4 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRTN variant3 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM (inactivated) QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNVHHHASAVAA SEQIDNO:757 linker-Cys1 EFPKPCTPPGSSGGAP SEQIDNO:758 linker-Cys2 EFPKPSTPPGSCGGAP SEQIDNO:759 linker-Cys3 AHHCEDPSSKAPKAP SEQIDNO:760 linker-Cys4 AHHSEDPSCKAPKAP SEQIDNO:761 linker-Lys1 EFPKPSTPPGSSGGAP SEQIDNO:762 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-scaffold IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1C257 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAAE FPKPCTPPGSSGGAP SEQIDNO:763 SLT-1A- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Cys(null)-scaffold IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2C257 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAAE FPKPCTPPGSSGGAP SEQIDNO:764 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-scaffold IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant1K255 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAAE FPKPSTPPGSSGGAP SEQIDNO:765 SLT-1A- HEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-scaffold IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2K255 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAAE FPKPSTPPGSSGGAP SEQIDNO:766 SLT-1A- SEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGI Lys(null)-scaffold GDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTNN variant3K255 VFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGMQ INRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRF RQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVLPD YHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAAEFP KPSTPPGSSGGAP SEQIDNO:767 SLT-1A- REFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG Lys(null)-scaffold IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant4K255 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAAE FPKPSTPPGSSGGAP SEQIDNO:768 linker-IgAhinge SPSTPPTPSPSTPPAS SEQIDNO:769 scFv-linker-Cys1 GGGGC SEQIDNO:770 scFv-linker-Cys2 GGGGSGGGGSGGGGCGGGGSGGGGS SEQIDNO:771 scFv-linker-Cys3 GSTSGCGKPGSGEGS SEQIDNO:772 scFv-linker-Cys4 GSTSGCGKPGSGEGSTKG SEQIDNO:773 cell-targeting MKEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule1 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:774 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule2 CGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:775 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule3 SGCGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:776 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule4 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:777 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule5 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:778 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMID molecule6 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIVLTQSPASLAVSLGQRATISCRAT ESVEYYGTSLVQWYQQKPGQPPKLLIYAASSVDSGVPARFS GSGSGTDFSLTIHPVEEDDIAMYFCQQSRRVPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKP GASVKMSCKASGYTFTSYVMEIWVKQKPGQGLEWIGYVNP FNDGTKYNEMFKGKATLTSDKSSSTAYMELSSLTSEDSAVY YCARQAWGYPWGQGTLVTVSA SEQIDNO:779 cell-targeting MKEFTLDFSTAKTYVDCLNVIRSAIGTPLQTISSGGTSLLMID molecule7 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIVLTQSPASLAVSLGQRATISCRAT ESVEYYGTSLVQWYQQKPGQPPKLLIYAASSVDSGVPARFS GSGSGTDFSLTIHPVEEDDIAMYFCQQSRRVPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKP GASVKMSCKASGYTFTSYVMEIWVKQKPGQGLEWIGYVNP FNDGTKYNEMFKGKATLTSDKSSSTAYMELSSLTSEDSAVY YCARQAWGYPWGQGTLVTVSA SEQIDNO:780 cell-targeting MKEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule8 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIVLTQSPASLAVSLGQRATISCRAT ESVEYYGTSLVQWYQQKPGQPPKLLIYAASSVDSGVPARFS GSGSGTDFSLTIHPVEEDDIAMYFCQQSRRVPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKP GASVKMSCKASGYTFTSYVMEIWVKQKPGQGLEWIGYVNP FNDGTKYNEMFKGKATLTSDKSSSTAYMELSSLTSEDSAVY YCARQAWGYPWGQGTLVTVSA SEQIDNO:781 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMID molecule9 SGSGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITVRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRS GTDFTLTISSLQPEDFATYYAQQHYTTPPTFGQGTKVEIKRTG STSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSVAASGFN IKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFT ISADTSKNTAYLQMNSLRAEDTAVYYASRWGGDGFYAMDV WGQGTLVTVSSA SEQIDNO:782 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule10 SGCGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVN RTNNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISR TGMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTA EALRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLS SVLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAV AAEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITVRA SQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSR SGTDFTLTISSLQPEDFATYYAQQHYTTPPTFGQGTKVEIKRT GSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSVAASGF NIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRF TISADTSKNTAYLQMNSLRAEDTAVYYASRWGGDGFYAMD VWGQGTLVTVSSA SEQIDNO:783 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule11 SGSGDNLFAVDCRGIDPEEGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITVRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRS GTDFTLTISSLQPEDFATYYAQQHYTTPPTFGQGTKVEIKRTG STSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSVAASGFN IKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFT ISADTSKNTAYLQMNSLRAEDTAVYYASRWGGDGFYAMDV WGQGTLVTVSSA SEQIDNO:784 cell-targeting MEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG molecule12 QAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV YYCQQRSDWPLTFGGGTKVEIKGSTSGSGKPGSGEGSAVQL VESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKG LEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSSEF PKPSTPPGSSGGAPKEFTLDFCTAKTYVDSLNVIRSAIGTPLQ TISSGGTSLLMIDSGIGDNLFAVDILGFDFTLGRFNNLRLIVER NNLYVTGFVNRTNNVFYRFADFSHVTFPGTTAVTLSADSSY TTLQRVAGISRTGMQINRHSLTTSYLDLMSHSATSLTQSVAR AMLRFVTVTAEALRFRQIQRGFRTTLDDLSGASYVMTAEDV DLTLNWGRLSSVLPDYHGQDSVRVGRISFGSINAILGSVALIL NSHHHASRVAR SEQIDNO:785 cell-targeting MEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG molecule13 QAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV YYCQQRSDWPLTFGGGTKVEIKGSTSGSGKPGSGEGSAVQL VESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKG LEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSSEF PKPSTPPGSSGGAPKEFTLDFSTAKTYVDSLNVIRSAIGTPLQ TISSGGTSLLMIDCGIGDNLFAVDILGFDFTLGRFNNLRLIVER NNLYVTGFVNRTNNVFYRFADFSHVTFPGTTAVTLSADSSY TTLQRVAGISRTGMQINRHSLTTSYLDLMSHSATSLTQSVAR AMLRFVTVTAEALRFRQIQRGFRTTLDDLSGASYVMTAEDV DLTLNWGRLSSVLPDYHGQDSVRVGRISFGSINAILGSVALIL NSHHHASRVAR SEQIDNO:786 cell-targeting MEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG molecule14 QAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV YYCQQRSDWPLTFGGGTKVEIKGSTSGSGKPGSGEGSAVQL VESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKG LEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSSEF PKPSTPPGSSGGAPKEFTLDFSTAKTYVDSLNVIRSAIGTPLQ TISSGGTSLLMIDSGIGDNLFAVDILGFDFTLGRFNNLRLIVER NNLYVTGFVNRTNNVFYRFADFSHVTFPGTTAVTLSADSSY TTLQRVAGISRTGMQINRHSLTTSYLDLMSHCATSLTQSVAR AMLRFVTVTAEALRFRQIQRGFRTTLDDLSGASYVMTAEDV DLTLNWGRLSSVLPDYHGQDSVRVGRISFGSINAILGSVALIL NSHHHASRVAR SEQIDNO:787 cell-targeting MEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG molecule15 QAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV YYCQQRSDWPLTFGGGTKVEIKGSTSGSGKPGSGEGSAVQL VESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKG LEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSSEF PKPSTPPGSSGGAPKEFTLDFSTAKTYVDSLNVIRSAIGTPLQ TISSGGTSLLMIDSGIGDNLFAVDILGFDFTLGRFNNLRLIVER NNLYVTGFVNRTNNVFYRFADFSHVTFPGTTAVTLSADSSY TTLQRVAGISRTGMQINRHSLTTSYLDLMSHSATSLTQSVAR AMLRFVTVTAEALRFRQIQRGFRTTLDDLCGASYVMTAEDV DLTLNWGRLSSVLPDYHGQDSVRVGRISFGSINAILGSVALIL NSHHHASRVAR SEQIDNO:788 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule16 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AAHHSEDPSSKAPKAPEVQLVESGGGLVQAGGSLRLSCAAS GITFSINTMGWYRQAPGKQRELVALISSIGDTYYADSVKGRF TISRDNAKNTVYLQMNSLKPEDTAVYYCKRFRTAAQGTDY WGQGTQVTVSSA SEQIDNO:789 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMID molecule17 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:790 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule18 CGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:791 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule19 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:792 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule20 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:793 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule21 SGCGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:794 cell-targeting MKEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule22 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:795 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule23 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:796 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule24 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:797 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule25 SGCGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:798 cell-targeting MKEFTLDFCTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule26 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:799 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule27 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:800 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule28 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:801 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule29 SGCGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:802 cell-targeting MKEFTLDFSTAKTYVDSLNVIRCAIGTPLQTISSGGTSLLMID molecule30 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:803 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule31 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPCTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:804 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule32 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPCTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:805 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule33 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPCTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:806 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule34 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPCTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:807 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule35 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GCQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:808 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule36 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GCQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:809 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule37 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGCGGGGSGGGGSQVQLQESGPGLVRPSQTL SLTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTD YNAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKS MITTGFVMDSWGQGSLVTVSS SEQIDNO:810 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule38 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGCGGGGSGGGGSQVQLQESGPGLVRPSQTL SLTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTD YNAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKS MITTGFVMDSWGQGSLVTVSS SEQIDNO:811 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule39 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGCGGGGSGGGGSQVQLQESGPGLVRPSQTL SLTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTD YNAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKS MITTGFVMDSWGQGSLVTVSS SEQIDNO:812 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule40 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVCS SEQIDNO:813 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule41 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVCS SEQIDNO:814 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule42 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVCS SEQIDNO:815 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule43 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVCS SEQIDNO:816 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule44 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AAHHSEDPSSKAPKAPEVQLVESGGGLVQAGGSLRLSCAAS GITFSINTMGWYRQAPGKQRELVALISSIGDTYYADSVKGRF TISRDNAKNTVYLQMNSLKPEDTAVYYCKRFRTAAQGTDY WGQGTQVTVSCA SEQIDNO:817 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule45 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLSVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADA LRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA AHHSEDPSSKAPKAPEVQLVESGGGLVQAGGSLRLSCAASGI TFSINTMGWYRQAPGKQRELVALISSIGDTYYADSVKGRFTI SRDNAKNTVYLQMNSLKPEDTAVYYCKRFRTAAQGTDYW GQGTQVTVSCA SEQIDNO:818 cell-targeting MKEFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule46 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKAS GYPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDF KGRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGY VPYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGS DIQMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPG QSPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLA VYFCQQHFRTPFTFGSGTKLEIK SEQIDNO:819 cell-targeting MKEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule47 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKAS GYPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDF KGRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGY VPYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGS DIQMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPG QSPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLA VYFCQQHFRTPFTFGSGTKLEIK SEQIDNO:820 cell-targeting MKEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDS molecule48 GIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA EFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKASG YPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDFK GRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGYV PYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGSDI QMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPGQ SPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLAV YFCQQHFRTPFTFGSGTKLEIK SEQIDNO:821 cell-targeting MREFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule49 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKAS GYPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDF KGRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGY VPYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGS DIQMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPG QSPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLA VYFCQQHFRTPFTFGSGTKLEIK SEQIDNO:822 cell-targeting MHEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule50 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKAS GYPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDF KGRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGY VPYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGS DIQMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPG QSPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLA VYFCQQHFRTPFTFGSGTKLEIK SEQIDNO:823 cell-targeting MSEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDS molecule51 GIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA EFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKASG YPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDFK GRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGYV PYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGSDI QMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPGQ SPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLAV YFCQQHFRTPFTFGSGTKLEIK SEQIDNO:824 cell-targeting MREFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDS molecule52 GIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA EFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKASG YPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDFK GRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGYV PYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGSDI QMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPGQ SPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLAV YFCQQHFRTPFTFGSGTKLEIK SEQIDNO:825 cell-targeting MHEFTLDFSTAHTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule53 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKAS GYPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDF KGRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGY VPYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGS DIQMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPG QSPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLA VYFCQQHFRTPFTFGSGTKLEIK SEQIDNO:826 cell-targeting MSEFTLDFSTASTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDS molecule54 GIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA EFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKASG YPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDFK GRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGYV PYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGSDI QMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPGQ SPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLAV YFCQQHFRTPFTFGSGTKLEIK SEQIDNO:827 cell-targeting MREFTLDFSTARTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDS molecule55 GIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTG MQINRHSLTTSYLALMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SPSTPPTPSPSTPPASQVQLQQSGPELKKPGETVKISCKASGY PFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDFKG RFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGYVP YWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGSDIQ MTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPGQSP KLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLAVYF CQQHFRTPFTFGSGTKLEIK SEQIDNO:828 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule56 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPQVQLQQSGPELKKPGETVKISCKAS GYPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDF KGRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGY VPYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGS DIQMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPG QSPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLA VYFCQQHFRTPFTFGSGTKLEIK SEQIDNO:829 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID molecule57 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA ASPSTPPTPSPSTPPASQVQLQQSGPELKKPGETVKISCKASG YPFTNYGMNWVKQAPGQGLKWMGWINTSTGESTFADDFK GRFDFSLETSANTAYLQINNLKSEDSATYFCARWEVYHGYV PYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSGGGGSDI QMTQSPSSLSASVGDRVTITCKASQDVYNAVAWYQQKPGQ SPKLLIYSASSRYTGVPSRFTGSGSGPDFTFTISSVQAEDLAV YFCQQHFRTPFTFGSGTKLEIK SEQIDNO:830 SLT-1A1-WT KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQIDNO:831 SLT-1A-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:832 SLT-1A-D1- KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG C242 IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVAA SEQIDNO:833 SLT-1A-D2 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSATSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASRVAR SEQIDNO:834 IA-SLT-1A-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQIDNO:835 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID protein1 SGSGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:836 cell-targeting MEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG protein2 QAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV YYCQQRSDWPLTFGGGTKVEIKGSTSGSGKPGSGEGSAVQL VESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKG LEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSSEF PKPSTPPGSSGGAPKEFTLDFSTAKTYVDSLNVIRSAIGTPLQ TISSGGTSLLMIDSGIGDNLFAVDILGFDFTLGRFNNLRLIVER NNLYVTGFVNRTNNVFYRFADFSHVTFPGTTAVTLSADSSY TTLQRVAGISRTGMQINRHSLTTSYLDLMSHSATSLTQSVAR AMLRFVTVTAEALRFRQIQRGFRTTLDDLSGASYVMTAEDV DLTLNWGRLSSVLPDYHGQDSVRVGRISFGSINAILGSVALIL NCHHHASRVAR SEQIDNO:837 cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID protein3 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASAVA AEFPKPSTPPGSSGGAPDIVLTQSPASLAVSLGQRATISCRAT ESVEYYGTSLVQWYQQKPGQPPKLLIYAASSVDSGVPARFS GSGSGTDFSLTIHPVEEDDIAMYFCQQSRRVPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKP GASVKMSCKASGYTFTSYVMHWVKQKPGQGLEWIGYVNP FNDGTKYNEMFKGKATLTSDKSSSTAYMELSSLTSEDSAVY YCARQAWGYPWGQGTLVTVSA SEQIDNO:838 referencecell- MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID targeting SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR molecule1 TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:839 referencecell- MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID targeting SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR molecule2 TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIVLTQSPASLAVSLGQRATISCRAT ESVEYYGTSLVQWYQQKPGQPPKLLIYAASSVDSGVPARFS GSGSGTDFSLTIHPVEEDDIAMYFCQQSRRVPYTFGGGTKLEI KGGGGSGGGGSGGGGSGGGGSGGGGSEVQLQQSGPELVKP GASVKMSCKASGYTFTSYVMEIWVKQKPGQGLEWIGYVNP FNDGTKYNEMFKGKATLTSDKSSSTAYMELSSLTSEDSAVY YCARQAWGYPWGQGTLVTVSA SEQIDNO:840 referencecell- MEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPG targeting QAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAV molecule3 YYCQQRSDWPLTFGGGTKVEIKGSTSGSGKPGSGEGSAVQL VESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKG LEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSSEF PKPSTPPGSSGGAPKEFTLDFSTAKTYVDSLNVIRSAIGTPLQ TISSGGTSLLMIDSGSGDNLFAVDILGFDFTLGRFNNLRLIVE RNNLYVTGFVNRTNNVFYRFADFSHVTFPGTTAVTLSADSS YTTLQRVAGISRTGMQINRHSLTTSYLDLMSHSATSLTQSVA RAMLRFVTVTAEALRFRQIQRGFRTTLDDLSGASYVMTAED VDLTLNWGRLSSVLPDYHGQDSVRVGRISFGSINAILGSVALI LNSHHHASRVAR SEQIDNO:841 referencecell- MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID targeting SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR molecule4 TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AAHHSEDPSSKAPKAPEVQLVESGGGLVQAGGSLRLSCAAS GITFSINTMGWYRQAPGKQRELVALISSIGDTYYADSVKGRF TISRDNAKNTVYLQMNSLKPEDTAVYYCKRFRTAAQGTDY WGQGTQVTVSSA SEQIDNO:842 referencecell- MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID targeting SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR molecule5 TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQIDNO:843 referencecell- MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID targeting SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR molecule1 TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQIDNO:844 heavychainABR1 YTFTSYVMH SEQIDNO:845 heavychainABR2 WIGYINPYNDGTKY SEQIDNO:846 heavychainABR3 RGTYYYGSRVFDY SEQIDNO:847 lightchainABR1 KSLLNSNGNTYLY SEQIDNO:848 lightchainABR2 LLIYRMSNLAS SEQIDNO:849 lightchainABR3 MQHLEYPF SEQIDNO:850 heavychainABR1 YAFSSYWMN SEQIDNO:851 heavychainABR2 WIGQIWPGDGDTNY SEQIDNO:852 heavychainABR3 RRETTTVGRYYYAMDY SEQIDNO:853 lightchainABR1 QSVDYDGDSYLN SEQIDNO:854 lightchainABR2 LLIYDASNLVS SEQIDNO:855 lightchainABR3 QQSTEDPW SEQIDNO:856 heavychainABR2 WIGQIWPGDGDTNYNG SEQIDNO:857 heavychainABR1 GSISTSGMGVG SEQIDNO:858 heavychainABR2 WIGHIWWDDDKRY SEQIDNO:859 heavychainABR3 RMELWSYYFDY SEQIDNO:860 lightchainABR1 SSVSYMH SEQIDNO:861 lightchainABR2 LLIYDTSKLAS SEQIDNO:862 lightchainABR3 FQGSVYPF SEQIDNO:863 heavychainCDR1 GYTFTSYNMH SEQIDNO:864 heavychainCDR2 AIYPGNGDTSYNQKFKG SEQIDNO:865 heavychainCDR3 AQLRPNYWYFDV SEQIDNO:866 lightchainCDR1 RASSSVSYMH SEQIDNO:867 lightchainCDR2 ATSNLAS SEQIDNO:868 lightchainCDR3 QQWISNPPT SEQIDNO:869 heavychainCDR1 GYTFTSYNVH SEQIDNO:870 heavychainCDR3 SNYYGSSYVWFFDV SEQIDNO:871 lightchainCDR1 RASSSVSYMD SEQIDNO:872 heavychainCDR3 STYYGGDWYFNV SEQIDNO:873 lightchainCDR1 RASSSVSYIH SEQIDNO:874 lightchainCDR3 QQWTSNPPT SEQIDNO:875 heavychainCDR1 GFTFNDYAMH SEQIDNO:876 heavychainCDR2 TISWNSGSIGYADSVKG SEQIDNO:877 heavychainCDR3 DIQYGNYYYGMDV SEQIDNO:878 lightchainCDR1 RASQSVSSYLA SEQIDNO:879 lightchainCDR2 DASNRAT SEQIDNO:880 lightchainCDR3 QQRSNWPIT SEQIDNO:881 heavychainCDR1 GYTFTSYNMH SEQIDNO:882 heavychainCDR3 VVYYSNSYWYFDV SEQIDNO:883 lightchainCDR2 APSNLAS SEQIDNO:884 lightchainCDR3 QQWSFNPPT SEQIDNO:885 heavychainCDR1 GYAFSYSWIN SEQIDNO:886 heavychainCDR2 RIFPGDGDTDYNGKFKG SEQIDNO:887 heavychainCDR3 NVFDGYWLVY SEQIDNO:888 lightchainCDR1 RSSKSLLHSNGITYLY SEQIDNO:889 lightchainCDR2 QMSNLVS SEQIDNO:890 lightchainCDR3 AQNLELPYT SEQIDNO:891 heavychainABR1 YRFTNYWIH SEQIDNO:892 heavychainABR2 WIGGINPGNNYATYRR SEQIDNO:893 heavychainABR3 TREGYGNYGAWFAY SEQIDNO:894 lightchainABR1 QSLANSYGNTFLS SEQIDNO:895 lightchainABR2 LLIYGISNRFS SEQIDNO:896 lightchainABR3 LQGTHQPY SEQIDNO:897 heavychainABR1 FAFSIYDMS SEQIDNO:898 heavychainABR2 WVAYISSGGGTTYY SEQIDNO:899 heavychainABR3 RHSGYGTHWGVLFAY SEQIDNO:900 lightchainABR1 QDISNYLA SEQIDNO:901 lightchainABR2 LLIYYTSILHS SEQIDNO:902 lightchainABR3 QQGNTLPW SEQIDNO:903 heavychainABR1 YTFTSYWLH SEQIDNO:904 heavychainABR2 WIGYINPRNDYTEY SEQIDNO:905 heavychainABR3 RRDITTFY SEQIDNO:906 lightchainABR1 QSVLYSANHKNYLA SEQIDNO:907 lightchainABR2 LLIYWASTRES SEQIDNO:908 lightchainABR3 HQYLSSW SEQIDNO:909 heavychainABR1 YEFSRSWMN SEQIDNO:910 heavychainABR2 WVGRIYPGDGDTNYSGKF SEQIDNO:911 heavychainABR3 RDGSSWDWYFDV SEQIDNO:912 lightchainABR1 QSIVHSVGNTFLE SEQIDNO:913 lightchainABR2 LLIYKVSNRFS SEQIDNO:914 lightchainABR3 FQGSQFPY SEQIDNO:915 heavychainCDR1 GYRFTNYWIH SEQIDNO:916 heavychainCDR2 GINPGNNYATYRRKFQG SEQIDNO:917 heavychainCDR3 EGYGNYGAWFAY SEQIDNO:918 lightchainCDR1 RSSQSLANSYGNTFLS SEQIDNO:919 lightchainCDR2 GISNRFS SEQIDNO:920 lightchainCDR3 LQGTHQPYT SEQIDNO:921 heavychainCDR1 GFAFSIYDMS SEQIDNO:922 heavychainCDR2 YISSGGGTTYYPDTVKG SEQIDNO:923 heavychainCDR3 HSGYGTHWGVLFAY SEQIDNO:924 lightchainCDR1 RASQDISNYLA SEQIDNO:925 lightchainCDR2 YTSILHS SEQIDNO:926 lightchainCDR3 QQGNTLPWT SEQIDNO:927 heavychainCDR1 GYTFTDYYIT SEQIDNO:928 heavychainCDR2 WIYPGSGNTKYNEKF SEQIDNO:929 heavychainCDR3 YGNYWFAY SEQIDNO:930 lightchainCDR1 KASQSVDFDGDSYMN SEQIDNO:931 lightchainCDR2 AASNLES SEQIDNO:932 lightchainCDR3 QQSNEDPWT SEQIDNO:933 heavychainCDR1 YTFTTYWMH SEQIDNO:934 heavychainCDR2 WIGYINPSTGYTDY SEQIDNO:935 heavychainCDR3 TRRGPSYGNHGAWFPY SEQIDNO:936 lightchainCDR1 ENVDTYVS SEQIDNO:937 lightchainCDR2 LLIYGASNRYT SEQIDNO:938 lightchainCDR3 GQSYRYPP SEQIDNO:939 heavychainCDR1 GYTFTGYYMH SEQIDNO:940 heavychainCDR2 WIDPNSGATTYAQKF SEQIDNO:941 heavychainCDR3 KTTQTTWGFPF SEQIDNO:942 lightchainCDR1 RASQGVYQWLA SEQIDNO:943 lightchainCDR2 KASHLYN SEQIDNO:944 lightchainCDR3 QQLNSYPLT SEQIDNO:945 heavychainCDR1 GYTFTDYWMH SEQIDNO:946 heavychainCDR2 WIGYINPNTAYTDY SEQIDNO:947 lightchainCDR1 KASENVDSFVS SEQIDNO:948 lightchainCDR2 GASNRYT SEQIDNO:949 lightchainCDR3 GQNYRYPLT SEQIDNO:950 heavychainABR1 FSLISYGVH SEQIDNO:951 heavychainABR2 WLGVIWRGGSTDY SEQIDNO:952 heavychainABR3 KTLITTGYAMDY SEQIDNO:953 lightchainABR1 EDIYNRLA SEQIDNO:954 lightchainABR2 LLISGATSLETG SEQIDNO:955 lightchainABR3 QQYWSTP SEQIDNO:956 heavychainABR1 FTFNSFAMS SEQIDNO:957 heavychainABR2 WVSAISGSGGGTYY SEQIDNO:958 heavychainABR3 KDKILWFGEPVFDY SEQIDNO:959 lightchainABR1 QSVSSYLA SEQIDNO:960 lightchainABR2 LLIYDASNRAT SEQIDNO:961 lightchainABR3 QQRSNWPP SEQIDNO:962 heavychainABR1 FSLTSYGVH SEQIDNO:963 heavychainABR2 WIGVMWRGGSTDY SEQIDNO:964 heavychainABR3 KSMITTGFVMDS SEQIDNO:965 lightchainABR1 EDIYNRLT SEQIDNO:966 lightchainABR2 LLISGATSLET SEQIDNO:967 lightchainABR3 QQYWSNPY SEQIDNO:968 heavychainABR1 FDFSRSWMN SEQIDNO:969 heavychainABR2 WIGEINPDSSTINY SEQIDNO:970 heavychainABR3 RYGNWFPY SEQIDNO:971 lightchainABR1 QNVDTNVA SEQIDNO:972 lightchainABR2 ALIYSASYRYS SEQIDNO:973 lightchainABR3 QQYDSYPL SEQIDNO:974 heavychainABR1 GTFSSYAFS SEQIDNO:975 heavychainABR2 WMGRVIPFLGIANS SEQIDNO:976 heavychainABR3 RDDIAALGPFDY SEQIDNO:977 lightchainABR1 QGISSWLA SEQIDNO:978 lightchainABR2 SLIYAASSLQS SEQIDNO:979 lightchainABR3 QQYNSYPR SEQIDNO:980 heavychainABR1 YTFTDYWMQ SEQIDNO:981 heavychainABR2 WIGTIYPGDGDTGY SEQIDNO:982 heavychainABR3 RGDYYGSNSLDY SEQIDNO:983 lightchainABR1 QDVSTVVA SEQIDNO:984 lightchainABR2 RLIYSASYRYI SEQIDNO:985 lightchainABR3 QQHYSPPY SEQIDNO:986 heavychainCDR1 GFSLTSYGVH SEQIDNO:987 heavychainCDR2 VMWRGGSTDYNAAFMS SEQIDNO:988 heavychainCDR3 SMITTGFVMDS SEQIDNO:989 lightchainCDR1 KASEDIYNRLT SEQIDNO:990 lightchainCDR2 GATSLET SEQIDNO:991 lightchainCDR3 QQYWSNPYT SEQIDNO:992 heavychainCDR1 GFSLISYGVH SEQIDNO:993 heavychainCDR2 VIWRGGSTDYNAAFMS SEQIDNO:994 heavychainCDR3 TLITTGYAMDY SEQIDNO:995 lightchainCDR1 KASEDIYNRLA SEQIDNO:996 lightchainCDR2 GATSLET SEQIDNO:997 lightchainCDR3 QQYWSTPT SEQIDNO:998 heavychainCDR1 GFDFSRSWMN SEQIDNO:999 heavychainCDR2 EINPDSSTINYTTSLKD SEQIDNO:1000 heavychainCDR3 YGNWFPY SEQIDNO:1001 lightchainCDR1 KASQNVDTNVA SEQIDNO:1002 lightchainCDR2 SASYRYS SEQIDNO:1003 lightchainCDR3 QQYDSYPLT SEQIDNO:1004 heavychainABR1 FDFSRYWMS SEQIDNO:1005 heavychainABR2 WIGEINPTSSTINF SEQIDNO:1006 heavychainABR3 RGNYYRYGDAMDY SEQIDNO:1007 lightchainABR1 KSVSTSGYSYLH SEQIDNO:1008 lightchainABR2 LLIYLASNLES SEQIDNO:1009 lightchainABR3 QHSRELPF SEQIDNO:1010 heavychainABR1 STFTTYWIH SEQIDNO:1011 heavychainABR2 WIGYINPNTGYTEY SEQIDNO:1012 heavychainABR3 VRFITVVGG SEQIDNO:1013 lightchainABR1 SSVSSSHLH SEQIDNO:1014 lightchainABR2 LWIYSTSNLAS SEQIDNO:1015 lightchainABR3 HQYHRSPL SEQIDNO:1016 heavychainABR1 FSLTTYGIGVG SEQIDNO:1017 heavychainABR2 WLTHIWWNDNKYY SEQIDNO:1018 heavychainABR3 YGYTY SEQIDNO:1019 lightchainABR1 QSLLYSNGNTYLH SEQIDNO:1020 lightchainABR2 LLIYKLSNRFS SEQIDNO:1021 lightchainABR3 SQSTHVPW SEQIDNO:1022 heavychainABR1 FNIKDTYIH SEQIDNO:1023 heavychainABR2 WVARIYPTNGYTRY SEQIDNO:1024 heavychainABR3 RWGGDGFYAMDY SEQIDNO:1025 lightchainABR1 QDVNTAVA SEQIDNO:1026 lightchainABR2 LLIYSASFLYS SEQIDNO:1027 lightchainABR3 QQHYTTPP SEQIDNO:1028 heavychainABR3 RWGGDGFYAMDV SEQIDNO:1029 heavychainABR1 YSFTSYWIA SEQIDNO:1030 heavychainABR2 YMGLIYPGDSDTKY SEQIDNO:1031 heavychainABR3 RHDVGYCSSSNCAKWPEYFQH SEQIDNO:1032 lightchainABR1 SSNIGNNYVS SEQIDNO:1033 lightchainABR2 LLIYGHTNRPA SEQIDNO:1034 lightchainABR3 AAWDDSLSGW SEQIDNO:1035 heavychainABR1 YPFTNYGMN SEQIDNO:1036 heavychainABR2 WMGWINTSTGESTF SEQIDNO:1037 heavychainABR3 RWEVYHGYVPY SEQIDNO:1038 lightchainABR1 QDVYNAVA SEQIDNO:1039 lightchainABR2 LLIYSASSRYT SEQIDNO:1040 lightchainABR3 QQHFRTPF SEQIDNO:1041 heavychainABR1 ITFSINTMG SEQIDNO:1042 heavychainABR2 LVALISSIGDTYYA SEQIDNO:1043 heavychainABR3 KRFRTAAQGTDY SEQIDNO:1044 heavychainCDR1 GFNIKDTYIH SEQIDNO:1045 heavychainCDR2 RIYPTNGYTRYADSVKG SEQIDNO:1046 heavychainCDR3 WGGDGFYAMDY SEQIDNO:1047 lightchainCDR1 RASQDVNTAVA SEQIDNO:1048 lightchainCDR2 SASFLYS SEQIDNO:1049 lightchainCDR3 QQHYTTPPT SEQIDNO:1050 heavychainCDR1 GFNIKDTYIH SEQIDNO:1051 heavychainCDR2 RIYPTNGYTRYADSVKG SEQIDNO:1052 heavychainCDR3 WGGDGFYAMDV SEQIDNO:1053 lightchainCDR1 RASQDVNTAVA SEQIDNO:1054 lightchainCDR2 SASFLYS SEQIDNO:1055 lightchainCDR3 QQHYTTPPT SEQIDNO:1056 heavychainCDR1 GYSFTSYWIA SEQIDNO:1057 heavychainCDR2 LIYPGDSDTKYSPSFQG SEQIDNO:1058 heavychainCDR3 HDVGYCSSSNCAKWPEYFQH SEQIDNO:1059 lightchainCDR1 SGSSSNIGNNYVS SEQIDNO:1060 lightchainCDR2 GHTNRPA SEQIDNO:1061 lightchainCDR3 AAWDDSLSGWV SEQIDNO:1062 heavychainCDR1 GITFSINTMG SEQIDNO:1063 heavychainCDR2 LISSIGDTYYADSVKG SEQIDNO:1064 heavychainCDR3 FRTAAQGTDY SEQIDNO:1065 heavychainABR1 FTFSDSWIH SEQIDNO:1066 heavychainABR2 WVAWISPYGGSTYY SEQIDNO:1067 heavychainABR3 RRHWPGGFDY SEQIDNO:1068 lightchainABR1 QDVSTAVA SEQIDNO:1069 lightchainABR2 LLIYSASFLYS SEQIDNO:1070 lightchainABR3 QQYLYHPA SEQIDNO:1071 heavychainABR1 YTFTSYVMH SEQIDNO:1072 heavychainABR2 WIGYVNPFNDGTKY SEQIDNO:1073 heavychainABR3 RQAWGYPGFQH SEQIDNO:1074 lightchainABR1 ESVEYYGTSLVQ SEQIDNO:1075 lightchainABR2 LLIYAASSVDS SEQIDNO:1076 lightchainABR3 QQSRRVPY SEQIDNO:1077 heavychainABR1 YTFTSYDVH SEQIDNO:1078 heavychainABR2 WMGWLHADTGITKF SEQIDNO:1079 heavychainABR3 RERIQLWFDY SEQIDNO:1080 lightchainABR1 QGISSWLA SEQIDNO:1081 lightchainABR2 SLIYAASSLQS SEQIDNO:1082 lightchainABR3 QQYNSYPY SEQIDNO:1083 heavychainABR1 DTFSTYAIS SEQIDNO:1084 heavychainABR2 WMGGIIPIFGKAHY SEQIDNO:1085 heavychainABR3 RKFHFVSGSPFGMDV SEQIDNO:1086 lightchainABR1 QSVSSYLA SEQIDNO:1087 lightchainABR2 LLIYDASNRAT SEQIDNO:1088 lightchainABR3 QQRSNWP SEQIDNO:1089 heavychainABR1 FTFSSYIMM SEQIDNO:1090 heavychainABR2 WVSSIYPSGGITFY SEQIDNO:1091 heavychainABR3 RIKLGTVTTVDY SEQIDNO:1092 lightchainABR1 SSDVGGYNYVS SEQIDNO:1093 lightchainABR2 LMIYDVSNRPS SEQIDNO:1094 lightchainABR3 SSYTSSSTR SEQIDNO:1095 heavychainCDR1 GFNIKDYFLH SEQIDNO:1096 heavychainCDR2 WINPDNGNTVYDKFQG SEQIDNO:1097 heavychainCDR3 RDYTYEKAALDY SEQIDNO:1098 lightchainCDR1 RASGNIYNYLA SEQIDNO:1099 lightchainCDR2 DAKTLAD SEQIDNO:1100 lightchainCDR3 QHFWSLPFT SEQID SLT-1A-Cys5-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMIDSG NO:1101 (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQID SLT-1A-Cys7-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1102 (inactivated) CGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQID SLT-1A-Cys8-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1103 (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHCGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTLDDLSGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQID SLT-1A-Cys9-D1 KEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1104 (inactivated) IGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNRTN variant2 NVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTADAL RFRQIQRGFRTTLDDLCGASYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVAA SEQID cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID NO:1105 protein4 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQID cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID NO:1106 protein5 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS SEQID cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID NO:1107 protein6 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAE ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQID cell-targeting MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMID NO:1108 protein7 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSGGGGSGGGGSGGGGSGGGGSQVQLQESGPGLVRPSQTLS LTCTVSGFSLTSYGVHWVRQPPGRGLEWIGVMWRGGSTDY NAAFMSRLNITKDNSKNQVSLRLSSVTAADTAVYYCAKSMI TTGFVMDSWGQGSLVTVSS SEQID StxA-Lys(1) KEFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1109 TGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR MASDEFPSMCPADGRVRGITHNAILWDSSTLGAILMRRTISS SEQID StxA-Lys(11) AEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1110 TGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR MASDEFPSMCPADGRVRGITHNAILWDSSTLGAILMRRTISS SEQID StxA-Lys(274) AEFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1111 TGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR MASDEFPSMCPADGRVRGITHNKILWDSSTLGAILMRRTISS SEQID StxA1-Lys(1) KEFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1112 TGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRT NNVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTG MQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEA LRFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSV LPDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQID StxA-Lys(11) KTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGTGDNLFAVD NO:1113 withamino- VRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTNNVFYRFADF terminal SHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGMQINRHSLTT truncation SYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRFRQIQRGF RTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVLPDYHGQDS VRVGRISFGSINAILGSVALILNCHHHASRVARMASDEFPSM CPADGRVRGITHNAILWDSSTLGAILMRRTISS SEQID StxA1-Lys(11) KTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGTGDNLFAVD NO:1114 withamino- VRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTNNVFYRFADF terminal SHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGMQINRHSLTT truncation SYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRFRQIQRGF RTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVLPDYHGQDS VRVGRISFGSINAILGSVALILNCHHHASRVAR SEQID StxA-Lys(274) EFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGT NO:1115 withamino- GDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN terminal NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM truncationvariant QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL 1 RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVARM ASDEFPSMCPADGRVRGITHNKILWDSSTLGAILMRRTISS SEQID StxA1-Lys(274) TYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGTGDNLFAVDV NO:1116 withamino- RGIDPEEGRFNNLRLIVERNNLYVTGFVNRTNNVFYRFADFS terminal HVTFPGTTAVTLSGDSSYTTLQRVAGISRTGMQINRHSLTTS truncationvariant YLDLMSHSGTSLTQSVARAMLRFVTVTAEALRFRQIQRGFR 2 TTLDDLSGRSYVMTAEDVDLTLNWGRLSSVLPDYHGQDSV RVGRISFGSINAILGSVALILNCHHHASRVARMASDEFPSMCP ADGRVRGITHNKILWDSSTLGAILMRRTISS SEQID SLT-1A-Lys(1) KEFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1117 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVARM ASDEFPSMCPADGRVRGITHNAILWDSSTLGAILMRRTISS SEQID SLT-1A-Lys(11) AEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1118 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVARM ASDEFPSMCPADGRVRGITHNAILWDSSTLGAILMRRTISS SEQID ST-1A-Lys(274) AEFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1119 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVARM ASDEFPSMCPADGRVRGITHNKILWDSSTLGAILMRRTISS SEQID SLT-1A1-Lys(1) KEFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSG NO:1120 SGDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVAR SEQID SLT-1A-Lys(11) KTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGSGDNLFAVD NO:1121 withamino- VRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTNNVFYRFADF terminal SHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGMQINRHSLTT truncationvariant SYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRFRQIQRGF 1 RTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVLPDYHGQDS VRVGRISFGSINAILGSVALILNCHHHASRVARMASDEFPSM CPADGRVRGITHNAILWDSSTLGAILMRRTISS SEQID SLT-1A1-Lys(11) KTYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGSGDNLFAVD NO:1122 withamino- VRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTNNVFYRFADF terminal SHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGMQINRHSLTT truncationvariant SYLDLMSHSGTSLTQSVARAMLRFVTVTAEALRFRQIQRGF 2 RTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVLPDYHGQDS VRVGRISFGSINAILGSVALILNCHHHASRVAR SEQID SLT-1A-Lys(274) EFTLDFSTAATYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGS NO:1123 withamino- GDNLFAVDVRGIDPEEGRFNNLRLIVERNNLYVTGFVNRTN terminal NVFYRFADFSHVTFPGTTAVTLSGDSSYTTLQRVAGISRTGM truncationvariant QINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAEAL 1 RFRQIQRGFRTTLDDLSGRSYVMTAEDVDLTLNWGRLSSVL PDYHGQDSVRVGRISFGSINAILGSVALILNCHHHASRVARM ASDEFPSMCPADGRVRGITHNKILWDSSTLGAILMRRTISS SEQID SLT-1A-Lys(274) TYVDSLNVIRSAIGTPLQTISSGGTSLLMIDSGSGDNLFAVDV NO:1124 withamino- RGIDPEEGRFNNLRLIVERNNLYVTGFVNRTNNVFYRFADFS terminal HVTFPGTTAVTLSGDSSYTTLQRVAGISRTGMQINRHSLTTS truncationvariant YLDLMSHSGTSLTQSVARAMLRFVTVTAEALRFRQIQRGFR 2 TTLDDLSGRSYVMTAEDVDLTLNWGRLSSVLPDYHGQDSV RVGRISFGSINAILGSVALILNCHHHASRVARMASDEFPSMCP ADGRVRGITHNKILWDSSTLGAILMRRTISS SEQID SLT-2A-Lys(11) DEFTVDFSSQKSYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1125 VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ AQATECQIVGDRAAIAVNNVLWEANTIAALLNRAPQDLTEP NQ SEQID SLT-2A-Lys(255) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1126 VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ KQATECQIVGDRAAIAVNNVLWEANTIAALLNRAPQDLTEP NQ SEQID SLT-2A-Lys(257) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1127 VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ AQKTECQIVGDRAAIAVNNVLWEANTIAALLNRAPQDLTEP NQ SEQID SLT-2A-Lys(270) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1128 VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ AQATECQIVGDRAAIKVNNVLWEANTIAALLNRAPQDLTEP NQ SEQID SLT-2A-Lys(288) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1129 VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ AQATECQIVGDRAAIAVNNVLWEANTIAALLNRKPQDLTEP NQ SEQID SLT-2A1-Lys(11) DEFTVDFSSQKSYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1130 VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVR SEQID SLT-2A-Lys(255) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1131 withamino- RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncation DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQKQATECQIVGDR AAIAVNNVLWEANTIAALLNRAPQDLTEPNQ SEQID SLT-2A-Lys(257) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1132 withamino- RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncation DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQAQKTECQIVGDR AAIAVNNVLWEANTIAALLNRAPQDLTEPNQ SEQID SLT-2A-Lys(270) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1133 withamino- RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncation DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQAQATECQIVGDR AAIKVNNVLWEANTIAALLNRAPQDLTEPNQ SEQID SLT-2A-Lys(288) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1134 withamino- RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncation DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQAQATECQIVGDR AAIAVNNVLWEANTIAALLNRKPQDLTEPNQ SEQID SLT-2A-Lys(255) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1135 withcarboxy- VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE terminal TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM truncation QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ K SEQID SLT-2A-Lys(257) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1136 withcarboxy- VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE terminal TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM truncation QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ AQK SEQID SLT-2A-Lys(270) DEFTVDFSSQASYVDSLNSIRSAISTPLGNISQGGVSVSVINH NO:1137 withcarboxy- VLGGNYISLNVRGLDPYSERFNHLRLIMERNNLYVAGFINTE terminal TNIFYRFSDFSHISVPDVITVSMTTDSSYSSLQRIADLERTGM truncation QIGRHSLVGSYLDLMEFRGRSMTRASSRAMLRFVTVIAEAL RFRQIQRGFRPALSEASPLYTMTAQDVDLTLNWGRISNVLPE YRGEEGVRIGRISFNSLSAILGSVAVILNCHSTGSYSVRSVSQ AQATECQIVGDRAAIK SEQID SLT-2A-Lys(255) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1138 withamino-and RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH carboxy-terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncations DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQK SEQID SLT-2A-Lys(257) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1139 withamino-and RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH carboxy-terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncations DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQAQK SEQID SLT-2A-Lys(270) SYVDSLNSIRSAISTPLGNISQGGVSVSVINHVLGGNYISLNV NO:1140 withamino-and RGLDPYSERFNHLRLIMERNNLYVAGFINTETNIFYRFSDFSH carboxy-terminal ISVPDVITVSMTTDSSYSSLQRIADLERTGMQIGRHSLVGSYL truncations DLMEFRGRSMTRASSRAMLRFVTVIAEALRFRQIQRGFRPAL SEASPLYTMTAQDVDLTLNWGRISNVLPEYRGEEGVRIGRIS FNSLSAILGSVAVILNCHSTGSYSVRSVSQAQATECQIVGDR AAIK SEQID IA-SLT-1A- MKEFTLDFSTAKTYVDSLNVIRSAIGTPLQTISCGGTSLLMID NO:1141 Cys5-D1::scFv2 SGIGDNLFAVDILGFDFTLGRFNNLRLIVERNNLYVTGFVNR TNNVFYRFADFSHVTFPGTTAVTLSADSSYTTLQRVAGISRT GMQINRHSLTTSYLDLMSHSGTSLTQSVARAMLRFVTVTAD ALRFRQIQRGFRTTLDDLSGASYVMTAEDVDLTLNWGRLSS VLPDYHGQDSVRVGRISFGSINAILGSVALILNSHHHASAVA AEFPKPSTPPGSSGGAPDIQMTQSPSSLSASVGDRVTITCKAS EDIYNRLTWYQQKPGKAPKLLISGATSLETGVPSRFSGSGSG TDFTFTISSLQPEDIATYYCQQYWSNPYTFGQGTKVEIKGGG GSQVQLQESGPGLVRPSQTLSLTCTVSGFSLTSYGVHWVRQ PPGRGLEWIGVMWRGGSTDYNAAFMSRLNITKDNSKNQVS LRLSSVTAADTAVYYCAKSMITTGFVMDSWGQGSLVTVSS