Human immunodeficiency virus GP120 binding proteins
12583911 ยท 2026-03-24
Assignee
Inventors
- Eric ARNOULT (Branford, CT, US)
- Tiancen HU (Branford, CT, US)
- Mark R. KRYSTAL (Branford, CT, US)
- James SCHAWALDER (Durham, NC, US)
Cpc classification
C07K16/1145
CHEMISTRY; METALLURGY
C07K2317/14
CHEMISTRY; METALLURGY
International classification
Abstract
Antigen binding proteins of the invention bind to Human Immunodeficiency Virus (HIV) envelope protein and are useful in treating and preventing HIV infection. In particular, the antigen binding proteins bind to two different epitopes on HIV envelope surface glycoprotein 120 (gp120): the V3 loop region and the CD4 binding site.
Claims
1. A nucleic acid sequence that encodes an anti-HIV gp120-binding protein having two identical heavy chains and two identical light chains, comprising or consisting of: a heavy chain that is at least 95% identical to SEQ ID NO: 121 and a light chain that is at least 95% identical to SEQ ID NO:63, wherein the two heavy chains comprise a CDRH1 of SEQ ID NO:22, a CDRH2 of SEQ ID NO: 23, a CDRH3 of SEQ ID NO:24, and the two light chains comprise a CDRL1 of SEQ ID NO:25, a CDRL2 of SEQ ID NO:26 and a CDRL3 of SEQ ID NO:27.
2. An expression vector that comprises the nucleic acid sequence of claim 1.
3. A host cell that comprises the nucleic acid sequence of claim 1.
4. A host cell that comprises two expression vectors: a first expression vector comprising a first nucleic acid sequence encoding a heavy chain of SEQ ID NO: 121; and a second expression vector comprising a second nucleic acid sequence encoding a light chain of SEQ ID NO:63.
5. A method of producing an anti-HIV gp120-binding protein, comprising culturing the host cell as defined in claim 3 under conditions suitable for expression of said nucleic acid sequence, whereby the anti-HIV gp120-binding protein is produced.
6. The host cell of claim 3, wherein the host cell is a mammalian host cell.
7. The host cell of claim 6, wherein the mammalian host cell is a Chinese Hamster Ovary (CHO) cell, an NS0 cell, a PER.C6 cell, a HEK293 cell, or a HeLa cell.
8. A method of producing an anti-HIV gp120-binding protein, comprising culturing the host cell as defined in claim 4 under conditions suitable for expression of said expression vectors whereby the anti-HIV gp120-binding protein is produced.
9. The host cell of claim 4, wherein the host cell is a mammalian host cell.
10. The host cell of claim 9, wherein the mammalian host cell is a Chinese Hamster Ovary (CHO) cell, an NS0 cell, a PER.C6 cell, a HEK293 cell, or a HeLa cell.
Description
DESCRIPTION OF DRAWINGS/FIGURES
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DETAILED DESCRIPTION OF THE INVENTION
Definitions
(8) Affinity, also referred to as binding affinity, is the strength of binding at a single interaction site, i.e., of one molecule, e.g., an antigen binding protein, to another molecule, e.g., its target antigen, at a single binding site. The binding affinity of an antigen binding protein to its target may be determined by equilibrium methods (e.g., enzyme-linked immunoabsorbent assay (ELISA) or radioimmunoassay (RIA)), or kinetics (e.g., BIACORE analysis).
(9) Alternative antibody formats include alternative scaffolds in which one or more CDRs of the antigen binding protein can be arranged onto a suitable non-immunoglobulin protein scaffold or skeleton, such as an affibody, a SpA scaffold, an LDL receptor class A domain, an avimer (see, e.g., U.S. Patent Application Publication Nos. 2005/0053973, 2005/0089932, 2005/0164301) or an EGF domain.
(10) Antibody is used herein to refer to a heterotetrameric glycoprotein with an approximate molecular weight of 150,000 daltons. An intact antibody is composed of two identical heavy chains (HCs) and two identical light chains (LCs) linked by covalent disulphide bonds. This H2L2 structure folds to form a Y shape with three functional domains comprising two antigen-binding fragments, known as Fab fragments (the top of the Y), and a fragment crystallisable Fc (the bottom of the Y). The Fab fragment is composed of the variable domain at the amino-terminus, variable heavy (VH) or variable light (VL), and the constant domain at the carboxyl terminus, CH1 (heavy) and CL (light). The Fc fragment is composed of two domains formed by dimerization of paired CH2 and CH3 regions. The Fc may elicit effector functions by binding to receptors on immune cells or by binding Clq, the first component of the classical complement pathway. The five classes of antibodies IgM, IgA, IgG, IgE and IgD are defined by distinct heavy chain amino acid sequences, which are called , , , and respectively; each heavy chain can pair with either a K or A light chain. The majority of antibodies in the serum belong to the IgG class, there are four isotypes of human IgG (IgG1, IgG2, IgG3 and IgG4), the sequences of which differ mainly in their hinge region. In an embodiment, an anti-CD4bs antibody, as used herein, refers to an antibody that binds to a CD4 binding site
(11) Antigen binding antibody fragments or antigen binding fragments or antibody fragments as used herein include Fab, F(ab).sub.2, Fv, disulphide linked Fv, single chain Fv (scFv), disulphide-linked scFv, diabodies, TANDABS, etc. and modified versions of any of the foregoing (for a summary of alternative antibody formats see Holliger and Hudson, Nature Biotechnology, 23 (9), 1126-1136, 2005).
(12) Antigen binding protein and anti-gp120 binding protein are used interchangeably herein and refer to antibodies and fragments thereof, alternative antibody formats, and other protein constructs, such as domains, that are capable of binding to HIV gp120. Envelope glycoprotein gp120 (or gp120) is a 120 kDa glycoprotein that is part of the outer layer of HIV. It presents itself as viral membrane spikes consisting of three molecules of gp120 linked together and anchored to the membrane by gp41 protein. Gp120 is essential for viral infection as it facilitates HIV entry into the host cell through its interaction with cell surface receptors. Gp120 is encoded by the HIV env gene. The env gene encodes a gene product of around 850 amino acids. The primary env product is the protein gp160, which gets cleaved into gp120 (about 480 amino acids) and gp41 (about 345 amino acids) in the endoplasmic reticulum by the cellular protease furin. The amino acid sequence of an exemplary gp160 from HIV clone WITO is provided below (SEQ ID NO: 363; the V3 loop is boldened and the potential N332 N-linked glycosylation site is boldened and underlined):
(13) TABLE-US-00001 MKVMGTKKNYQHLWRWGIMLLGMLMMSSAAEQLWVTVYYGVPVWR EANTTLFCASDAKAYDTEVHNVWATHACVPTDPNPQEVVMGNVTE DFNMWKNNMVEQMHEDIISLWDQSLKPCVKLTPLCVTLHCTNVTI SSTNGSTANVTMREEMKNCSFNTTTVIRDKIQKEYALFYKLDIVP IEGKNTNTSYRLINCNTSVITQACPKVSFEPIPIHYCAPAGFAIL KCNNKTFNGKGPCRNVSTVQCTHGIKPVVSTQLLLNGSLAEEDII IRSENFTNNGKNIIVQLKEPVKINCTRPGNNTRRSINIGPGRAFY ATGAIIGDIRKAHCNISTEQWNNTLTQIVDKLREQFGNKTIIFNQ SSGGDPEVVMHTFNCGGEFFYCNSTQLFNSTWENNGTSTWNSTAD NITLPCRIKQVINMWQEVGKAMYAPPIRGQIDCSSNITGLILTRD GGSNSSQNETFRPGGGNMKDNWRSELYKYKVVKIEPLGIAPTRAK RRVVQREKRAVTLGAVFLGFLGAAGSTMGAASLTLTVQARLLLSG IVQQQSNLLRAIEAQQHMLQLTVWGIKQLQARVLAIERYLKDQQL LGIWGCSGKLICTTTVPWNTSWSNKSYDYIWNNMTWMQWEREIDN YTGFIYTLIEESQNQQEKNELELLELDKWASLWNWFNITNWLWYI KLFIMIIGGLVGLRIVCAVLSIVNRVRQGYSPLSFQTRLPNPRGP DRPEETEGEGGERDRDRSARLVNGFLAIIWDDLRSLCLFSYHRLR DLLLIVARVVEILGRRGWEILKYWWNLLKYWSQELKNSAVSLLNV TAIAVAEGTDRVIEIVQRAVRAILHIPTRIRQGFERALL
The amino acid of an exemplary gp120 is provided below (SEQ ID NO: 364; the V3 loop is boldened and the potential N332 N-linked glycosylation site is boldened and underlined):
(14) TABLE-US-00002 AEQLWVIVYYGVPVWREANTTLFCASDAKAYDTEVHNVWATHACV PTDPNPQEVVMGNVTEDFNMWKNNMVEQMHEDIISLWDQSLKPCV KLTPLCVTLHCTNVTISSTNGSTANVTMREEMKNCSFNTTTVIRD KIQKEYALFYKLDIVPIEGKNTNTSYRLINCNTSVITQACPKVSF EPIPIHYCAPAGFAILKCNNKTFNGKGPCRNVSTVQCTHGIKPVV STQLLLNGSLAEEDIIIRSENFTNNGKNIIVQLKEPVKINCTRPG NNTRRSINIGPGRAFYATGAIIGDIRKAHCNISTEQWNNTLTQIV DKLREQFGNKTIIFNQSSGGDPEVVMHTFNCGGEFFYCNSTQLFN STWENNGTSTWNSTADNITLPCRIKQVINMWQEVGKAMYAPPIRG QIDCSSNITGLILTRDGGSNSSQNETFRPGGGNMKDNWRSELYKY KVVKIEPLGIAPTRAKRRVVQREKR
(15) Antigen binding site and paratope are used interchangeably herein and refer to a particular site on an antigen binding protein that makes contact with and is capable of specifically binding to a site (i.e., epitope) on an antigen, e.g., HIV gp120. The antigen binding site may be formed by a single variable domain, or paired VH/VL domains as can be found on a standard antibody. Single-chain Fv (ScFv) domains can also provide antigen binding sites.
(16) Avidity also referred to as functional affinity, is the cumulative strength of binding at multiple interaction sites, e.g., the sum total of the strength of binding of two molecules (or more) to one another at multiple sites, e.g., taking into account the valency of the interaction.
(17) A bispecific molecule as used herein is an antigen binding protein that is capable of binding to two different epitopes on the same antigen, i.e., HIV gp120 protein. In particular, one epitope comprises part of or the whole of the V3 loop region of gp120 and the other epitope comprises part of or the whole of the CD4 binding site of gp120.
(18) Broadly neutralizing antibody or bNAb as used herein, is meant an antibody that neutralizes more than one HIV-1 virus species (from diverse clades and different strains within a clade) in a neutralization assay. A broad neutralizing antibody may neutralize at least 2, 3, 4, 5, 6, 7, 8, 9 or more different strains of HIV-1, the strains belonging to the same or different clades.
(19) CD4 binding site or CD4-binding site or CD4bs refers to a site on the HIV envelope protein gp120 that binds to CD4. (Cluster of differentiation factor 4). CD4 is a T-cell surface protein that serves as the primary receptor site for HIV during HIV infection. The CD4 binding site on gp120 is a highly conserved, discontinuous and conformational that comprises residues on either side of the HIV V4 loop (Curr HIV/AIDS Rep, 9 (1): 52-63, 2021) that binds to CD4.
(20) A CD4 domain as used herein is a soluble recombinant form of human CD4 (Cluster of differentiation factor 4, a transmembrane glycoprotein found on T-cells), or a fragment thereof, that mimics the activity of native membrane-anchored human CD4 in its binding interactions with the HIV envelope protein. A CD4 domain of the present invention binds to the CD4-binding site of HIV gp120 and may block the ability of HIV gp120 to bind membrane-anchored CD4, e.g., on CD4+ T cells. A CD4 domain of the invention may induce a structural rearrangement in gp120 upon binding, including a structural rearrangement of part or all of the V3 region of gp120. This structural rearrangement in gp120 results in a high affinity binding site for a chemokine coreceptor (CXCR4 and/or CCR5) being exposed. Native CD4 comprises four domains that are exposed on the extracellular surface of the cell, D1, D2, D3 and D4; a transmembrane domain; and a cytoplasmic tail domain. D1 and D3 resemble Ig variable domains and D2 and D4 resemble Ig constant domains. CD4 domains of the invention include one or more of domains D1 to D4 of CD4, or variants thereof. Examples of CD4 domains of the invention include wild-type D1 (SEQ ID NO:3); mD1.22 (SEQ ID NO:4), which is a variant of D1 of CD4 (Chen et al, JVI 88 (2): 1125-39, 2014); wild-type D1D2 (SEQ ID NO:1); mD1.22-D2 (SEQ ID NO: 2), which is a variant of D1D2 (Fetzer et al., Journal of Virology, 92 (12), 2018); and further variants of mD1.22 (SEQ ID NOs: 5-21).
(21) CDRs are defined as the complementarity determining region amino acid sequences of an antigen binding protein. These are the hypervariable regions of immunoglobulin heavy and light chains. There are three heavy chain and three light chain CDRs (or CDR regions) in the variable portion of an immunoglobulin. In one embodiment, the CDRs are defined based on the Kabat definition. In another embodiment, the CDRs are defined based on the Chothia definition. In a further embodiment, the Chothia definition is from Discovery Studio which uses the definitions from Chothia and Lesk, J Mol Biol. 196 (4): 901-17 (1987) and Morea et al, Methods, 20:267-279 (2000). In another embodiment, the Chothia definition is based on the Chothia from Abysis definition. In a further embodiment, the CDRs are defined based on the IMGT definition. In another embodiment, the CDRs are defined based on the Honegger definition. In another embodiment, the CDRs are defined based on the contact definition. Thus, CDRs as used herein refers to all three heavy chain CDRs, all three light chain CDRs, all heavy and light chain CDRs, or at least two CDRs.
(22) Domain refers to a folded polypeptide structure that retains its tertiary structure independent of the rest of the polypeptide. Generally, domains are responsible for discrete functional properties of polypeptides and in many cases may be added, removed or transferred to other polypeptides without loss of function of the remainder of the protein and/or of the domain.
(23) Effector Function as used herein refers to one or more of antibody-mediated effects including antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-mediated complement activation including complement-dependent cytotoxicity (CDC), complement-dependent cell-mediated phagocytosis (CDCP), antibody dependent complement-mediated cell lysis (ADCML), and Fc-mediated phagocytosis or antibody-dependent cellular phagocytosis (ADCP).
(24) Epitope as used herein refers to the portion of an antigen (e.g., gp120) that makes contact with and is capable of specifically binding to a particular site (paratope) on an antigen binding protein. An epitope may be linear or conformational/discontinuous. A conformational/discontinuous epitope comprises amino acid residues that are separated by other sequences, i.e., it does not comprise a continuous sequence in the antigen's primary amino acid sequence, but instead relies on the tertiary folding of the polypeptide. Although the residues within a confirmational/discontinuous epitope may be from different regions of the polypeptide chain, they are in close proximity in the three-dimensional structure of the antigen.
(25) In the case of multimeric antigens, a conformational or discontinuous epitope may include residues from different polypeptide chains. Particular residues comprised within an epitope can be determined through computer modelling programs or via three-dimensional structures obtained through methods known in the art, such as X-ray crystallography.
(26) Epitope mapping can be carried out using various techniques known to persons skilled in the art as described in publications such as Methods in Molecular Biology Epitope Mapping Protocols, by Mike Schutkowski and Ulrich Reineke (volume 524, 2009) and Johan Rockberg and Johan Nilvebrant (volume 1785, 2018). Exemplary methods include peptide-based approaches such as pepscan whereby a series of overlapping peptides are screened for binding using techniques such as ELISA or by in vitro display of large libraries of peptides or protein mutants, e.g., on phage. Detailed epitope information can be determined by structural techniques including X-ray crystallography, solution nuclear magnetic resonance (NMR) spectroscopy and cryogenic-electron microscopy (cryo-EM). Mutagenesis, such as alanine scanning, is an effective approach whereby loss of binding analysis is used for epitope mapping. Another method is hydrogen/deuterium exchange (HDX) combined with proteolysis and liquid-chromatography mass spectrometry (LC-MS) analysis to characterize discontinuous or conformational epitopes.
(27) Half-life or t1/2 refers to the time required for the serum concentration of an antigen binding protein to reach half of its original value. The serum half-life of proteins can be measured by pharmacokinetic studies according to the method described by Kim et al., 1994, Eur. J. of Immuno. 24:542-548. According to this method, radio-labelled protein is injected intravenously into mice and its plasma concentration is periodically measured as a function of time, for example, at about 3 minutes to about 72 hours after the injection. Other methods for pharmacokinetic analysis and determination of the half-life of a molecule will be familiar to those skilled in the art.
(28) HIV envelope protein or ENV refers to a trimeric viral membrane-associated glycoprotein (gp) or spike. It is found on both the viral membrane and the cell membrane of infected host cells. The env gene encodes the gp160 polypeptide which forms a homotrimer and is cleaved into gp120 and gp41 polypeptides. Gp120 is a surface (SU) glycoprotein responsible for binding to receptor molecules and the transmembrane (TM) glycoprotein, gp41, mediates fusion of the viral membrane with the plasma cell membrane. Over half of the mass of the trimeric envelope spike is an N-linked glycan shield that hides most amino acid-based epitopes on gp120. Binding of the cell surface receptor CD4 to HIV gp120 induces a structural rearrangement creating a high affinity binding site for a chemokine coreceptor (CXCR4 and/or CCR5), on gp120. Following gp120 binding to CXCR4 or CCR5 further conformational changes are triggered which results in gp120 disengaging from gp41, allowing for the fusion peptide of gp41 to be inserted into the cell membrane, which in turn triggers a sequence of structural changes resulting in membrane fusion (Dimitrov et al., Biochemistry 44 (37): 12471-12479, 2005).
(29) Human immunodeficiency virus (HIV) has been characterized into two types: HIV-1 and HIV-2. HIV-1 is more virulent and more infective than HIV-2 and is the cause of the majority of HIV infections globally, whereas HIV-2 is limited to a much smaller number of people, mostly in West Africa (Gilbert et al., Statistics in Medicine 22 (4): 573-593). Herein, when reference is made to HIV this is intended to mean HIV-1. HIV virions are spherical with viral glycoprotein spikes, the HIV envelope protein, protruding outwards. A conical capsid exists within the virion, enclosing a ribonucleoprotein complex comprising two copies of positive-sense single stranded RNA tightly bound to nucleocapsid proteins and enzymes needed for viral replication.
(30) A linker is an amino acid sequence that links one domain in a polypeptide to another domain in a polypeptide. For example, a linker within the meaning of the invention includes an amino acid sequence that joins a CD4 domain to a bNAb heavy chain or a bNAb light chain. In an embodiment, the linker is not cleavable under intracellular conditions.
(31) Multi-specific antigen binding protein or MSABP refers to an antigen binding protein that comprises at least two different antigen binding sites. Each of these antigen-binding sites is capable of binding to a different epitope, which may be present on the same antigen or different antigens. In an embodiment, the multi-specific antigen binding proteins of the invention are bispecific molecules capable of binding to two different epitopes on the HIV envelope protein. In particular, one epitope may comprise part of or the whole of the V3 loop region of gp120 and the other epitope may comprise part of or the whole of the CD4 binding site of gp120.
(32) Symmetric formats of MSABPs combine multiple binding specificities in a single polypeptide chain or single HL pair including Fc-fusion proteins of fragment-based formats and formats whereby antibody fragments are fused to regular antibody molecules. Examples of symmetric formats may include DVD-Ig, TVD-Ig, CODV-Ig, (scFv).sub.4-Fc, IgG-(scFv).sub.2, Tetravalent DART-Fc, F(ab).sub.4CrossMab, IgG-HC-scFv, IgG-LC-scFv, mAb-dAb etc.
(33) Neutralizes as used throughout the present specification means that the biological activity of HIV is reduced in the presence of an antigen binding protein as described herein in comparison to the biological activity of HIV in the absence of the antigen binding protein, in vitro or in vivo. For example, a neutralizing antigen binding protein of the invention may inhibit HIV entry into a target cell and reduce viral load in a patient infected with HIV.
(34) Percent identity or % identity between a query amino acid sequence and a subject amino acid sequence is the Identities value, expressed as a percentage, that is calculated using a suitable algorithm (e.g., BLASTP, FASTA, Needleman-Wunsch, Smith-Waterman, LALIGN, or GenePAST/KERR) or software (e.g., DNASTAR Lasergene, GenomeQuest, EMBOSS needle or EMBOSS infoalign), over the entire length of the query sequence after a pair-wise global sequence alignment has been performed using a suitable algorithm (e.g., Needleman-Wunsch or GenePAST/KERR) or software (e.g. DNASTAR Lasergene or GenePAST/KERR). Importantly, a query amino acid sequence may be described by an amino acid sequence disclosed herein, in particular in one or more of the claims.
(35) The query sequence may be 100% identical to the subject sequence, or it may include up to a certain integer number of amino acid alterations as compared to the subject sequence such that the % identity is less than 100%. For example, the query sequence is at least 50, 60, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical to the subject sequence. In the case of amino acid sequences, such alterations include at least one amino acid residue deletion, substitution (including conservative and non-conservative substitutions), or insertion, wherein said alterations may occur at the amino- or carboxy-terminal positions of the query sequence or anywhere between those terminal positions, interspersed either individually among the amino acid residues in the query sequence or in one or more contiguous groups within the query sequence.
(36) For antibody sequences, the % identity may be determined across the entire length of the query sequence, including the CDRs. Alternatively, the % identity may exclude one or more or all of the CDRs, for example all of the CDRs are 100% identical to the subject sequence and the % identity variation is in the remaining portion of the query sequence, e.g., the framework sequence, so that the CDR sequences are fixed and intact.
(37) Protein scaffold as used herein includes, but is not limited to, an immunoglobulin (Ig) scaffold, for example an IgG scaffold, which may be a four chain or two chain antibody, or which may comprise only the Fc region of an antibody, or which may comprise one or more constant regions from an antibody, which constant regions may be of human origin.
(38) The protein scaffold may be an Ig scaffold, for example an IgG, or IgA scaffold. The IgG scaffold may comprise some or all the domains of an intact antibody (i.e., CH1, CH2, CH3, VH, VL). The antigen binding protein may comprise an IgG scaffold selected from IgG1, IgG2, IgG3, IgG4 or IgG4PE. For example, the scaffold may be IgG1. The scaffold may consist of, or comprise, the Fc region of an antibody, or is a part thereof.
(39) The protein scaffold may be a non-Ig scaffold. The protein scaffold may be a derivative of a scaffold selected from the group consisting of CTLA-4, lipocalin, Protein A derived molecules such as Z-domain of Protein A (Affibody, SpA), A-domain (Avimer/Maxibody); heat shock proteins such as GroEI and GroES; transferrin (trans-body); ankyrin repeat protein (DARPin); peptide aptamer; C-type lectin domain (Tetranectin); human -crystallin and human ubiquitin (affilins); PDZ domains; scorpion toxin kunitz type domains of human protease inhibitors; and fibronectin/adnectin; which has been subjected to protein engineering in order to obtain binding to an antigen, such as gp120.
(40) Single variable domain refers to a folded polypeptide domain comprising sequences characteristic of antibody variable domains. It therefore includes complete antibody variable domains such as VH, VHH and VL and modified antibody variable domains, for example, in which one or more loops have been replaced by sequences that are not characteristic of antibody variable domains, or antibody variable domains that have been truncated or comprise N- or C-terminal extensions, as well as folded fragments of variable domains that retain at least the binding activity and specificity of the full-length domain. A single variable domain as defined herein is capable of binding an antigen or epitope independently of a different variable region or domain. A domain antibody or DAB may be considered the same as a human single variable domain. A single variable domain may be a human single variable domain, but also includes single variable domains from other species such as rodent (for example, as disclosed in WO 00/29004), nurse shark and Camelid VHHs Camelid VHHs are immunoglobulin single variable domain polypeptides that are derived from species including camel, llama, alpaca, dromedary, and guanaco, which produce heavy chain only antibodies naturally devoid of light chains. Such VHH domains may be humanised according to standard techniques available in the art, and such domains are considered to be single variable domains.
(41) Stabilizing mutation refers to a change of an amino acid residue in a polypeptide sequence that increases the thermal thermostability of said polypeptide. Increased thermostability may be reflected in a melting temperature (Tm) increase of, for example, between 1 and 50 C. CD4 domains with stabilizing mutations include SEQ ID NOs: 5-21.
(42) A variant sequence substantially retains the biological characteristics of the unmodified protein. In the case of an antibody sequence disclosed herein, the VH or VL (or HC or LC) sequence may be a variant sequence with up to 10 amino acid substitutions, additions or deletions. For example, the variant sequence may have up to 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitution(s), addition(s) or deletion(s). The sequence variation may exclude one or more or all of the CDRs, for example the CDRs are the same as the VH or VL (or HC or LC) sequence and the variation is in the remaining portion of the VH or VL (or HC or LC) sequence, so that the CDR sequences are fixed and intact.
(43) V3 loop region, V3/glycan or V3 as used herein refers to the third variable region (V3) of HIV gp120. Comparison of predicted amino acid sequences from several different isolates has shown that sequence heterogeneity of gp120 is clustered in five variable regions (designated V1, V2, V3, V4, and V5.) The V3 region contains post-translational modifications, such as glycosylation, and is essential for viral infectivity. The V3 region, although only 35 amino acids long, exhibits considerable sequence variability. Additionally, variability in potential N-linked glycosylation sites allow for further variability in the variable regions of gp120. Together, the V3 region and the N-linked glycosylation sites within and adjacent to the region are understood to comprise the V3 loop region, V3/glycan or V3 as used herein. For example, one site of glycosylation (e.g., oligomannose such as Man-5 to Man-9) is centered on amino acid residue N332 of gp120. Other sites of potential N-linked glycosylation within and adjacent to the V3 loop region include K295, N301, N386, N392 of gp120. The V3 loop is generally considered to be in the region between cysteine residues C296 and C331 of gp120, while some N-linked glycosylation sites are located directly adjacent to the V3 loop. The V3 loop comprises a highly conserved tetrapeptide sequence, GPGR (residues 312 to 315) (Ivanhoff et al., Virology, 187 (2) 1992). HIV-1 cellular entry depends on the interaction of the V3 loop region with an HIV co-receptor, commonly CCR5 or CXCR4. The V3 loop comprises: (i) the base (residues 296-299), (ii) the stem (residues 300-303 and 321-326), and (iii) the crown (residues 304-320) (Friedrich et al., Nature Communications 12, 6705 (2021)). A consensus sequence of the V3 region of gp120 (Milich et al., J Virol., 67 (9): 5623-5634 (1993)) is provided below:
(44) TABLE-US-00003 (SEQIDNO:361) CTRPNNNTRKSIHIGPGRAFYTTGEIIGDIRQAHC
It is understood that the consensus sequence describes the highest frequency of residues emerging on each position of this region across multiple subtypes, but that the V3 loop region of a particular strain may exhibit sequence variability.
(45) A V3-bNAb or anti-V3 bNAb is a bNAb that binds within the V3 loop region. A V3-bNAb may also be referred to herein as an anti-V3 antibody. A V3-bNAb may bind the N332 glycan in the V3 loop region and/or other N-linked glycosylation sites within and adjacent to the V3 loop region.
STATEMENT OF THE INVENTION
(46) An antigen binding protein of the invention binds to the Human Immunodeficiency Virus (HIV) envelope protein. In particular, the antigen binding protein binds to HIV envelope surface glycoprotein 120 (gp120) and is, therefore, also referred to herein as an anti-gp120 binding protein. The anti-gp120 binding protein of the invention binds to at least two different epitopes on gp120, including the V3 loop region (V3) and the CD4 binding site (CD4bs) of gp120.
(47) Bispecific molecules of the invention that bind to the V3 loop region and the CD4bs of gp120 have been shown to effectively neutralize HIV and exhibit significantly better anti-viral activity than monospecific molecules that only bind to the V3 loop region or the CD4bs of gp120, and mixtures of these monospecific molecules. Without being bound by any particular theory, we postulate that the bispecific molecules of the invention bind the two different epitopes in the same or neighboring HIV envelope protein trimers at the same time, such that the bispecific molecules achieve stronger binding (increased avidity) to the HIV envelope proteins. This may be as a result of the high local concentration of the bispecific molecules' binding sites (paratopes) being pre-positioned around their target binding sites (epitopes) on the HIV envelope compared to their monospecific counterparts, which in turn leads to stronger anti-viral activity.
(48) Binding to the CD4 Binding Site (CD4bs) of HIV Gp120
(49) The antigen binding protein of the invention comprises one or more paratopes that bind to the CD4bs of HIV gp120. Binding domains comprising such paratopes may be include by CD4 domains, as well as other anti-CD4bs domains, including those of anti-CD4bs antibodies and CD4bs-binding fragments thereof. Non-Ig constructs that bind to CD4bs are also part of the invention, such as single chain variable fragments (scFvs). In particular, non-Ig constructs such as scFv comprising one or more CDRs, preferably the three light chain CDRs or the three heavy chain CDRs, or a set of six CDRs of such anti-CD4bs antibodies are also part of the invention.
(50) In an embodiment of the invention, the antigen binding protein of the invention comprises an anti-CD4bs antibody or CD4bs-binding fragment thereof, wherein such antibody or fragment thereof comprises a paratope that binds to the CD4bs of HIV gp120. In a further embodiment, the anti-CD4bs antibody is selected from the group consisting of: b12, HJ16, CH103-106, VRCO1-03, VRC-PG04, VRC-PG04b, VRC-CH30-34, 3BNC117, 3BNC60, NIH45-46, 12A12, 12A21, 8ANC131, 8ANC134, 1NC9, and 1B2530.
(51) In an alternative or additional embodiment of the invention, a paratope that binds to the CD4bs of HIV gp120 is formed by a polypeptide domain that binds to the CD4bs of HIV gp120. In a more particular embodiment, the polypeptide domain is a CD4 domain.
(52) CD4 Domains
(53) CD4 domains of the invention include SEQ ID NOs: 1-21.
(54) In an embodiment of the invention, the CD4 domain is a CD4 D1 domain. In an embodiment, the CD4 domain is a human CD4 domain. CD4 D1 domains include human wild-type D1 (SEQ ID NO:3), mD1.22 (SEQ ID NO:4) also known as D1m, and further variants of mD1.22 (SEQ ID NOs: 5-21).
(55) In an embodiment of the invention, the CD4 domain is a CD4 D1D2 domain. In an embodiment, the CD4 domain is a human CD4 D1D2 domain. CD4 D1D2 domains include human wild-type D1D2 (SEQ ID NO:1) and mD1.22-D2 (SEQ ID NO:2).
(56) In an aspect of the invention, a stabilized CD4 domain is provided. In an embodiment of the invention, a stabilized CD4 D1 domain is provided. In an embodiment, the CD4 domain is thermally stable, i.e., thermostable. In an embodiment, the CD4 domain is a thermostable CD4 D1 domain.
(57) In an embodiment of the invention, the CD4 domain comprises one or more stabilizing mutations. In an embodiment, the stabilizing mutations are in the CD4 D1 domain. In an embodiment, the CD4 D1 domain comprises one or more mutations selected from the group consisting of: K8C, K8I, K8V, T11C, E13C, K21C, Q25E, H27C, H27D, G38C, N52W, R58N, R58T, R58V, L61M, G65C, 170C, K72C, E87G, E91H, E91Q, and G99C. In an embodiment, the CD4 D1 domain comprises K8I. In an embodiment, the CD4 D1 domain comprises K8V. In an embodiment, the CD4 D1 domain comprises T1C and K72C. In an embodiment, the CD4 D1 domain comprises K8C and G99C.
(58) CD4 domains of the invention comprising novel and inventive stabilizing mutations include SEQ ID NOs: 5-21.
(59) Increased thermostability may be reflected in a melting temperature (Tm) increase of, for example, between 1 and 50 C.; in particular between 1 and 30 C.; in particular between 1 and 25 C., in particular between 1 and 21 C., more particularly between 5 and 21 C. The Tm increase is determined by measuring the Tm of the CD4 domain(s) comprising one or more stabilizing mutations and subtracting the Tm of the corresponding CD4 domain(s) without said mutation(s). For example, measuring the Tm of a stabilized CD4 D1 domain and subtracting the Tm of the wild-type CD4 D1 domain. In an embodiment, the Tm increase is about 8 C. In an embodiment, the Tm increase is about 9 C. In an embodiment, the Tm increase is about 12 C. In an embodiment, the Tm increase is about 21 C.
(60) In an embodiment, the Tm of the CD4 domain is above 70 C. In an embodiment, the Tm of the CD4 domain is between 70 C. and 95 C. In an embodiment, the Tm of the CD4 domain is between 75 C. and 95 C. In an embodiment, the Tm of the CD4 domain is between 75 C. and 91 C. In an embodiment, the Tm of the CD4 domain is about 76 C., about 77 C., about 78 C., about 79 C., about 80 C., about 81 C., about 82 C., about 83 C., about 84 C., about 85 C., about 86 C., about 87 C., about 88 C., about 89 C., or about 90 C. In an embodiment, the Tm of the CD4 domain is about 90 C. In an embodiment, the Tm of the CD4 domain is about 89 C.
(61) Tm may be determined by routine methods known in the art or as set out in the Examples. In an embodiment, Tm is determined using the Prometheus System (NanoTemper, Mnchen Germany).
(62) Binding to the V3 Loop Region of HIV Gp120
(63) The antigen binding protein of the invention comprises one or more paratopes that bind to the V3 loop region of HIV gp120. Binding domains comprising such paratopes include an anti-V3 bNAb or a V3-binding fragment thereof, as well as a non-Ig construct that binds to V3.
(64) An antigen binding protein of the invention may comprise heavy chain CDRs (CDRH1, CDRH2, and CDRH3) as set out in any row of Table 1. An antigen binding protein of the invention may comprise light chain CDRs (CDRL1, CDRL2, and CDRL3) as set out in any row of Table 1. An antigen binding protein of the invention may comprise a set of six CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) as set out in any row of Table 1.
(65) An antigen binding protein of the invention may comprise heavy chain CDRs (CDRH1, CDRH2, and CDRH3) of any one of PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12. An antigen binding protein of the invention may comprise light chain CDRs (CDRL1, CDRL2, and CDRL3) of any one of PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12. An antigen binding protein of the invention may comprise a set of six CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) of any one of PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12.
(66) An antigen binding protein of the invention may comprise a VH domain as set out in Table 2. An antigen binding protein of the invention may comprise a VL domain as set out in Table 2. An antigen binding protein of the invention may comprise a pair of variable domains (a VH and a VL) as set out in any row of Table 2.
(67) Anti-V3 bNAbs
(68) An antigen binding protein of the invention may comprise an anti-V3 bNAb or a V3-binding fragment thereof. An anti-V3 antibody includes an antibody comprising a set of CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) as set out in any row of Table 1.
(69) TABLE-US-00004 TABLE 1 SEQ ID NOs for the complementarity determining regions (CDRs) of broadly neutralizing antibodies (bNAbs) 1-6 CDRH1 CDRH2 CDRH3 CDRL1 CDRL2 CDRL3 bNAb1 22 23 24 25 26 27 bNAb2 28 29 30 31 32 33 bNAb3 34 35 36 37 38 39 bNAb4 40 41 42 43 44 45 bNAb5 46 47 48 49 50 51 bNAb6 52 53 54 55 56 57 bNAb7 159 160 161 162 163 164 bNAb8 165 166 167 168 169 170 bNAb9 171 172 173 174 175 176 bNAb10 177 178 179 180 181 182 bNAb11 183 184 185 186 187 188 bNAb12 189 190 191 192 193 194 bNAb13 195 196 197 198 199 200 bNAb14 201 202 203 204 205 206 bNAb15 207 208 209 210 211 212 bNAb16 213 214 215 216 217 218 bNAb17 219 220 221 222 223 224 bNAb18 225 226 227 228 229 230 bNAb19 231 232 233 234 235 236 bNAb20 237 238 239 240 241 242 bNAb21 243 244 245 246 247 248 bNAb23 249 250 251 252 253 254
(70) In a particular embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises the CDRs of bNAb1. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a CDRH1 of SEQ ID NO:22, a CDRH2 of SEQ ID NO:23, a CDRH3 of SEQ ID NO: 24, a CDRL1 of SEQ ID NO: 25, a CDRL2 of SEQ ID NO:26 and a CDRL3 of SEQ ID NO:27.
(71) An anti-V3 bNAb may be an antibody comprising a pair of variable domains (a VH and a VL) as set out in any row of Table 2.
(72) An anti-V3 bNAb may be an antibody comprising a heavy chain (HC), with or without M428L/N434S (EU numbering) LS mutations, and a light chain (LC) as set out in any row of Table 2. In an embodiment, the HC comprises LS.
(73) TABLE-US-00005 TABLE 2 SEQ ID NOs for the variable regions (VH and VL) and the heavy chains (HC) and light chains (LC) of bNAbs 1-6. LS refers to M428L/N434S (EU numbering) mutations in the Fc portion of the bNAb. VH VL HC without LS HC with LS LC bNAb1 58 59 61 62 63 bNAb1* 60* 64* bNAb2 65 66 67 68 69 bNAb3 70 71 72 73 74 bNAb4 75 76 77 78 79 bNAb5 80 81 82 83 84 bNAb6 85 86 87 88 89 bNAb7 255 256 257 258 259 bNAb8 260 261 262 263 264 bNAb9 265 266 267 268 269 bNAb10 270 271 272 273 274 bNAb11 275 276 277 278 279 bNAb12 280 281 282 283 284 bNAb13 285 286 287 288 289 bNAb14 290 291 292 293 294 bNAb15 295 296 297 298 299 bNAb16 300 301 302 303 304 bNAb17 305 306 307 308 309 bNAb18 310 311 312 313 314 bNAb19 315 316 317 318 319 bNAb20 320 321 322 323 324 bNAb21 325 326 327 328 329 bNAb22.sup.# 330 331 332 bNAb23 334 335 336 337 338 *a light chain variant with F32Y in the VL and LC.sup.#a heavy chain variant with {~TC to inserts definition}
(74) In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59 or 60. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO: 65 and a VL domain of SEQ ID NO:66. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO: 70 and a VL domain of SEQ ID NO:71. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:75 and a VL domain of SEQ ID NO: 76. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:80 and a VL domain of SEQ ID NO:81. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:85 and a VL domain of SEQ ID NO:86. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:225 and a VL domain of SEQ ID NO:226. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:260 and a VL domain of SEQ ID NO:261. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:265 and a VL domain of SEQ ID NO: 266. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:270 and a VL domain of SEQ ID NO:271. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:275 and a VL domain of SEQ ID NO:276. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:280 and a VL domain of SEQ ID NO: 281. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:285 and a VL domain of SEQ ID NO:286. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO: 290 and a VL domain of SEQ ID NO:291. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:295 and a VL domain of SEQ ID NO:296. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:300 and a VL domain of SEQ ID NO:301. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:305 and a VL domain of SEQ ID NO: 306. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:310 and a VL domain of SEQ ID NO:311. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:320 and a VL domain of SEQ ID NO:321. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:225 or SEQ ID NO:330 and a VL domain of SEQ ID NO:326. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises a VH domain of SEQ ID NO:334 and a VL domain of SEQ ID NO:335.
(75) In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:61 or 62 and a LC of SEQ ID NO:63 or 64. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO: 62 and a LC of SEQ ID NO:63. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:67 or 68 and a LC of SEQ ID NO:69. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:72 or 73 and a LC of SEQ ID NO:74. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:77 or 78 and a LC of SEQ ID NO:79. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:82 or 83 and a LC of SEQ ID NO: 84. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:87 or 88 and a LC of SEQ ID NO:89 In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:257 or 258 and a LC of SEQ ID NO:259. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:262 or 263 and a LC of SEQ ID NO:264. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO: 267 or 268 and a LC of SEQ ID NO:269. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:272 or 273 and a LC of SEQ ID NO:274. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:277 or 278 and a LC of SEQ ID NO: 279. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:282 or 283 and a LC of SEQ ID NO: 284. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO: 292 or 293 and a LC of SEQ ID NO:294. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO: 297 or 298 and a LC of SEQ ID NO:299. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:302 or 303 and a LC of SEQ ID NO:304. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:307 or 308 and a LC of SEQ ID NO: 309. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:312 or 313 and a LC of SEQ ID NO:314. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO: 317 or 318 and a LC of SEQ ID NO:319. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:322 or 323 and a LC of SEQ ID NO:324. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:327, 328, 331 or 332 and a LC of SEQ ID NO:329. In an embodiment, the anti-V3 antibody comprises a HC of SEQ ID NO:336 or 337 and a LC of SEQ ID NO: 329.
(76) Anti-V3 bNAbs known in the art include PGT121-123, PGT125-131, PGT135-137, DH270.6, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12, 438-B11, 447-52D, BG18, DH270.6, ePGT121v1, ePGT121v2, ePGT121v3, EPTC112, and F425-B4e8. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises the six CDRs of any one of PGT121-123, PGT125-131, PGT135-137, DH270.6, QA013.2, 10-1074, 2G12, 438-B11, 447-52D, BG18, DH270.6, ePGT121v1, ePGT121v2, ePGT121v3, EPTC112, and F425-B4e8. In an embodiment, the anti-V3 antibody or V3-binding fragment thereof, comprises the VH domain and the VL domain any one of PGT121-123, PGT125-131, PGT135-137, DH270.6, QA013.2, 10-1074, 2G12, 438-B11, 447-52D, BG18, DH270.6, ePGT121v1, ePGT121v2, ePGT121v3, EPTC112, and F425-B4e8.
(77) An antigen binding protein of the invention may comprise an anti-V3 scFv of any one of the aforementioned anti-V3 bNAbs. In an embodiment, the scFv comprises a VH and VL pair as set out in Table 2. In an embodiment, the scFv comprises a VH and VL pair of any one of PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12. In an embodiment, the C-terminus of the VH domain is attached directly or via a linker to the N-terminus of the VL domain. In an embodiment, the C-terminus of the VL domain is attached directly or via a linker to the N-terminus of the VH domain. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:65 and a VL domain of SEQ ID NO:66. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:70 and a VL domain of SEQ ID NO: 71. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:75 and a VL domain of SEQ ID NO: 76. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:80 and a VL domain of SEQ ID NO:81. In an embodiment, the scFv comprises a VH domain of SEQ ID NO: 85 and a VL domain of SEQ ID NO:86.
(78) An anti-V3 scFv may be fused to an Fc domain. In an embodiment, the scFv is fused to a human Fc domain directly or via a linker (scFv-Fc).
(79) Linkers
(80) Examples of suitable linkers include amino acid sequences that are from 1 amino acid to 150 amino acids in length. In particular, from 1 to 140 amino acids, from 1 to 130 amino acids, from 1 to 120 amino acids, from 1 to 110 amino acids, from 1 to 100 amino acids, from 1 to 90 amino acids, from 1 to 80 amino acids, from 1 to 70 amino acids, from 1 to 60 amino acids, from 1 to 50 amino acids, from 1 to 40 amino acids, from 1 to 30 amino acids, from 1 to 20 amino acids, from 1 to 10 amino acids, from 5 to 30 amino acids.
(81) In an embodiment, the linker is an amino acid sequence from 5 to 30 amino acids in length. In an embodiment, the linker is an amino acid sequence as set forth in any one of SEQ ID NOs: 90 to 95. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO: 90. In an embodiment, the linker is a multimer of the amino acid sequence as set forth in SEQ ID NO:90. In an embodiment, the linker is [SEQ ID NO:90].sup.n, wherein n is an integer from 1 to 6. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:91. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:92. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:93. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:94. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:95.
(82) Any of the aforementioned linkers may be incorporated into an antigen binding protein of the invention. In particular, any of the aforementioned linkers may be used to join a domain within the antigen binding protein to another domain within the antigen binding protein. In particular, any of the aforementioned linkers may be used to join a domain within the antigen binding protein that binds to the CD4-binding site of HIV gp120 to another domain within the antigen binding protein that binds to the V3 loop region of HIV gp120. Further, any of the aforementioned linkers may be used to join a CD4 domain as disclosed herein to a bNAb as disclosed herein. In an embodiment, the linker is an amino acid sequence as set forth in any one of SEQ ID NOs: 90 to 95. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:90.
(83) In an embodiment, a linker is used to join the C-terminus of a CD4 domain to the N-terminus of a bNAb heavy chain variable domain. In an embodiment, a linker is used to join the C-terminus of a CD4 domain to the N-terminus of a bNAb light chain variable domain. In an embodiment, a linker is used to join the C-terminus of a CD4 domain to the N-terminus of a bNAb heavy chain variable domain and a linker is used to join the C-terminus of a CD4 domain to the N-terminus of a bNAb light chain variable domain. In an embodiment, a linker is used to join the C-terminus of a CD4 domain to the N-terminus of a bNAb heavy chain variable domain and an identical linker is used to join the C-terminus of a CD4 domain to the N-terminus of a bNAb light chain variable domain. In an embodiment, the linker is an amino acid sequence as set forth in any one of SEQ ID NOs: 90 to 95. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:90.
(84) In an embodiment, a linker is used to join the N-terminus of a CD4 domain to the C-terminus of a bNAb heavy chain. In an embodiment, a linker is used to join the N-terminus of a CD4 domain to the C-terminus of a bNAb heavy chain variable domain. In an embodiment, a linker is used to join the N-terminus of a CD4 domain to the C-terminus of a bNAb light chain. In an embodiment, a linker is used to join the N-terminus of a CD4 domain to the C-terminus of a bNAb light chain variable domain. In an embodiment, a linker is used to join the N-terminus of a CD4 domain to the C-terminus of an Fc domain. In an embodiment, the linker is an amino acid sequence as set forth in any one of SEQ ID NOs: 90 to 95. In an embodiment, the linker is an amino acid sequence as set forth in SEQ ID NO:90.
(85) In an embodiment of the invention, the domain of the antigen binding protein that binds to the CD4-binding site of HIV gp120 is joined directly to another domain within the antigen binding protein that binds to the V3 loop region of HIV gp120, i.e., a linker is not used. In an embodiment, a CD4 domain as disclosed herein is joined directly to a bNAb as disclosed herein.
(86) Any of the aforementioned linkers may be used to join a VH and VL pair as disclosed herein to form a scFv. In an embodiment, the linker between the VH domain and the VL domain of the scFv is selected from the group consisting of SEQ ID NOs: 90-95. In a particular embodiment, the linker between the VH domain and the VL domain of the scFv is SEQ ID NO:93.
(87) Any of the aforementioned linkers may be used to join a scFv as disclosed herein to an Fc domain. In an embodiment, the scFv is fused to a human Fc via a linker selected from the group consisting of SEQ ID NO:90-95. In an embodiment, the scFv is fused to a human Fc via a linker of SEQ ID NO:91.
(88) Bispecific Molecules
(89) A bispecific molecule of the invention comprises one or more paratopes that bind to the CD4bs of HIV gp120 and one or more paratopes that bind to the V3 loop region of HIV gp120.
(90) Paratopes that bind to the CD4bs of HIV gp120 may be formed by CD4 domains disclosed herein, as well as other CD4bs-binding domains disclosed herein, including those of anti-CD4bs antibodies and CD4bs-binding fragments thereof, and non-Ig constructs that bind to CD4bs.
(91) Paratopes that bind to the V3 loop region of HIV gp120 may be formed by anti-V3 antibodies and V3-binding fragments thereof disclosed herein, as well as non-Ig constructs that bind to V3 disclosed herein.
(92) In an embodiment, the bispecific molecule comprises a paratope that binds to the CD4bs of HIV gp120 that is formed by an anti-CD4bs antibody or CD4bs-binding fragment thereof and a paratope that binds to the V3 loop region of HIV gp120 that is formed by an anti-V3 antibody or a V3-binding fragment thereof.
(93) In an embodiment, the bispecific molecule comprises an anti-CD4bs antibody or CD4bs-binding fragment thereof and an anti-V3 antibody or a V3-binding fragment thereof.
(94) In an embodiment, the bispecific molecule comprises a CD4 domain and a paratope that binds to the V3 loop region of HIV gp120 that is formed by an anti-V3 antibody or a V3-binding fragment thereof.
(95) In an embodiment, the bispecific molecule comprises a CD4 domain and an anti-V3 antibody or a V3-binding fragment thereof.
(96) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody or a V3-binding fragment thereof.
(97) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO: 11 and an anti-V3 antibody or a V3-binding fragment thereof.
(98) In an embodiment, the bispecific molecule comprises a CD4 domain and an anti-V3 antibody, selected from the group consisting of: PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12, or a V3-binding fragment thereof.
(99) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody selected from the group consisting of: PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12, or a V3-binding fragment thereof.
(100) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO: 11 and an anti-V3 antibody selected from the group consisting of: PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12, or a V3-binding fragment thereof.
(101) In an embodiment, the bispecific molecule comprises a CD4 domain and an anti-V3 antibody comprising a set of CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) as set out in any row of Table 1, or a V3-binding fragment thereof.
(102) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a set of CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) as set out in any row of Table 1, or a V3-binding fragment thereof.
(103) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a set of CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) as set out in any row of Table 1 or a V3-binding fragment thereof.
(104) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO: 11 and an anti-V3 antibody comprising a CDRH1 of SEQ ID NO:22, a CDRH2 of SEQ ID NO:23, a CDRH3 of SEQ ID NO:24, a CDRL1 of SEQ ID NO: 25, a CDRL2 of SEQ ID NO:26 and a CDRL3 of SEQ ID NO:27, or a V3-binding fragment thereof.
(105) In an embodiment, the bispecific molecule comprises a CD4 domain and an anti-V3 antibody comprising a pair of variable domains (a VH and a VL) as set out in any row of Table 2, or a V3-binding fragment thereof.
(106) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a pair of variable domains (a VH and a VL) as set out in any row of Table 2, or a V3-binding fragment thereof.
(107) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO: 11 and an anti-V3 antibody comprising a pair of variable domains (a VH and a VL) as set out in any row of Table 2, or a V3-binding fragment thereof.
(108) In an embodiment, the bispecific molecule comprises a CD4 domain and an anti-V3 antibody or a V3-binding fragment thereof comprising a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59 or 60.
(109) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59 or 60.
(110) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO: 59 or 60.
(111) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:65 and a VL domain of SEQ ID NO:66.
(112) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO: 11 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:65 and a VL domain of SEQ ID NO: 66.
(113) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a VH domain of SEQ ID NO: 70 and a VL domain of SEQ ID NO:71.
(114) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:70 and a VL domain of SEQ ID NO: 71.
(115) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:75 and a VL domain of SEQ ID NO:76.
(116) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:75 and a VL domain of SEQ ID NO: 76.
(117) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:80 and a VL domain of SEQ ID NO:81.
(118) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:80 and a VL domain of SEQ ID NO: 81.
(119) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:85 and a VL domain of SEQ ID NO:86.
(120) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a VH domain of SEQ ID NO:85 and a VL domain of SEQ ID NO: 86.
(121) An anti-V3 antibody as described above may be an antibody comprising a heavy chain (HC), with or without M428L/N434S (EU numbering) LS mutations. In an embodiment, the HC comprises LS.
(122) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:61 or 62 and a LC of SEQ ID NO: 63 or 64.
(123) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a HC of SEQ ID NO:61 or 62 and a LC of SEQ ID NO:63 or 64.
(124) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:62 and a LC of SEQ ID NO: 63.
(125) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a HC of SEQ ID NO:62 and a LC of SEQ ID NO:63.
(126) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:67 or 68 and a LC of SEQ ID NO: 69.
(127) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO: 11 and an anti-V3 antibody comprising a HC of SEQ ID NO:67 or 68 and a LC of SEQ ID NO:69.
(128) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:72 or 73 and a LC of SEQ ID NO: 74.
(129) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a HC of SEQ ID NO:72 or 73 and a LC of SEQ ID NO:74.
(130) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:77 or 78 and a LC of SEQ ID NO: 79.
(131) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a HC of SEQ ID NO:77 or 78 and a LC of SEQ ID NO:79.
(132) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:82 or 83 and a LC of SEQ ID NO: 84.
(133) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a HC of SEQ ID NO:82 or 83 and a LC of SEQ ID NO:84.
(134) In an embodiment, the bispecific molecule comprises a CD4 domain of any one of SEQ ID NOs: 1-21 and an anti-V3 antibody comprising a HC of SEQ ID NO:87 or 88 and a LC of SEQ ID NO: 89
(135) In an embodiment, the bispecific molecule comprises a CD4 domain of SEQ ID NO:11 and an anti-V3 antibody comprising a HC of SEQ ID NO:87 or 88 and a LC of SEQ ID NO:89.
(136) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and two copies of a CD4 domain as disclosed herein, wherein the C-terminus of one CD4 domain is attached directly or by a linker to the N-terminus of one of the anti-V3 bNAb heavy chains and the C-terminus of the other copy of the CD4 domain is attached directly or by a linker to the N-terminus of the other anti-V3 bNAb heavy chain.
(137) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and two copies of a CD4 domain as disclosed herein, wherein the C-terminus of one CD4 domain is attached directly or by a linker to the N-terminus of one of the anti-V3 bNAb light chains and the C-terminus of the other copy of the CD4 domain is attached directly or by a linker to the N-terminus of the other anti-V3 bNAb light chain.
(138) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and four copies of a CD4 domain as disclosed herein, wherein the C-terminus of the first CD4 domain is attached directly or by a linker to the N-terminus of one of the anti-V3 bNAb heavy chains, the C-terminus of the second CD4 domain is attached directly or by a linker to the N-terminus of the other anti-V3 bNAb heavy chains, the third CD4 domain is attached directly or by a linker to the N-terminus of one of the anti-V3 bNAb light chains, and the fourth CD4 domain is attached directly or by a linker to the N-terminus of the other anti-V3 bNAb light chains.
(139) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and two copies of a CD4 domain as disclosed herein, wherein the N-terminus of the first CD4 domain is attached directly or by a linker to the C-terminus of one of the anti-V3 bNAb heavy chains and the N-terminus of the other copy of the CD4 domain is attached directly or by a linker to the C-terminus of the other anti-V3 bNAb heavy chain.
(140) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and two copies of a CD4 domain of SEQ ID NO: 11, wherein the C-terminus of one CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb heavy chains and the C-terminus of the other copy of the CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb heavy chain.
(141) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and two copies of a CD4 domain of SEQ ID NO: 11, wherein the C-terminus of one CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb light chains and the C-terminus of the other copy of the CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb light chain.
(142) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and four copies of a CD4 domain of SEQ ID NO:11, wherein the C-terminus of the first CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb heavy chains, the C-terminus of the second CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb heavy chains, the third CD4 domain is by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb light chains, and the fourth CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb light chains.
(143) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb as disclosed herein and two copies of a CD4 domain of SEQ ID NO: 11, wherein the N-terminus of the first CD4 domain is attached by a linker of SEQ ID NO: 90 to the C-terminus of one of the anti-V3 bNAb heavy chains and the N-terminus of the other copy of the CD4 domain is attached by a linker of SEQ ID NO: 90 to the C-terminus of the other anti-V3 bNAb heavy chain.
(144) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb comprising a VH of SEQ ID NO:58 and a VL of SEQ ID NO:59, and two copies of a CD4 domain of SEQ ID NO: 11, wherein the C-terminus of one CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb heavy chains and the C-terminus of the other copy of the CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb heavy chain.
(145) In an embodiment, the bispecific molecule an anti-V3 bNAb comprising a VH of SEQ ID NO: 58 and a VL of SEQ ID NO:59, and two copies of a CD4 domain of SEQ ID NO: 11, wherein the C-terminus of one CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb light chains and the C-terminus of the other copy of the CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb light chain.
(146) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb comprising a VH of SEQ ID NO:58 and a VL of SEQ ID NO:59, and four copies of a CD4 domain of SEQ ID NO: 11, wherein the C-terminus of the first CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb heavy chains, the C-terminus of the second CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb heavy chains, the third CD4 domain is by a linker of SEQ ID NO: 90 to the N-terminus of one of the anti-V3 bNAb light chains, and the fourth CD4 domain is attached by a linker of SEQ ID NO: 90 to the N-terminus of the other anti-V3 bNAb light chains.
(147) In an embodiment, the bispecific molecule comprises an anti-V3 bNAb comprising a VH of SEQ ID NO:58 and a VL of SEQ ID NO:59, and two copies of a CD4 domain of SEQ ID NO: 11, wherein the N-terminus of the first CD4 domain is attached by a linker of SEQ ID NO: 90 to the C-terminus of one of the anti-V3 bNAb heavy chains and the N-terminus of the other copy of the CD4 domain is attached by a linker of SEQ ID NO: 90 to the C-terminus of the other anti-V3 bNAb heavy chain.
(148) In an embodiment, the bispecific molecule comprises a HC of any one of SEQ ID NOs: 96-107, 116, 117 or 119-135; and a LC of SEQ ID NO:63.
(149) In an embodiment, the bispecific molecule comprises a HC of SEQ ID NO:62; and a LC of any one of SEQ ID NOs: 108-115 and 118.
(150) In an embodiment, the bispecific molecule comprises a HC of SEQ ID NO:68; and a LC of SEQ ID NO: 142 or 143.
(151) In an embodiment, the bispecific molecule comprises a HC of any one of SEQ ID NOs: 136-141; and a LC of SEQ ID NO:69.
(152) In an embodiment, the bispecific molecule comprises a HC of SEQ ID NO: 144 or 145; and a LC of SEQ ID NO:74.
(153) In an embodiment, the bispecific molecule comprises a HC of SEQ ID NO: 146 or 147; and a LC of SEQ ID NO:79.
(154) In an embodiment, the bispecific molecule comprises a HC of SEQ ID NO: 148 or 149; and a LC of SEQ ID NO:84.
(155) In an embodiment, the bispecific molecule comprises a HC of SEQ ID NO: 150 or 151; and a LC of SEQ ID NO:89.
(156) In an embodiment, the bispecific molecule consists of two heavy chains and two light chains, wherein the heavy chain is at least 95% identical to SEQ ID NO: 121 and the light chain that is at least 95% identical to SEQ ID NO:63.
(157) In an embodiment, the bispecific molecule consists of two heavy chains and two light chains, wherein the heavy chain is at least 96% identical to SEQ ID NO: 121 and the light chain that is at least 96% identical to SEQ ID NO:63.
(158) In an embodiment, the bispecific molecule consists of two heavy chains and two light chains, wherein the heavy chain is at least 97% identical to SEQ ID NO: 121 and the light chain that is at least 97% identical to SEQ ID NO:63.
(159) In an embodiment, the bispecific molecule consists of two heavy chains and two light chains, wherein the heavy chain is at least 98% identical to SEQ ID NO:121 and the light chain that is at least 98% identical to SEQ ID NO:63.
(160) In an embodiment, the bispecific molecule consists of two heavy chains and two light chains, wherein the heavy chain is at least 99% identical to SEQ ID NO: 121 and the light chain that is at least 99% identical to SEQ ID NO:63.
(161) In an embodiment, the bispecific molecule consists of two heavy chains of SEQ ID NO: 121 and two light chains of SEQ ID NO:63.
(162) An antigen binding protein of the invention may comprise an anti-V3 scFv of any one of the aforementioned anti-V3 bNAbs. In an embodiment, the scFv comprises a VH and VL pair as set out in Table 2. In an embodiment, the scFv comprises a VH and VL pair of any one of PGT121-123, PGT125-131, PGT135-137, QA013.2, 10-1074, 10-1074LS, PGT121.414.LS and 2G12. In an embodiment, the C-terminus of the VH domain is attached directly or via a linker to the N-terminus of the VL domain. In an embodiment, the C-terminus of the VL domain is attached directly or via a linker to the N-terminus of the VH domain. In an embodiment, the linker between the VH domain and the VL domain of the scFv is selected from the group consisting of SEQ ID NOs: 90-95. In an embodiment, the linker between the VH domain and the VL domain of the scFv is SEQ ID NO:93.
(163) In an embodiment, the scFv comprises a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:65 and a VL domain of SEQ ID NO: 66. In an embodiment, the scFv comprises a VH domain of SEQ ID NO: 70 and a VL domain of SEQ ID NO:71. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:75 and a VL domain of SEQ ID NO:76. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:80 and a VL domain of SEQ ID NO:81. In an embodiment, the scFv comprises a VH domain of SEQ ID NO:85 and a VL domain of SEQ ID NO:86. In an embodiment, a linker of SEQ ID NO:93 joins the VH domain and the VL domain of the scFv. In an embodiment, a linker of SEQ ID NO:93 joins the C terminal of the VH domain to the N terminal of the VL domain to form the scFv. In an embodiment, a linker of SEQ ID NO:93 joins the C terminal of the VL domain to the N terminal of the VH domain to form the scFv.
(164) An anti-V3 scFv may be fused to an Fc domain. In an embodiment, the scFv is fused to a human Fc domain directly or via a linker (scFv-Fc). In an embodiment, the C-terminus of the scFv is fused to the N-terminus of a human Fc domain via a linker selected from the group consisting of SEQ ID NOs: 90-95. In an embodiment, the N-terminus of the scFv is fused to the C-terminus of a human Fc domain via a linker selected from the group consisting of SEQ ID NOs: 90-95. In an embodiment, the scFv is fused to a human Fc domain via a linker of SEQ ID NO: 91. In an embodiment, the Fc domain comprises a half-life extending mutation. In an embodiment the half-life extending mutation is LS.
(165) A scFv-Fc may be fused directly or via a linker to a CD4 domain. In an embodiment, the scFv-Fc is fused via a linker selected from the group consisting of SEQ ID NOs: 90-95 to a CD4 domain.
(166) In an embodiment, the antigen binding protein comprises or consists of: (1) a scFv comprising a VH and VL pair as set out in any row of Table 2, wherein the VH and VL domain are joined to form a scFV via a linker selected from the group consisting of SEQ ID NOs: 90-95; (2) a CD4 domain selected from the group consisting of SEQ ID NOs: 1-21; and (3) and Fc domain comprising LS half-life extending mutations; wherein (1), (2) and (3) are joined together in any order directly or via a linker, and wherein each linker is selected from the group consisting of SEQ ID NOs: 90-95.
(167) In an embodiment, the antigen binding protein comprises or consists of: (1) a scFv comprising a VH and VL pair as set out in any row of Table 2, wherein the VH and VL domain are joined to form a scFV via a linker of SEQ ID NO:93; (2) a CD4 domain selected from the group consisting of SEQ ID NOs: 1-21; and (3) and Fc domain comprising LS half-life extending mutations; wherein (1), (2) and (3) are joined together in any order via a linker between each domain, and wherein the linker is SEQ ID NOs: 90.
(168) In an embodiment, the bispecific molecule of the invention comprises a sequence that is at least 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 152-157. In an embodiment, an antigen binding protein of the invention consists of a sequence that is at least 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 152-157. In an embodiment, an antigen binding protein of the invention comprises or consists of SEQ ID NO: 152. In an embodiment, an antigen binding protein of the invention comprises or consists of SEQ ID NO:153. In an embodiment, an antigen binding protein of the invention comprises or consists of SEQ ID NO: 154. In an embodiment, an antigen binding protein of the invention comprises or consists of SEQ ID NO: 155. In an embodiment, an antigen binding protein of the invention comprises or consists of SEQ ID NO: 156. In an embodiment, an antigen binding protein of the invention comprises or consists of SEQ ID NO: 157.
(169) Production Methods
(170) Antigen binding proteins may be prepared by any of a number of conventional techniques. For example, antigen binding proteins may be purified from cells that naturally express them (e.g., an antibody can be purified from a hybridoma that produces it), or produced in recombinant expression systems.
(171) A number of different expression systems and purification regimes can be used to generate the antigen binding proteins of the invention. Generally, host cells are transformed with a recombinant expression vector encoding the desired antigen binding protein. The expression vector may be maintained by the host as a separate genetic element or integrated into the host chromosome depending on the expression system. A wide range of host cells can be employed, including Prokaryotes (including Gram-negative or Gram-positive bacteria, for example Escherichia coli, Bacilli sp., Pseudomonas sp., Corynebacterium sp.), Eukaryotes including yeast (for example Saccharomyces cerevisiae, Pichia pastoris), fungi (for example Aspergillus sp.), or higher Eukaryotes including insect cells and cell lines of mammalian origin (for example, CHO, NS0, PER.C6, HEK293, HeLa).
(172) The host cell may be an isolated host cell. The host cell is usually not part of a multicellular organism (e.g., plant or animal). The host cell may be a non-human host cell.
(173) Appropriate cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian host cells are known in the art.
(174) The cells can be cultured under conditions that promote expression of the antigen binding protein using a variety of equipment such as shake flasks, spinner flasks, and bioreactors. The polypeptide(s) is (are) recovered by conventional protein purification procedures. Protein purification procedures typically consist of a series of unit operations comprised of various filtration and chromatographic processes developed to selectively concentrate and isolate the antigen binding protein. The purified antigen binding protein may be formulated in a pharmaceutically acceptable composition.
(175) Fc Modifications
(176) Fc engineering methods can be applied to modify the functional or pharmacokinetics properties of an antigen binding protein, in particular an antibody. Effector function may be altered by making mutations in the Fc region that increase or decrease binding to Clq or Fc receptors and modify CDC or ADCC activity respectively. Modifications to the glycosylation pattern of an antibody can also be made to change the effector function.
(177) The interaction between the Fc region of an antigen binding protein or antibody and various Fc receptors (FcR), including FcRI (CD64), FcRII (CD32), FcRIII (CD16), FcRn, C1q, and type II Fc receptors is believed to mediate the effector functions of the antigen binding protein or antibody. Significant biological effects can be a consequence of effector functionality. Usually, the ability to mediate effector function requires binding of the antigen binding protein or antibody to an antigen and not all antigen binding proteins or antibodies will mediate every effector function.
(178) Effector function can be assessed in a number of ways including, for example, evaluating ADCC effector function of antibody coated to target cells mediated by Natural Killer (NK) cells via FcRIII, or monocytes/macrophages via FcRI, or evaluating CDC effector function of antibody coated to target cells mediated by complement cascade via Clq. For example, an antigen binding protein of the present invention can be assessed for ADCC effector function in a Natural Killer cell assay. Examples of such assays can be found in Shields et al, 2001, The Journal of Biological Chemistry, Vol. 276, p. 6591-6604; Chappel et al, 1993, The Journal of Biological Chemistry, Vol 268, p. 25124-25131; Lazar et al, 2006, PNAS, 103; 4005-4010.
(179) Examples of assays to determine CDC function include those described in J Imm Meth, 1995, 184:29-38.
(180) The effects of mutations on effector functions (e.g., FcRn binding, FcRs and C1q binding, CDC, ADCML, ADCC, ADCP) can be assessed, e.g., as described in Grevys et al., J Immunol. 2015 Jun. 1; 194 (11): 5497-5508, or Tam et al., Antibodies 2017, 6 (3); Monnet et al., 2014 mAbs, 6:2, 422-436.
(181) Throughout this specification, amino acid residues in Fc regions, in antibody sequences or full-length antigen binding protein sequences, are numbered according to the EU index numbering convention.
(182) The long half-life of IgG antibodies is reported to be dependent on their binding to FcRn. Therefore, substitutions that increase the binding affinity of IgG to FcRn at pH 6.0 while maintaining the pH dependence of the interaction with target, by engineering the constant region, have been extensively studied (Ghetie et al., Nature Biotech. 15:637-640, 1997; Hinton et al., JBC 279:6213-6216, 2004; Dall'Acqua et al., 10 J Immunol 117:1129-1138, 2006). The in-vivo half-life of antigen binding proteins of the present invention may be altered by modification of a heavy chain constant domain or an FcRn binding domain therein.
(183) In adult mammals, FcRn, plays a key role in maintaining serum antibody levels by acting as a protective receptor that binds and salvages antibodies of the IgG isotype from degradation. IgG molecules are endocytosed by endothelial cells and, if they bind to FcRn, are recycled out of the cells back into circulation. In contrast, IgG molecules that enter the cells and do not bind to FcRn and are targeted to the lysosomal pathway where they are degraded.
(184) FcRn is believed to be involved in both antibody clearance and the transcytosis across tissues (see Junghans R. P (1997) Immunol.Res 16. 29-57 and Ghetie et al (2000) Annu. Rev. Immunol. 18, 739-766). Human IgG1 residues determined to interact directly with human FcRn include Ile253, Ser254, Lys288, Thr307, Gln311, Asn434 and His435. Mutations at any of these positions may enable increased serum half-life and/or altered effector properties of antigen binding proteins of the invention.
(185) Antigen binding proteins of the present invention may have amino acid modifications that increase the affinity of the constant domain or fragment thereof for FcRn. Increasing the half-life (i.e., serum half-life) of therapeutic and diagnostic IgG antibodies and other bioactive molecules has many benefits including reducing the amount and/or frequency of dosing of these molecules. In one embodiment, an antigen binding protein of the invention comprises all or a portion (an FcRn binding portion) of an IgG constant domain having one or more of the following amino acid modifications.
(186) For example, with reference to IgG1, M252Y/S254T/T256E (commonly referred to as YTE mutations) and M428L/N434S (commonly referred to as LS mutations) increase FcRn binding at pH 6.0 (Wang et al. 2018). In an embodiment, an antigen binding protein of the invention comprises an Fc domain with the LS mutations. In an embodiment, an antigen binding protein of the invention comprises a bNAb in which the LS mutations are present in both of the heavy chain Fc domains.
(187) Half-life and FcRn binding can also be extended by introducing H433K and N434F mutations (commonly referred to as HN or NHance mutations) (with reference to IgG1) (WO2006/130834).
(188) Additionally, various publications describe methods for obtaining physiologically active molecules with modified half-lives, either by introducing an FcRn-binding polypeptide into the molecules (WO97/43316, U.S. Pat. Nos. 5,869,046, 5,747,035, WO96/32478 and WO91/14438) or by fusing the molecules with antibodies whose FcRn-binding affinities are preserved, but affinities for other Fc receptors have been greatly reduced (WO99/43713), or fusing with FcRn binding domains of antibodies (WO00/09560, U.S. Pat. No. 4,703,039).
(189) Post-Translational Modifications
(190) The skilled person will appreciate that, upon production of an antigen binding protein, such as a bispecific molecule of the invention in a host cell, post-translational modifications may occur. For example, this may include the cleavage of certain leader sequences, the addition of various sugar moieties in various glycosylation patterns, non-enzymatic glycation, deamidation, oxidation, disulfide bond scrambling and other cysteine variants such as free sulfhydryls, racemized disulfides, thioethers and trisulfide bonds, isomerisation, C-terminal lysine clipping, and N-terminal glutamine cyclisation. The present invention encompasses the use of antigen binding proteins that have been subjected to, or have undergone, one or more post-translational modifications. Thus an antigen binding protein of the invention includes an antigen binding protein as defined earlier that has undergone a post-translational modification such as described herein.
(191) Glycation is a post-translational non-enzymatic chemical reaction between a reducing sugar, such as glucose, and a free amine group in the protein, and is typically observed at the epsilon amine of lysine side chains or at the N-Terminus of the protein. Glycation can occur during production and storage only in the presence of reducing sugars.
(192) Deamidation can occur during production and storage, is an enzymatic reaction primarily converting asparagine (N) to iso-aspartic acid (iso-aspartate) and aspartic acid (aspartate) (D) at approximately 3:1 ratio. This deamidation reaction is therefore related to isomerization of aspartate (D) to iso-aspartate. The deamidation of asparagine and the isomerisation of aspartate, both involve the intermediate succinimide. To a much lesser degree, deamidation can occur with glutamine residues in a similar manner. Deamidation can occur in a CDR, in a Fab (non-CDR region), or in the Fc region.
(193) Oxidation can occur during production and storage (i.e., in the presence of oxidizing conditions) and results in a covalent modification of a protein, induced either directly by reactive oxygen species or indirectly by reaction with secondary by-products of oxidative stress. Oxidation happens primarily with methionine residues, but may occur at tryptophan and free cysteine residues. Oxidation can occur in a CDR, in a Fab (non-CDR) region, or in the Fc region.
(194) Disulfide bond scrambling can occur during production and basic storage conditions. Under certain circumstances, disulfide bonds can break or form incorrectly, resulting in unpaired cysteine residues (SH). These free (unpaired) sulfhydryls (SH) can promote shuffling.
(195) The formation of a thioether and racemization of a disulphide bond can occur under basic conditions, in production or storage, through a beta elimination of disulphide bridges back to cysteine residues via a dehydroalanine and persulfide intermediate. Subsequent crosslinking of dehydroalanine and cysteine results in the formation of a thioether bond or the free cysteine residues can reform a disulphide bond with a mixture of D- and L-cysteine.
(196) Trisulfides result from insertion of a sulfur atom into a disulphide bond (Cys-SSS-Cys) and are formed due to the presence of hydrogen sulphide in production cell culture.
(197) N-terminal glutamine (Q) and glutamate (glutamic acid) (E) in the heavy chain and/or light chain is likely to form pyroglutamate (pGlu) via cyclization. Most pGlu formation happens in the production bioreactor, but it can be formed non-enzymatically, depending on pH and temperature of processing and storage conditions. Cyclization of N-terminal Q or E is commonly observed in natural human antibodies.
(198) C-terminal lysine clipping is an enzymatic reaction catalyzed by carboxypeptidases, and is commonly observed in recombinant and natural human antibodies. Variants of this process include removal of lysine from one or both heavy chains due to cellular enzymes from the recombinant host cell. Upon administration to the human subject/patient is likely to result in the removal of any remaining C-terminal lysines.
(199) Pharmaceutical Compositions
(200) Antigen binding proteins as described herein may be incorporated into pharmaceutical compositions for use in the treatment or prevention of HIV infection. In one embodiment, the pharmaceutical composition comprises an antigen binding protein in combination with one or more pharmaceutically acceptable carriers and/or excipients.
(201) Such compositions comprise a pharmaceutically acceptable carrier as known and called for by acceptable pharmaceutical practice.
(202) Pharmaceutical compositions may be administered by injection or continuous infusion (examples include, but are not limited to, intravenous, intraperitoneal, intradermal, subcutaneous, intramuscular, intraocular, and intraportal). In one embodiment, the composition is suitable for intravenous administration. In one embodiment, the composition is suitable for subcutaneous administration.
(203) Pharmaceutical compositions may be suitable for topical administration (which includes, but is not limited to, epicutaneous, inhaled, intranasal or ocular administration) or enteral administration (which includes, but is not limited to, oral, vaginal, or rectal administration).
(204) The pharmaceutical composition may be included in a kit containing the antigen binding protein together with other medicaments, for example dolutegravir or cabotegravir, and/or with instructions for use. For convenience, the kit may comprise the reagents in predetermined amounts with instructions for use. The kit may also include devices used for administration of the pharmaceutical composition.
(205) The terms individual, subject and patient are used herein interchangeably. In one embodiment the subject is a human.
(206) The antigen binding proteins described herein may be used in methods of treatment or prevention of HIV infection and AIDs. The antigen binding proteins described herein may be used in the manufacture of medicaments for the treatment or prevention of HIV infection and AIDs. The antigen binding proteins described may be used in an effective amount for therapeutic, prophylactic or preventative treatment. A therapeutically effective amount of the antigen binding protein described herein is an amount effective to ameliorate or reduce one or more symptoms of HIV infection. A prophylactically effective amount of the antigen binding protein described herein is an amount effective to prevent one or more symptoms of HIV infection.
(207) Combinations
(208) Antigen binding proteins of the present invention may be employed alone or in combination with other therapeutic agents, or a prodrug thereof. Combination therapies according to the present invention thus comprise the administration of an antigen binding protein and the administration of at least one other agent which may be useful in the treatment or prevention of HIV infection and/or AIDS. An antigen binding protein of the present invention and the other therapeutic agent may be formulated and administered together in a single pharmaceutical composition or may be formulated and administered separately. When formulated and administered separately, administration may occur simultaneously or sequentially in any order.
(209) Antigen binding proteins as described herein may be combined with, for example, one or more of an antiretroviral agent, an anti-infective agent, an immunomodulator, and other HIV entry inhibitors.
(210) Antiretroviral agents include Nucleoside Reverse Transcriptase Inhibitors (NRTIs), Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs), Nucleoside Reverse Transcriptase Translocation Inhibitors (NRTTIs), Protease Inhibitors (PIs), Entry Inhibitors (EI), Integrase Strand Transfer Inhibitors (INSTI), Maturation Inhibitors (MIS), and Capsid Inhibitors (CIs).
(211) NRTIs may include, but are not limited to: abacavir, adefovir, adefovir dipivoxil, alovudine, amdoxovir, apricitabine, calanolide A, censavudine, didanosine, elvucitabine, emtricitabine, fozivudine, lamivudine, racivir, stampidine, stavudine, tenofovir disoproxil fumerate, tenofovir alafenamide, todoxil, zalcitabine, and zidovudine.
(212) NNRTIs may include, but are not limited to, HBY 097 (Hoechst/Bayer), capravirine, delaviridine, doravirine, efavirenz, etravirine, immunocal, lersivirine, loviride, nevirapine, oltipraz, and rilpivirine.
(213) NRTTIs include, but are not limited to, islatravir.
(214) PIs may include, but are not limited to, amprenavir, atazanavir, brecanavir, cobicistat, darunavir, fosamprenavir, indinavir, lasinavir, lopinavir, palinavir, nelfinavir, ritonavir, saquinavir, and tipranavir.
(215) EIs are discussed in DRUGS OF THE FUTURE 1999, 24 (12), 1355-1362; CELL, Vol. 9, 243-246, Oct. 29, 1999; and DRUG DISCOVERY TODAY, Vol. 5, No. 5, May 2000, pp. 183-194; and Meanwell et al., Current Opinion in Drug Discovery & Development (2003), 6 (4), 451-461. In particular, the antigen binding proteins of the invention can be utilized in combination with attachment inhibitors, fusion inhibitors, and chemokine receptor antagonists aimed at either the CCR5 or CXCR4 coreceptor. HIV attachment inhibitors are also set forth in U.S. Pat. Nos. 7,354,924 and 7,745,625. EIs may include, but are not limited to, cenicriviroc, enfuvirtide, fostemsavir, ibalizumab, leronlimab, maraviroc, vicriviroc and VIR-576.
(216) INSTIs may include, but are not limited to, bictegravir, cabotegravir, dolutegravir, elvitegravir, and raltegravir. In an embodiment, the INSTI is dolutegravir or cabotegravir. In an embodiment, the INSTI is cabotegravir.
(217) Maturation inhibitors may include, but are not limited to, bevirimat, BMS-955176, GSK3640254, GSK3739937, PA-344 and PA-457. It will be understood that GSK3640254 is a compound as described in Dicker I, Jeffrey J L, Protack T, et al., Antimicrob Agents Chemother. 2022; 66 (1). GSK3739937, also known as VH3739937, is the compound of clinical trial NCT04493684.
(218) Capsid inhibitors may include, but are not limited to, GSK4004280, GSK4011499, and lencapavir.
(219) Anti-infective agents include, but are not limited to, clindamycin with primaquine, daunorubicin, fluconazole, intraconazole, nystatin pastille, ornidyl eflornithine, megestrol acetate, pentamidine isethionate, piritrexim, trimethoprim, trimetrexate, recombinant human erythropoietin, recombinant human growth hormone, spiramycin, testosterone and total enteral nutrition,
(220) Immunomodulators include, but are not limited to, acemannan, alpha-2-interferon, AS-101, bropirimine, CL246,738, FP-21399, gamma interferon, granulocyte macrophage colony stimulating factor, HIV core particle immunostimulant, interleukin-2, immune globulin, IMREG-1, IMREG-2, imuthiol diethyl dithio carbamate, methionine enkephalin, MTP-PE muramyl tripeptide, remune, recombinant soluble human CD4, rCD4-IgG hybrids, SK&F106528, thymopentin, and tumour necrosis factor (TNF).
(221) The antigen binding proteins of the present invention may also be used in combination with agents that induce HIV expression, such as latency reversing agents. Several latency reversing agents include, but are not limited to, the following: histone deacetylase inhibitors (e.g., vorinostat, panobinostat, romidepin), histone crotonyl transferase inhibitors (sodium corotonate), protein kinase C agonists (e.g., bryostatin, ingenol B), disulfiram, TLR7 agonists (e.g., GS-9620), and bromodomain inhibitors (e.g., JQ1, iBET151).
(222) The antigen binding proteins of the present invention may also be used in combination with other agents that induce HIV expression, such as agents for clearance therapy. Several examples of agents for clearance therapy, or of immunological combinations for clearance, include, but are not limited to, the following: bNAbs, CD4-Ig, eCD4-Ig, and dual-affinity re-targeting (DART) proteins.
(223) Antigen binding proteins of the invention may be used in combination with broadly neutralizing HIV-1 antibodies, including 1NC9, 1B2530, 2F5, 2G12, 3NBC60, 3BNC117, 4E10, 8ANC131, 8ANC134, 10-1074, 10-1074LS, 10E8, 12A12, 12A21, b12, CAP206-CH12, CH01-04, CH103-106, elipovimab (formerly known as GS-9722), HJ16, M66.6, N6LS (also known as VRC-HIVMAB091-00-AB and the compound of clinical trial NCT03538626), NIH45-46, PG9, PG16, PGT121-123, PGT125-131, PGT135-137, PGT141-145, PGT121.414.LS, PGT151 2G12, QA013.2, VRC01-03, VRC-PG04, VRC-PG04b, VRC-CH30-34.
(224) Other agents that may be combined with antigen binding proteins of the invention include BIT225, GSK4000422/VH4000422, and S-648414 (the compound of clinical trial NCT04147715).
(225) The scope of combinations of compounds of this invention with HIV agents is not limited to those mentioned above but includes in principle any combination with any pharmaceutical composition useful for the treatment and/or prevention of HIV infection and/or AIDS.
(226) The invention is illustrated by the following clauses: 1. An anti-HIV gp120-binding protein that binds to at least two different epitopes on human immunodeficiency virus (HIV) surface glycoprotein 120 (gp120). 2. The anti-HIV gp120-binding protein of clause 1, wherein one of the at least two epitopes comprises one or more amino acid residues of the CD4-binding site of HIV gp120. 3. The anti-HIV gp120-binding protein of clause 1 or clause 2, wherein one of the at least two epitopes comprises one or more amino acid residues of the V3 loop region (V3) of HIV gp120. 4. The anti-HIV gp120-binding protein of clause 3, wherein one of the at least two epitopes comprises a cluster of mannose glycans centered on N332 of HIV gp120. 5. The anti-HIV gp120-binding protein of any one of the preceding clauses that binds to the CD4-binding site of HIV gp120 and the V3 loop region of HIV gp120. 6. The anti-HIV gp120-binding protein of any one of the preceding clauses, comprising a CD4-domain. 7. The anti-HIV gp120-binding protein of clause 6, wherein the CD4 domain is a CD4 D1 domain or CD4 D1D2 domain. 8. The anti-HIV gp120-binding protein of clause 6 or clause 7, wherein the CD4 domain comprises one or more stabilizing mutations. 9. The anti-HIV gp120-binding protein of clause 8, wherein the CD4 domain has a Tm of between 70 C. and 95 C. 10. The anti-HIV gp120-binding protein of any one of clauses 6 to 9, wherein the CD4 D1 domain comprises one or more mutations selected from the group consisting of: K8C, K8I, K8V, T11C, E13C, K21C, Q25E, H27C, H27D, G38C, N52W, R58N, R58T, R58V, L61M, G65C, 170C, K72C, E87G, E91H, E91Q, and G99C. 11. The anti-HIV gp120-binding protein of clause 9, wherein the CD4 domain has a Tm of about 90 C. 12. The anti-HIV gp120-binding protein of clause 10 or clause 11, wherein the CD4 domain comprises K8C and G99C. 13. The anti-HIV gp120-binding protein of clause 6, comprising any one of SEQ ID NOs: 1-21. 14. The anti-HIV gp120-binding protein of any one of clauses 6 to 13, comprising SEQ ID NO: 11. 15. The anti-HIV gp120-binding protein of any one of clauses 3 to 14, comprising a set of 6 CDRs (CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3) as set forth in any row of Table 1. 16. The anti-HIV gp120-binding protein of clause 15, comprising a CDRH1 of SEQ ID NO: 22, a CDRH2 of SEQ ID NO:23, a CDRH3 of SEQ ID NO:24, a CDRL1 of SEQ ID NO: 25, a CDRL2 of SEQ ID NO:26 and a CDRL3 of SEQ ID NO:27. 17. The anti-HIV gp120-binding protein of any one of the preceding clauses, comprising an immunoglobulin (Ig) scaffold. 18. The anti-HIV gp120-binding protein of clause 15, clause 16 or clause 17, comprising a heavy chain variable (VH) domain and a light chain variable (VL) domain pair as set forth in any row of Table 2. 19. The anti-HIV gp120-binding protein of clause 18, comprising a VH domain of SEQ ID NO: 58 and a VL domain of SEQ ID NO:59 or SEQ ID NO:60. 20. The anti-HIV gp120-binding protein of any one of clauses 17 to 19, comprising an Fc domain. 21. The anti-HIV gp120-binding protein of clause 20, wherein the Fc domain comprises a mutation that increases the half-life of the anti-HIV gp120-binding protein compared to the same anti-HIV gp120-binding protein without said mutation. 22. The anti-HIV gp120-binding protein of clause 21, wherein the Fc domain comprises any one of the following sets of mutations (EU numbering): M428L and N434S (LS); L309D, Q311H and N434S (DHS); M252Y, S254T and T256E (YTE); and H433K and N434F (HN). 23. The anti-HIV gp120-binding protein of clause 22, wherein the Fc domain comprises LS. 24. The anti-HIV gp120-binding protein of any one of the preceding clauses comprising a broadly neutralizing antibody (bNAb). 25. The anti-HIV gp120-binding protein of clause 24, wherein the bNAb is an anti-V3 bNAb. 26. The anti-HIV gp120-binding protein of clause 25, wherein the anti-V3 bNAb comprises a heavy chain (HC) and a light chain (LC) pair as set forth in any row of Table 2. 27. The anti-HIV gp120-binding protein of clause 26, wherein the HC comprises SEQ ID NO: 61 or SEQ ID NO: 62 and the LC comprises SEQ ID NO: 63 or SEQ ID NO:64. 28. The anti-HIV gp120-binding protein of clause 27, wherein the HC comprises SEQ ID NO: 62 and the LC comprises SEQ ID NO:63. 29. The anti-HIV gp120-binding protein of any one of clauses 17 to 19, comprising an anti-V3 bNAb scFv. 30. The anti-HIV gp120-binding protein of clause 29, wherein the scFv comprises a VH domain of SEQ ID NO:58 and a VL domain of SEQ ID NO:59. 31. The anti-HIV gp120-binding protein of clause 29 or clause 30, wherein the C-terminus of the VH domain is fused directly or via a linker to the N-terminus of the VL domain. 32. The anti-HIV gp120-binding protein of clause 29 or clause 30, wherein the C-terminus of the VL domain is fused directly or via a linker to the N-terminus of the VH domain. 33. The anti-HIV gp120-binding protein of clause 31 or clause 32, wherein the linker between the VH domain and the VL domain of the scFv is selected from the group consisting of SEQ ID NOs: 90-95. 34. The anti-HIV gp120-binding protein of clause 33, wherein the linker between the VH domain and the VL domain of the scFv is SEQ ID NO:93. 35. The anti-HIV gp120-binding protein of any one of clauses 29 to 34, wherein the scFv is fused to a human Fc domain directly or via a linker (scFv-Fc). 36. The anti-HIV gp120-binding protein of clause 35, wherein the scFv is fused to a human Fc via a linker selected from the group consisting of SEQ ID NO:90-95. 37. The anti-HIV gp120-binding protein of clause 35, wherein the scFv is fused to a human Fc via a linker of SEQ ID NO:91. 38. The anti-HIV gp120-binding protein of any one of clauses 35 to 37, wherein the Fc domain is as defined in any one of clauses 21 to 23. 39. The anti-HIV gp120-binding protein of any one of clauses 35 to 38, comprising any one of SEQ ID NOs: 152-157. 40. A bispecific anti-HIV gp120-binding protein comprising an anti-V3 bNAb and two copies of a CD4 domain, wherein the C-terminus of one CD4 domain is attached directly or by a linker to the N-terminus of one of the anti-V3 bNAb heavy chains and the C-terminus of the other copy of the CD4 domain is attached directly or by a linker to the N-terminus of the other anti-V3 bNAb heavy chain. 41. The bispecific protein according to clause 40, wherein each CD4 domain is attached via a linker to each of the heavy chains. 42. The bispecific protein according to clause 41, wherein the linker is selected from the group consisting of SEQ ID NOs: 90 to 95. 43. The bispecific protein according to clause 42, wherein the linker is SEQ ID NO:90. 44. The bispecific protein according to any one of clauses 40 to 43, wherein the CD4 domain is selected from the group consisting of SEQ ID NO:1-21. 45. The bispecific protein according to clause 44, wherein the CD4 domain is SEQ ID NO: 11. 46. The bispecific protein according to any one of clauses 40 to 45, wherein the anti-V3 bNAb is selected from the group consisting of bNAb1, bNAb1*, bNAb2, bNAb3, bNAb4, bNAb5 and bNAb6 as set forth in Table 2. 47. The bispecific protein according to clause 46, wherein the anti-V3 bNAb is bNAb1. 48. The bispecific protein according to any one of clauses 40 to 47, wherein the anti-V3 bNAb Fc comprises LS. 49. An anti-HIV gp120-binding protein having two identical heavy chains and two identical light chains, comprising or consisting of: a heavy chain that is at least 95% identical to SEQ ID NO:121 and a light chain that is at least 95% identical to SEQ ID NO:63. 50. An anti-HIV gp120-binding protein consisting of two heavy chains of SEQ ID NO: 121 and two light chains of SEQ ID NO:63. 51. An anti-HIV gp120-binding protein comprising or consisting of a sequence that is at least 95% identical to any one of SEQ ID NOs: 152-157. 52. An anti-HIV gp120-binding protein consisting of SEQ ID NO:155. 53. A pharmaceutical composition comprising the anti-HIV gp120-binding protein as defined in any one of the preceding clauses and a pharmaceutically acceptable excipient. 54. A method of treating or preventing an HIV infection in a human comprising administering to the human an anti-HIV gp120-binding protein according to any one of clauses 1 to 52, or a pharmaceutical composition according to clause 53, whereby viral load in the human is decreased. 55. An anti-HIV gp120-binding protein according to any one of clauses 1 to 52, or a pharmaceutical composition according to clause 53, for use in treating or preventing an HIV infection in a human. 56. Use of an anti-HIV gp120-binding protein according to any one of clauses 1 to 52, or a pharmaceutical composition according to clause 53, in the manufacture of a medicament for treating or preventing an HIV infection in a human. 57. A kit comprising in separate containers: an anti-HIV gp120-binding protein according to any one of clauses 1 to 52 and an anti-viral drug that inhibits cellular entry, replication, or transcription of HIV in a human. 58. The kit according to clause 57, wherein the antiviral drug is selected from the group consisting of: Nucleoside Reverse Transcriptase Inhibitors (NRTIs), Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs), Protease Inhibitors (PIs), Entry Inhibitors, Integrase Strand Transfer Inhibitors (INSTI), Maturation Inhibitors (MIs), Capsid Inhibitors (CIs) and Nucleoside Reverse Transcriptase Translocation Inhibitors (NRTTIs) 59. The kit according to clause 58, wherein the antiviral drug is an INSTI. 60. The kit according to clause 59, wherein the INSTI is dolutegravir or cabotegravir. 61. A nucleic acid sequence that encodes an anti-HIV gp120-binding protein according to any one of clauses 1 to 52. 62. An expression vector that comprises the nucleic acid sequence of clause 61. 63. A host cell that comprises the nucleic acid sequence of clause 61 or the expression vector of clause 62. 64. A host cell that comprises two expression vectors: a first expression vector comprising a nucleic acid sequence encoding a heavy chain of SEQ ID NO:121; and a second expression vector comprising a nucleic acid sequence encoding a light chain of SEQ ID NO:63. 65. A method of producing an anti-HIV gp120-binding protein, comprising culturing the host cell as defined in clauses 63 or 64 under conditions suitable for expression of said nucleic acid sequence or vector, whereby an anti-HIV gp120-binding protein is produced. 66. A soluble CD4 domain having a Tm above 70 C. 67. A soluble CD4 domain comprising one or more stabilizing mutations selected from the group consisting of K8C, K8I, K8V, T11C, E13C, K21C, Q25E, H27C, H27D, G38C, N52W, R58N, R58T, R58V, L61M, G65C, I70C, K72C, E87G, E91H, E91Q, and G99C. 68. The soluble CD4 domain of clause 66 or clause 67 having a Tm of between 70 C. and 95 C. 69. The soluble CD4 domain of clause 68 having a Tm of about 90 C. 70. The soluble CD4 domain of any one of clauses 66 to 69, comprising K8C and G99C. 71. The soluble CD4 domain of any one of clauses 66 to 68, comprising K8I. 72. The soluble CD4 domain of any one of clauses 66 to 68, comprising K8V. 73. The soluble CD4 domain of any one of clauses 66 to 68, comprising T11C and K72C 74. The soluble CD4 domain of any one of clauses 66 to 68, comprising any one of SEQ ID NOs: 5-21. 75. The soluble CD4 domain of clause 74, comprising SEQ ID NO:11. 76. The soluble CD4 domain of any one of clauses 66 to 75, wherein the CD4 domain is fused directly or via a linker to a human Fc domain. 77. The soluble CD4 domain of clause 76, wherein the Fc domain comprises LS. 78. The soluble CD4 domain of clause 76 or clause 77, wherein the linker is selected from the group consisting of SEQ ID NOs: 90-95.
EXAMPLES
Example 1Antigen Binding Protein Production
(227) Plasmids encoding the antigen binding proteins of the invention were expressed in EXPI293 or FREESTYLE 293-F cells using the manufacturer's standard protocol (ThermoFisher Scientific, Waltham, MA). The expressed medium was harvested by centrifugation (4000 rpm for 10 min) and the antigen binding proteins were purified by filtration through a 0.22 m filter (Millipore Sigma, Burlington, MA) and fast protein liquid chromatography (FPLC) (KTATM Pure, Cytiva, Marlborough MA). The medium was then passed through a Mabselect SuRe column (Cytiva, Marlborough MA) to capture the antigen binding proteins and the column was washed sequentially with phosphate-buffered saline (PBS) before elution.
(228) The antigen binding proteins were then exchanged into a final buffer by using dialysis, a desalting column and preparative size exclusion column (SEC). The purity of the antigen binding proteins was evaluated by using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and on a size exclusion column on a high-performance liquid chromatography system (SEC-HPLC).
(229) Antigen binding protein concentrations were determined by measuring absorbance at 280 nm wavelength (A280) on a NanoDrop machine (ThermoFisher Scientific, Waltham, MA), and their molecular mass was measured by using liquid chromatography-mass spectrometry (LC-MS) to confirm their identity.
(230) The endotoxin level in the final purified products was measured on an ENDOSAFE system (Charles River Labs, Wilmington MA) to make sure it was sufficiently low (usually <1 EU (Endotoxin Unit)/mg of protein) for downstream anti-viral studies.
Example 2Anti-Viral Activity
(231) The anti-viral activity of the antigen binding proteins was measured in a pseudotyped virus (PSV) assay. Pseudotyped HIV-1 virus (PSV) contains deletions in the genome that make it unable to produce infectious virions, but it can be used to measure the activity of cell entry inhibitors (i.e., molecules that prevent the binding of HIV-1 virions to the target cell membrane and/or prevent entry of HIV-1 into target cells), which include the antigen binding proteins of the invention.
(232) PSV was produced in HEK-293T cells (ATCC, Manassas VA) by co-transfecting expression plasmids encoding the HIV-1 gp160 envelope gene and an HIV-1 backbone plasmid using TRANSIT-2020 transfection reagent (Mirus Bio, Madison WI). A panel of HIV-1 PSVs expressing different gp160 envelope trimers was generated to evaluate the effectiveness of the antigen binding proteins of the invention against a wide spectrum of HIV-1 strains.
(233) i. ACTOne Cells
(234) The genome of PSV used in this assay contains a luciferase gene that is expressed once the virus enters cells. Accordingly, the luminescence signal (after adding a substrate of luciferase) can be used to determine the level of viral infection.
(235) The 50% tissue culture infectious dose (TCID) of a single thawed aliquot of each batch of PSV was determined in ACTOne cells. The ACTOne cell-line used in this assay was derived in-house from a genetically engineered 293T cell clone that expresses CD4, CXCR4, and CCR5. Cells were maintained in growth medium composed of Dulbecco's modified Eagle's medium (DMEM, Life Technologies) at 37 C. in a humidified 5% CO.sub.2-95% air environment. Cell monolayers were split by treatment with Trypsin-EDTA (0.05%).
(236) To run the anti-viral assay, ACTOne cells were detached by treating the cell culture flask with trypsin (trypsinization) and resuspended in growth medium containing 2% of DMSO to a density of 2.510.sup.5 cells/ml. One hundred l of such cells was added to 10 l of antigen binding protein pre-loaded in a 96-well plate. Ninety l of PSV was then added to each well. The assay plates were incubated at 37 C. in a humidified incubator at 5% CO.sub.2 level. Plates were developed after 72 hours of incubation by adding 50 l of BRIGHTGLO luciferase reagent (Promega, Madison WI) to each well, and transferring the plates to an ENVISION multilabel plate reader (PerkinElmer, Waltham MA) to measure the luminescence and determine the level of virus that had infected the cells. The higher the luminescence signal, the higher the level of infection.
(237) Raw data were analyzed using an in-house template in an IDBS system to calculate half-maximal inhibitory concentration (IC50) values which reflects the activity of the antigen binding proteins of the invention at inhibiting viral entry (the smaller the number is, the more active the molecule is).
(238) ii. TZM.bl Cells
(239) Alternatively, the PSV assay was carried out using a luciferase-based assay in a TZM.bl cell line. The TZM-bl cell line is derived from a Hela cell clone that was engineered to express CD4, CCR5 and CXCR4 and to contain integrated reporter genes for firefly luciferase and E. coli -galactosidase under the control of an HIV-1 long terminal repeat (Wei et al., Antimicrobial agents and chemotherapy 46:1896-905 (2002)) permitting sensitive and accurate measurements of infection.
(240) The detailed materials and methodology have been described elsewhere (Mentefiori, Curr. Protoc. Immunol., 2005, Chapter 12; Seaman et al., Journal of Virology, February 2010, 84 (3), p. 1439-1452). In brief, the assay measures the reduction in luciferase reporter gene expression in TZM.bl cells following a single round of virus infection.
(241) Five-fold serial dilutions of the antigen binding proteins of the invention, from 50 g/ml to 3.2 ng/ml, were performed in duplicate in 10% DMEM growth medium (100 u/well). An amount of 200 TCID50 (50% tissue culture infectious dose) of virus was added to each well in a volume of 50 l, and the plates were incubated for 1 h at 37 C.
(242) TZM.bl cells were then added (110.sup.4/well in a 100-l volume) in 10% D-MEM growth medium containing DEAE-dextran (Sigma, St. Louis, MO) at a final concentration of 11 g/ml. Assay controls included TZM.bl cells alone (cell control) and TZM.bl cells with virus (virus control).
(243) Following a 48-hour incubation at 37 C., 150 l of assay medium was removed from each well and 100 l of BRIGHTGLO luciferase reagent (Promega, Madison, WI) was added. The cells were allowed to lyse for 2 min, and then 150 l of the cell lysate was transferred to a 96-well black solid plate, and luminescence was measured using a Victor 3 luminometer (Perkin Elmer).
(244) The 50% and 80% inhibitory concentration (IC50 and IC80) values were calculated as the serum dilution that caused a 50% and 80% reduction respectively, in relative luminescence units (RLU) compared to the level in the virus control wells after subtraction of cell control RLU. All data were analyzed with 5-parameter curve fitting using neutralizing antibody analysis software provided by the CAVD Vaccine Immunology Statistical Center.
Example 3Stability of Soluble CD4 Domains
(245) All soluble human CD4 domains tested contain a set of base mutations in human CD4 domain 1 (D1) over the wild-type sequence (SEQ ID NO:3) that enable the folding of human CD4 D1 on its own. Soluble CD4 D1 with this set of mutations is known as mD1.22 (Chen et al., J Virol. 2014 January; 88 (2): 1125-39) and the mutations therein consist of: L5Y, S23N, A55V, 176P, L96V, and F98V (SEQ ID NO:4, also referred to as D1m herein).
(246) To achieve better developability and pharmacokinetics, further mutations were introduced into mD1.22 (SEQ ID NO:4) to enhance its thermal stability. The additional stabilizing mutations were designed based on several methodologies: 1) computational simulation by using Free Energy Perturbation (FEP+, Schrodinger, New York, NY USA); 2) computational simulation by using disulfide-bond scan in Molecular Operating Environment program (MOE, Chemical Computing Group, Montreal Canada); and 3) panning a library of human CD4 D1 with each residue mutated, one by one, to the other 19 types of amino acids (site saturation mutagenesis, TWIST BioScience, San Francisco, CA USA) using phage display under thermally challenging conditions (i.e., incubating the phage at room temperature, 70 C., and 80 C., then selecting the CD4 domain variants that can still bind to recombinant HIV-1 gp120 (CN54 strain, Acro Biosystems, Beijing China)).
(247) The best performing variants (SEQ ID NOs: 5-21) were fused with 6His tag at their C-termini, expressed and purified from mammalian cells using methods as described in Example 1, except that purification was via a Ni-NTA resin (Cytiva, Marlborough MA) instead of Mabselect SuRe column, with standard protocol from the vendor.
(248) These purified CD4 D1 variants (with C-terminal 6xHis tag) were then evaluated to determine their melting temperature (Tm, using Prometheus System, NanoTemper, Mnchen Germany), which indicates thermal stability, as well as their anti-viral activity against HIV-1 pseudotyped virus (see Example 2 above for methods using ACTOne cells).
(249) As shown in Table 3 below and in
(250) TABLE-US-00006 TABLE 3 Melting temperature of soluble CD4 domains CD4 Domain SEQ ID NO Tm ( C.) D1m 4 68.2 D1m-E91Q 6 72.4 D1m-E91H 7 72.6 D1m-E87G 8 71.9 D1m-N52W 9 72 D1m-K8V 5 77.1 D1m-K8I 10 75.9 D1m-K8C-G99C 11 88.8 D1m-T11C-K72C 12 79.9 D1m-E13C-I70C 13 78.8 D1m-H27C-G38C 14 90.1 D1m-K21C-G65C 15 80.1
Example 4Antigen Binding Protein Format and Linker Length
(251) The fusion position of the CD4 domain in the anti-V3 bNAbs (e.g., whether to fuse the CD4 domain to the light chain or heavy chain or both, whether to fuse the CD4 domain to the N-terminus or C-terminus of these chains, or whether to fuse the CD4 domain in the middle of the heavy chain (in between CH1 and CH2 domains)) has an effect on the anti-viral potency of the resulting bispecific as shown in Table 4.1 and Table 5 below.
(252) We observed that the most potent bispecific molecule resulted from fusing CD4 D1 to the N-terminus of the heavy chain of bNAb1 (molecule 1 in Table 4.1, which neutralized 6 envelopes with IC50<160 pM and 1 envelope with IC50 about 3 nM in PSV assay). In this bispecific format, the linker length between the CD4 domain and bNAb1 heavy chain N-terminus does not particularly affect anti-viral activity (
(253) As shown in
(254) TABLE-US-00007 TABLE 4.1 (see also FIG. 7) IC50 (nM) of different bNAb1-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in a PSV assay (ACTOne cells) bNAb1-Derived Molecules Envelope 1 2 3 4 5 6 7 8 9 10 11 42368 0.16 0.91 1.49 4.90 1.24 15.73 5.60 1.44 4.32 1.58 >500 CC1/85 0.04 0.05 0.49 0.01 0.01 0.03 0.04 0.07 7.02 1.98 0.03 NL4-3 0.05 0.17 0.17 0.10 0.14 0.52 0.45 0.22 0.31 0.31 131.20 T278.50 0.12 0.12 1.29 0.04 0.02 0.06 0.13 0.11 3.99 1.43 0.17 X2088.C9 0.07 0.86 0.72 13.28 1.50 67.68 18.25 5.25 17.49 9.20 402.35 ZM106.9 0.05 0.18 0.38 0.04 0.03 0.05 0.07 0.09 26.02 15.14 0.07 CAP45.2.00.G3 3.73 2.68 2.53 13.89 2.13 20.44 102.16 20.41 131.43 20.48 >500 Table 4.1 Molecule Key: 1 = D1m-K8C-G99C_1xG4S_bNAb1 (SEQ ID NOs: 121 + 63) 2 = D1m-K8C-G99C_1xG4S_bNAb1-LC (SEQ ID NOs: 62 + 115) 3 = D1m-1xG4S_bNAb1-BothChains (SEQ ID NOs: 102 + 109) 4 = bNAb1-mid_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 116 + 63) 5 = bNAb1-HC_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 117 + 63) 6 = bNAb1-LC_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 62 + 118) 7 = D1m_His (SEQ ID NO: 4*) + bNAb1 (SEQ ID NOS: 62 + 63) (combo) 8 = D1m-K8C-G99C_Fc (SEQ ID NO: 158) + bNAb1 (SEQ ID NOs: 62 + 63) (combo) 9 = D1m_His (SEQ ID NO: 4*) 10 = D1m-K8C-G99C_Fc (SEQ ID NO: 158) 11 = bNAb1 (SEQ ID NOS: 62 + 63) *plus a 6xHis tag (six C-terminal histidine residues)
(255) TABLE-US-00008 TABLE 4.2 IC50 (nM) of a bNAb1-derived bispecific against entry inhibitor resistant envelopes in a PSV assay (ACTOne cells) bNAb1-derived Type Envelope bispecific 10E8-insensitive KER2008.12 0.05 T266-60 0.06 ZM106.9 0.05 X2088.c9 0.07 MB539.2B7 0.04 JR2 with W680R 0.03 and K683Q N6-insensitive BL01 0.07 T278-50 0.12 6471_V1_C16 0.06 CH0219_E4 0.08 Temsavir- and Ibalizumab- 21-116102 0.36 insensitive 21-116108 0.01 Maraviroc-insensitive MP5.7 0.13 MP11.38 0.05 MP35.2 0.13 MP49.20 0.02 MP53.36 0.04 Table 4.2 Molecule Key: 1 = D1m-K8C-G99C_1xG4S_bNAb1 (SEQ ID NOs: 121 + 63)
(256) TABLE-US-00009 TABLE 5 IC50 (nM) of different bNAb2-, bNAb3- and bNAb4-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb3-Derived bNAb2-Derived Molecules Molecules Envelope 1 2 3 4 5 6 7 8 9 10 42368 1.79 0.75 1.24 269.20 4.59 2.03 >500 0.37 0.17 5.01 CC1/85 0.64 0.46 0.33 107.46 9.44 1.98 >500 0.29 0.07 4.07 NL4-3 0.44 0.26 0.42 8.44 0.80 0.22 150.32 0.08 0.07 0.11 T278.50 0.16 0.10 0.09 0.48 0.30 0.14 0.43 0.02 0.03 0.02 X2088.C9 0.06 0.05 0.05 0.07 0.02 0.09 0.09 0.06 0.03 0.05 ZM106.9 0.05 0.02 0.02 0.04 0.01 0.02 0.03 0.03 0.03 0.01 CAP45.2.00.G3 0.10 0.10 0.11 2.49 12.51 3.63 36.53 1.69 0.25 96.75 HIV-2-AID 14.16 19.13 20.28 136.88 >500 * >500 * * * HIV-2-ATM88 1.34 1.22 2.28 2.06 70.58 * >500 * * * HIV-2-HCC-01 35.94 17.47 24.36 >500 >500 * >500 * * * bNAb3-Derived Molecules bNAb4-Derived Molecules CD4 Controls Envelope 11 12 13 14 15 16 17 18 42368 2.12 >500 0.50 8.78 3.42 >500 4.32 1.58 CC1/85 1.15 38.33 0.10 1.39 0.77 2.36 7.02 1.98 NL4-3 0.06 0.15 0.19 0.86 0.48 327.61 0.31 0.31 T278.50 0.02 0.01 0.13 0.23 0.41 0.47 3.99 1.43 X2088.C9 0.03 0.03 0.03 0.01 0.04 0.02 17.49 9.20 ZM106.9 0.03 0.03 0.03 0.02 0.03 0.03 26.02 15.14 CAP45.2.00.G3 61.81 >500 13.18 469.16 42.98 >500 131.43 20.48 HIV-2-AID * * * * * * 103.54 24.35 HIV-2-ATM88 * * * * * * 3.93 2.72 HIV-2-HCC-01 * * * * * * >500 >300 * not tested Table 5 Molecule Key: 1 = D1m-K8C-G99C_2xG4S_bNAb2 (SEQ ID NOs: 137 + 69) 2 = D1m-K8C-G99C_3xG4S_bNAb2 (SEQ ID NOs: 138 + 69) 3 = D1m-K8C-G99C_4xG4S_bNAb2 (SEQ ID NOs: 139 + 69) 4 = bNAb2-LC_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 68 + 143) 5 = bNAb2 (SEQ ID NOs: 68 + 69) + D1m_His (SEQ ID NO: 4*) (combo) 6 = bNAb2 (SEQ ID NOs: 68 + 69) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 7 = bNAb2 (SEQ ID NOs: 68 + 69) 8 = D1m-K8C-G99C_1xG4S_bNAb3 (SEQ ID NOs: 144 + 74) 9 = D1m-K8C-G99C_4xG4S_bNAb3 (SEQ ID NOs: 145 + 74) 10 = bNAb3 (SEQ ID NOs: 73 + 74) + D1m_His (SEQ ID NO: 4*) (combo) 11 = bNAb3 (SEQ ID NOs: 73 + 74) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 12 = bNAb3 (SEQ ID NOs: 73 + 74) 13 = D1m-K8C-G99C_4xG4S_bNAb4 (SEQ ID NOs: 147 + 79) 14 = bNAb4 (SEQ ID NOs: 78 + 79) + D1m_His (SEQ ID NO: 4*) (combo) 15 = bNAb4 (SEQ ID NOs: 78 + 79) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 16 = bNAb4 (SEQ ID NOs: 78 + 79) 17 = D1m_His (SEQ ID NO: 4*) 18 = D1m-K8C-G99C_Fc (SEQ ID NO: 158) *plus a 6xHis tag (six C-terminal histidine residues)
(257) Thermal stabilization of CD4 D1 (see Example 3 above) further enhanced the PK of the bispecific molecules (D1m-KBC-G99C_1xG4S_bNAb1, SEQ ID NOs: 121 and 63; D1m-T11C-K72C_1xG4S_bNAb1, SEQ ID NOs: 122 and 63; D1m-K8I_1xGAS_bNAb1, SEQ ID NOs: 119 and 63; and D1m-KBV_1xGAS_bNAb1, SEQ ID NOs: 120 and 63) as shown in Table 6 below.
(258) TABLE-US-00010 TABLE 6 The effect of (1) linker length between the CD4 domain and bNAb1 heavy chain, and (2) thermal stability of CD4 D1, on the PK of bispecific molecules in hFcRn mice (Tg32) Tm of Molecule CD4 D1 Half-life Clearance (SEQ ID NOs) ( C.) (days) (ml/day/kg) bNAb1 (62 + 63) 5.8 21.6 D1m_4xG4S_bNAb1 (105 + 63) 68.2 2.2 144 D1m_1xG4S_bNAb1 (102 + 63) 68.2 4.1 31.7 D1m-K8C-G99C_1xG4S_bNAb1 88.8 10.2 7.4 (121 + 63) D1m-T11C- 79.9 8.9 12.9 K72C_1xG4S_bNAb1 (122 + 63) D1m-K8I_1xG4S_bNAb1 75.9 9.3 12 (119 + 63) D1m-K8V_1xG4S_bNAb1 77.1 7.3 12.8 (120 + 63)
(259) Accordingly, the best molecules for further development contain shorter linker lengths between the CD4 domain and bNAb (1xG4S) and contain a thermally stable CD4 domain(s).
Example 5Anti-Viral Activity of bNAb1-derived Bispecific Molecules
(260) As shown in
(261) Interestingly, soluble CD4 has been considered to have negative synergy with bNAb1 (Ivan et al., Plos Biol. 17 (1), January 2019), based on the result that mixing soluble CD4 with bNAb1 can weaken its anti-viral activity. We found that when soluble CD4 is fused with bNAb1 instead of being mixed, the anti-viral activity is enhanced dramatically.
(262)
(263) As shown in Table 4.1 above, although all of the bispecific molecules tested are able to inhibit viral entry, the most consistent and potent bispecific molecule is when the CD4 domain is fused to the N-termini of each of the heavy chains of bNAb1, which correlates well with the structure-based design (
(264) Furthermore, as shown in Table 4.2 above, when tested in PSV assay against the HIV-1 envelopes insensitive to several entry inhibitors (10E8, N6, Temsavir, Ibalizumab, and Maraviroc), the bispecific molecule (SEQ ID NOs: 121 and 63) fully neutralized all these envelopes with IC50s <400 pM.
(265) In addition, the most potent bispecific format (i.e. fusing CD4 D1 to the N-terminus of bNAb1 heavy chain) was converted to a single open-reading-frame (ORF) version, by replacing the Fab arm with a scFv fragment of bNAb1. As shown in
(266) Given that these single-ORF molecules are each encoded by a single <2 kb gene and contain an Fc domain for an increased half-life, they can be readily delivered by gene therapy vehicles, such as adeno-associated virus (AAV), enabling them to be constantly secreted into circulation at a therapeutic concentration. Such a strategy would result in an ultra-long acting therapy against HIV-1.
Example 6Anti-Viral Activity of bNAb6-Derived Bispecific Molecules
(267)
(268) The plot of IC50 values (
(269) Table 7 shows the anti-viral activity of the bispecific molecules (D1m_4xG4S_bNAb6, SEQ ID NOs: 151 and SEQ ID NOs: 89; D1mD2_4xG4S_bNAb6, SEQ ID NO:150 and SEQ ID NO:89; and D1m-K8C-G99C_1xG4S_bNAb6 (SEQ ID NOs: 362+89)) and control molecules against HIV-1 strains resistant to bNAb6 antibody. As can be seen, the bNAb6-derived bispecific molecules are much more potent than the mixtures against double-resistant or insensitive strains.
(270) TABLE-US-00011 TABLE 7 IC50 (nM) of bNAb6-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb6-Derived Molecules Envelope 1 2 3 4 5 6 7 CAP45.2.00.G3 17.79 >500 5.90 >500 131.43 >500 >500 CH119 8.30 93.62 2.60 49.76 90.66 119.94 >500 BJOX2000 17.27 45.23 3.10 16.96 70.87 33.40 >500 X1632_S2_B10 1.38 20.26 0.75 19.70 16.25 14.94 >500 TRO11 1.99 16.55 1.14 14.64 68.43 17.27 >500 T278.50 0.71 9.26 1.08 9.51 3.99 16.02 >500 LAI 1.52 6.76 0.93 7.46 8.79 5.41 >500 42368 1.30 6.05 0.54 5.75 4.32 7.29 >500 NL4-3 0.17 1.31 0.11 0.75 0.31 0.74 >500 HXB2 0.06 0.08 0.03 0.06 0.11 0.07 >500 bNAb6- Derived Molecules Envelope 8 9 7 10 X2088.C9 0.17 0.15 0.15 9.20 ZM106.9 0.10 0.10 0.08 15.14 3637_V5_C3 103.00 328.50 >500 >500 3468_V1_C12 20.85 47.70 >500 64.15 Q461_E2 4.00 12.70 >500 11.76 3326_V4_C3 72.10 483.00 >500 >500 42368 0.92 2.13 >500 1.58 NL4-3 0.27 * >500 0.31 T278.50 1.33 * >500 1.43 CAP45.2.00.G3 9.64 * >500 20.48 CC1/85 0.42 * * 1.98 HIV-2-HCC-01 175.65 * * >300 * not tested Table 7 Molecule Key: 1 = D1m_4xG4S_bNAb6 (SEQ ID NOs: 151 + 89) 2 = D1m_His (SEQ ID NO: 4*) + bNAb6 (SEQ ID NOs: 88 + 89) 3 = D1mD2_4xG4S_bNAb6 (SEQ ID NOs: 150 + 89) 4 = D1mD2_His (SEQ ID NO: 2*) + bNAb6 (SEQ ID NOs: 88 + 89) 5 = D1m_His (SEQ ID NO: 4*) 6 = D1mD2_His (SEQ ID NO: 2*) 7 = bNAb6 (SEQ ID NOs: 88 + 89) 8 = D1m-K8C-G99C_1xG4S_bNAb6 (SEQ ID NOs: 362 + 89) 9 = bNAb6 (SEQ ID NOs: 88 + 89) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 10 = D1m-K8C-G99C_Fc (SEQ ID NO: 158) *plus a 6xHis tag (six C-terminal histidine residues)
Example 7Anti-Viral Activity of Further Anti-V3 bNAb-Derived Bispecific Molecules
(271) Additional bispecific molecules comprising CD4 domains fused to other anti-V3 bNAbs (bNAb2, bNAb3, bNAb4,) were tested, with the results shown in Table 5 above and Tables 8-23 below, a similar synergistic anti-viral activity was observed against one or more envelopes.
(272) Accordingly, the strategy of fusing a soluble CD4 domain to anti-V3 loop bNAb can be generally applied to enhance the potency and spectrum of these bNAbs.
Conclusions
(273) On the free HIV-1 virus, the V3 loop of gp120 is in its native closed state. The V3 loop is known to adopt various conformations (from different structures in Protein Databank), indicating its flexibility. During HIV-1 infection, the binding of gp120 to cell surface CD4 triggers conformational changes of the V3 loop to open itself to bind co-receptors such as CXCR4 or CCR5.
(274) Anti-V3 bNAbs mainly recognizes a pattern of glycans on the V3 loop of gp120, along with the backbone atoms of a few amino acid residues in the V3 loop (Krumm et al., Retrovirology 13 (8), 2016). Such plasticity of the V3 loop may facilitate the binding of anti-V3 loop bNAbs to this loop when it is opened by CD4 binding.
(275) We hypothesize that when soluble CD4 and anti-V3 bNAbs are simply mixed together, the conformational change of the V3 loop triggered by soluble CD4 may be too transient for the anti-V3 bNAb to capture, therefore no synergistic activity is observed. But, in the context of bispecific molecules, when CD4 binds to the CD4 binding site (CD4bs) on gp120, the anti-V3 bNAb is at such high local concentration that it can immediately capture the exposed V3 loop glycans; this in turn could stabilize the binding of soluble CD4 to gp120 and form a positive feedback loop.
(276) TABLE-US-00012 TABLE 8 IC50 (nM) of different bNAb5-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb5-Derived Molecules Controls Envelope Bispecific Combo mAb CD4 42368 1.11 2.35 >500 1.58 CC1/85 0.67 2.53 >500 1.98 NL4-3 0.26 0.26 2.62 0.31 T278.50 3.21 1.59 96.06 1.43 X2088.C9 0.54 0.14 0.17 9.20 ZM106.9 0.94 1.09 2.04 15.14 CAP45.2.00.G3 44.41 63.82 >500 20.48 25710 0.51 0.49 8.10 1.32 TRO11 0.91 0.58 0.39 146.64 398F1 4.85 1.35 1.83 90.47 CNE8 134.25 240.63 51.37 346.64 X2278 0.60 5.46 9.53 44.56 BJOX2000 1.11 1.86 126.82 2.34 X1632 2.32 2.38 >500 1.82 CE1176 1.04 2.59 57.07 4.22 246F3 7.18 5.27 40.90 4.28 CH119 2.63 3.22 63.54 3.25 CE0217 2.42 2.19 409.83 2.53 CNE55 20.92 36.59 3300.62 41.82 JR-CSF 0.31 0.45 0.64 5.47 JRFL 0.42 1.16 58.23 1.12 Table 8 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb5 (SEQ ID NOs: 148 + 84) Combo = bNAb5 (SEQ ID NOs: 83 + 84) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb5 (SEQ ID NOs: 83 + 84) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(277) TABLE-US-00013 TABLE 9 IC50 (nM) of different bNAb7-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb7-Derived Molecules Controls Envelope Bispecific Combo mAb CD4 42368 1.38 2.40 >500 1.58 X2088.C9 0.09 0.19 0.22 9.20 ZM106.9 0.03 0.03 0.04 15.14 3326_V4_C3 17.63 >500 >500 >500 3637_V5_C3 260.71 388.10 >500 >500 3468_V1_C12 2.50 32.37 >500 64.15 620345_C1 4.03 5.83 >500 99.28 0260.v5.c36 0.16 0.22 0.75 119.29 TH976_17 70.02 35.04 >500 39.05 Q461_E2 14.61 20.92 >500 11.76 928_28 0.43 0.57 32.73 0.77 YU-2 0.07 0.16 0.58 0.20 6471_V1_C16 0.59 0.50 >500 5.07 93UG065 50.99 40.15 >500 142.61 CC1/85 0.21 * * 1.98 NL4-3 0.33 * * 0.31 T278.50 1.79 * * 1.43 CAP45.2.00.G3 0.22 * * 20.48 HIV-2-HCC-01 183.57 * * >300 * not tested Table 9 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb7 (SEQ ID NO: 338 + 259) Combo = bNAb7 (SEQ ID NOs: 258 + 259) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb7 (SEQ ID NOs: 258 + 259) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(278) TABLE-US-00014 TABLE 10 IC50 (nM) of different bNAb8-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb8-Derived Molecules Controls Envelope Bispecific Combo mAb CD4 X2088.C9 0.11 0.22 0.23 9.20 ZM106.9 0.02 0.03 0.03 15.14 3637_V5_C3 104.00 305.50 >500 >500 3468_V1_C12 0.06 0.81 1.70 64.15 Q461_E2 7.06 6.59 >500 11.76 3326_V4_C3 75.70 367.00 >500 >500 42368 0.97 1.35 >500 1.58 CC1/85 0.14 * * 1.98 NL4-3 0.14 * * 0.31 T278.50 1.20 * * 1.43 CAP45.2.00.G3 2.72 * * 20.48 HIV-2-HCC-01 33.29 * * >300 * not tested Table 10 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb8 (SEQ ID Nos: 339 + 264) Combo = bNAb8 (SEQ ID Nos: 263 + 264) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb8 (SEQ ID Nos: 263 + 264) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(279) TABLE-US-00015 TABLE 11 IC50 (nM) of different bNAb9-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb9-Derived Molecules Controls Envelope Bispecific Combo mAb CD4 42368 0.15 1.35 >500 1.58 X2088.C9 0.18 9.50 >500 9.20 ZM106.9 0.08 16.28 >500 15.14 3326_V4_C3 3.87 >500 >500 >500 3637_V5_C3 6.74 >500 >500 >500 3468_V1_C12 1.51 23.22 >500 64.15 620345_C1 1.02 3.25 >500 99.28 0260.v5.c36 0.19 0.14 0.26 119.29 TH976_17 20.42 27.24 >500 39.05 Q461_E2 0.55 18.54 >500 11.76 928_28 0.20 0.42 >500 0.77 YU-2 0.12 0.09 0.36 0.20 6471_V1_C16 0.11 1.17 >500 5.07 93UG065 0.18 31.44 >500 142.61 CC1/85 0.03 * * 1.98 NL4-3 0.07 * * 0.31 T278.50 0.14 * * 1.43 CAP45.2.00.G3 1.03 * * 20.48 HIV-2-HCC-01 1.68 * * >300 * not tested Table 11 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb9 (SEQ ID NOs: 340 + 269) Combo = bNAb9 (SEQ ID NOs: 268 + 269) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb9 (SEQ ID NOs: 268 + 269) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(280) TABLE-US-00016 TABLE 12 IC50 (nM) of different bNAb10-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb10-Derived Molecules Controls Envelope Bispecific Combo mAb CD4 42368 0.17 2.29 189.62 1.58 X2088.C9 0.17 9.94 233.05 9.20 ZM106.9 0.09 24.96 71.31 15.14 3326_V4_C3 14.28 >500 >500 >500 3637_V5_C3 0.36 439.54 >500 >500 3468_V1_C12 0.21 38.64 258.28 64.15 620345_C1 0.33 0.85 >500 99.28 0260.v5.c36 0.17 0.29 0.48 119.29 TH976_17 18.49 31.63 >500 39.05 Q461_E2 0.41 15.03 >500 11.76 928_28 0.17 0.42 320.41 0.77 YU-2 0.12 0.23 0.81 0.20 6471_V1_C16 0.12 1.23 91.84 5.07 93UG065 0.13 0.12 0.21 142.61 CC1/85 0.04 * * 1.98 NL4-3 0.08 * * 0.31 T278.50 0.16 * * 1.43 CAP45.2.00.G3 4.55 * * 20.48 HIV-2-HCC-01 0.95 * * >300 * not tested Table 12 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb10 (SEQ ID Nos: 341 + 274) Combo = bNAb10 (SEQ ID Nos: 273 + 274) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb10 (SEQ ID Nos: 273 + 274) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(281) TABLE-US-00017 TABLE 13 IC50 (nM) of different bNAb11-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb11-Derived Molecules Controls Envelope Bispecific Combo mAb CD4 42368 0.48 2.65 >500 1.58 X2088.C9 0.15 7.06 95.12 9.20 ZM106.9 0.07 14.74 73.68 15.14 3326_V4_C3 51.71 146.99 >500 >500 3637_V5_C3 6.73 410.00 >500 >500 3468_V1_C12 1.80 25.99 >500 64.15 620345_C1 1.27 10.97 438.41 99.28 0260.v5.c36 0.20 0.42 0.57 119.29 TH976_17 17.37 55.43 >500 39.05 Q461_E2 0.51 12.32 >500 11.76 928_28 0.27 0.46 >500 0.77 YU-2 0.10 0.26 0.76 0.20 6471_V1_C16 0.25 0.71 253.40 5.07 93UG065 0.22 24.72 89.50 142.61 CC1/85 0.09 * * 1.98 NL4-3 0.16 * * 0.31 T278.50 0.26 * * 1.43 CAP45.2.00.G3 5.82 * * 20.48 HIV-2-HCC-01 8.69 * * >300 * not tested Table 13 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb11 (SEQ ID NOs: 342 + 279) Combo = bNAb11 (SEQ ID NOs: 278 + 279) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb11 (SEQ ID NOs: 278 + 279) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(282) TABLE-US-00018 TABLE 14 IC50 (nM) of different bNAb12-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb12-Derived Molecules Controls Envelope Bispecific1 Bispecific2 Bispecific3 Bispecific4 Combo mAb CD4 X2088.C9 0.59 13.09 4.43 >500 7.37 >500 9.20 ZM106.9 0.13 0.13 0.27 3.37 1.55 1.85 15.14 3637_V5_C3 9.45 130.00 62.45 >500 474.07 >500 >500 3468_V1_C12 0.82 2.94 4.41 103.85 16.30 343.72 64.15 Q461_E2 0.26 0.3 0.5 4.5 5.16 41.90 11.76 3326_V4_C3 9.63 63.69 81.80 >500 266.93 >500 >500 42368 0.15 6.2 1.3 167.0 1.84 344.45 1.58 620345_C1 0.33 * * * 2.13 179.12 99.28 0260.v5.c36 0.44 * * * 0.50 5.19 119.29 TH976_17 29.83 * * * 36.40 >500 39.05 928_28 0.46 * * * 0.48 482.30 0.77 YU-2 0.21 * * * 0.29 77.41 0.20 6471_V1_C16 0.26 * * * 0.59 >500 5.07 93UG065 0.42 * * * 0.86 3.17 142.61 CC1/85 0.12 * * * * * 1.98 NL4-3 0.14 * * * * * 0.31 T278.50 0.19 * * * * * 1.43 CAP45.2.00.G3 1.29 * * * * * 20.48 HIV-2-HCC-01 4.04 * * * * * >300 * not tested Table 14 Molecule Key: Bispecific1 = D1m-K8C-G99C_1xG4S_bNAb12 (SEQ ID NOs: 343 + 284) Bispecific2 = bNAb12_HC_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 344 + 284) Bispecific3 = D1m-K8C-G99C_1xG4S_LC-bNAb12 (SEQ ID NOs: 283 + 345) Bispecific4 = bNAb12_LC_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 283 + 346) Combo = bNAb12 (SEQ ID NOs: 283 + 284) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb12 (SEQ ID NOs: 283 + 284) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(283) TABLE-US-00019 TABLE 15 IC50 (nM) of different bNAb13-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb13-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 42368 0.24 1.52 >500 1.58 X2088.C9 2.05 5.18 >500 9.20 ZM106.9 0.28 3.04 22.43 15.14 3326_V4_C3 52.07 191.00 >500 >500 3637_V5_C3 37.30 266.00 >500 >500 3468_V1_C12 4.25 25.00 >500 64.15 Q461_E2 0.50 4.14 >500 11.76 CC1/85 0.19 * * 1.98 620345_C1 3.58 * >500 99.28 0260.v5.c36 0.36 * 18.10 119.29 TH976_17 37.26 * >500 39.05 928_28 0.79 * 201.39 0.77 YU-2 0.17 * 48.30 0.20 6471_V1_C16 0.16 * 133.35 5.07 93UG065 0.58 * 46.41 142.61 NL4-3 0.21 * * 0.31 T278.50 0.34 * * 1.43 CAP45.2.00.G3 2.26 * * 20.48 HIV-2-HCC-01 11.13 * * >300 * not tested Table 15 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb13 (SEQ ID NOs: 347 + 289) Combo = bNAb13 (SEQ ID NOs: 288 + 289) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb13 (SEQ ID NOs: 288 + 289) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(284) TABLE-US-00020 TABLE 16 IC50 (nM) of different bNAb14-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb14-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 X2088.C9 1.26 1.73 2.79 9.20 ZM106.9 0.62 1.08 1.26 15.14 3637_V5_C3 22.85 84.20 >500 >500 3468_V1_C12 17.35 35.05 >500 64.15 Q461_E2 2.77 11.40 >500 11.76 3326_V4_C3 196.50 >500 >500 >500 42368 1.15 1.84 >500 1.58 CC1/85 1.74 * * 1.98 NL4-3 0.29 * * 0.31 T278.50 6.81 * * 1.43 CAP45.2.00.G3 2.85 * * 20.48 HIV-2-HCC-01 98.75 * * >300 * not tested Table 16 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb14 (SEQ ID NOs: 348 + 294) Combo = bNAb14 (SEQ ID NOs: 293 + 294) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb14 (SEQ ID NOs: 293 + 294) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(285) TABLE-US-00021 TABLE 17 IC50 (nM) of different bNAb15-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb15-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 X2088.C9 1.19 0.95 2.58 9.20 ZM106.9 0.68 0.40 1.34 15.14 3637_V5_C3 25.45 5.33 5.00 >500 3468_V1_C12 22.00 35.80 >500 64.15 Q461_E2 2.81 5.98 >500 11.76 3326_V4_C3 142.00 >500 >500 >500 42368 1.11 1.55 >500 1.58 CC1/85 1.65 * * 1.98 NL4-3 0.36 * * 0.31 T278.50 7.66 * * 1.43 CAP45.2.00.G3 4.48 * * 20.48 HIV-2-HCC-01 142.16 * * >300 * not tested Table 17 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb15 (SEQ ID NOs: 349 + 299) Combo = bNAb15 (SEQ ID NOs: 298 + 299) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb15 (SEQ ID NOs: 298 + 299) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(286) TABLE-US-00022 TABLE 18 IC50 (nM) of different bNAb16-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb16-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 X2088.C9 0.25 2.02 289.00 9.20 ZM106.9 2.42 11.63 >500 15.14 3637_V5_C3 19.15 129.50 >500 >500 3468_V1_C12 11.21 26.75 >500 64.15 Q461_E2 2.03 6.06 >500 11.76 3326_V4_C3 31.75 148.50 >500 >500 42368 0.69 1.92 >500 1.58 CC1/85 0.56 * * 1.98 NL4-3 0.14 * * 0.31 T278.50 2.29 * * 1.43 CAP45.2.00.G3 2.08 * * 20.48 HIV-2-HCC-01 237.46 * * >300 * not tested Table 18 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb16 (SEQ ID NOs: 350 +304) Combo = bNAb16 (SEQ ID NOs: 303 + 304) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb16 (SEQ ID NOs: 303 + 304) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(287) TABLE-US-00023 TABLE 19 IC50 (nM) of different bNAb17-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb17-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 X2088.C9 0.84 2.44 >500 9.20 ZM106.9 8.48 9.13 >500 15.14 3637_V5_C3 45.75 97.90 >500 >500 3468_V1_C12 39.60 27.85 >500 64.15 Q461_E2 6.70 6.03 >500 11.76 3326_V4_C3 287.50 171.50 >500 >500 42368 2.14 1.50 >500 1.58 CC1/85 0.91 * * 1.98 NL4-3 0.43 * * 0.31 T278.50 4.25 * * 1.43 CAP45.2.00.G3 1.39 * * 20.48 HIV-2-HCC-01 300.00 * * >300 * not tested Table 19 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb17 (SEQ ID NOs: 351 + 309) Combo = bNAb17 (SEQ ID NOs: 308 + 309) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb17 (SEQ ID NOs: 308 + 309) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(288) TABLE-US-00024 TABLE 20 IC50 (nM) of different bNAb18-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb18-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 X2088.C9 0.63 0.26 0.20 9.20 ZM106.9 0.25 0.08 0.11 15.14 3637_V5_C3 102.85 377.00 >500 >500 3468_V1_C12 26.60 38.60 >500 64.15 Q461_E2 3.16 9.98 >500 11.76 3326_V4_C3 157.00 369.50 >500 >500 42368 0.94 2.18 >500 1.58 CC1/85 2.06 * * 1.98 NL4-3 0.33 * * 0.31 T278.50 5.83 * * 1.43 CAP45.2.00.G3 20.89 * * 20.48 HIV-2-HCC-01 93.74 * * >300 * not tested Table 20 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb18 (SEQ ID NOs: 352 + 314) Combo = bNAb18 (SEQ ID NOs: 313 + 314) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb18 (SEQ ID NOs: 313 + 314) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(289) TABLE-US-00025 TABLE 21 IC50 (nM) of different bNAb19-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) bNAb19-Derived Molecules CD4 Controls Envelope Bispecific Combo mAb CD4 X2088.C9 1.63 4.40 >300 9.20 ZM106.9 3.91 8.27 >300 15.14 3637_V5_C3 16.50 168.00 >300 >500 3468_V1_C12 4.56 24.54 >300 64.15 Q461_E2 3.18 4.49 >300 11.76 3326_V4_C3 130.00 241.67 >300 >500 42368 1.16 1.10 >300 1.58 CC1/85 0.52 * * 1.98 NL4-3 0.23 * * 0.31 T278.50 0.79 * * 1.43 CAP45.2.00.G3 5.54 * * 20.48 HIV-2-HCC-01 6.54 * * >300 * not tested Table 21 Molecule Key: Bispecific = D1m-K8C-G99C_1xG4S_bNAb19 (SEQ ID NOs: 353 + 319) Combo = bNAb19 (SEQ ID NOs: 318 + 319) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) mAb = bNAb19 (SEQ ID NOs: 318 + 319) CD4 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(290) TABLE-US-00026 TABLE 22 IC50 (nM) of different bNAb20/21/22-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb20-Derived Molecules bNAb21-Derived Molecules bNAb22-Derived Molecules Controls Envelope 1 2 3 4 5 6 7 8 9 10 X2088.C9 0.03 0.04 0.04 0.02 0.02 0.02 0.04 0.03 0.04 9.20 ZM106.9 0.01 0.02 0.02 0.01 0.01 0.01 0.01 0.01 0.01 15.14 3637_V5_C3 6.2 21.9 61.6 4.7 117.0 326.0 6.8 115.0 484.0 >500 3468_V1_C12 0.01 0.02 0.02 0.01 0.01 0.01 0.01 0.02 0.01 64.15 Q461_E2 0.9 5.9 72.1 0.4 10.0 186.0 2.0 7.1 >500 11.76 3326_V4_C3 1.3 24.1 24.0 0.3 14.5 12.5 0.6 6.1 16.0 >500 42368 0.5 3.7 59.4 0.2 1.9 128.0 0.7 1.2 391.0 1.58 Table 22 Molecule Key: 1 = D1m-K8C-G99C_1xG4S_bNAb20 (SEQ ID NOs: 354 + 324) 2 = bNAb20 (SEQ ID NOs: 323 + 324) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 3 = bNAb20 (SEQ ID NOs: 323 + 324) 4 = D1m-K8C-G99C_1xG4S_bNAb21 (SEQ ID NOs: 355 + 329) 5 = bNAb21 (SEQ ID NOs: 328 + 329) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 6 = bNAb21 (SEQ ID NOs: 328 + 329) 7 = D1m-K8C-G99C_1xG4S_bNAb22 (SEQ ID NOs: 356 + 366) 8 = bNAb22 (SEQ ID NOs: 332 + 366) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 9 = bNAb22 (SEQ ID NOs: 332 + 366) 10 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
(291) TABLE-US-00027 TABLE 23 IC50 (nM) of different bNAb23-derived bispecific formats and control molecules against a panel of HIV-1 envelopes in PSV assay (ACTOne cells) CD4 bNAb23-Derived Molecules Controls Envelope 1 2 3 4 5 6 X2088.C9 6.05 0.09 12.05 5.89 >500 9.20 ZM106.9 3.96 0.12 6.06 15.06 17.4 15.14 3637_V5_C3 129.50 2.26 >500 294.00 >500 >500 3468_V1_C12 62.10 6.17 384.00 53.05 >500 64.15 Q461_E2 4.29 0.80 116.00 7.11 >500 11.76 3326_V4_C3 44.90 3.94 108.00 173.00 15.3 >500 42368 0.62 0.30 10.70 1.57 >500 1.58 Table 23 Molecule Key: 1 = D1m-K8C-G99C_1xG4S_bNAb23 (SEQ ID NOs: 357 + 337) 2 = D1m-K8C-G99C_1xG4S_bNAb23-LC (SEQ ID NOs: 336 + 358) 3 = bNAb23-HC_1xG4S_D1m-K8C-G99C (SEQ ID NOs: 359 + 337) 4 = bNAb23 (SEQ ID NOs: 336 + 337) + D1m-K8C-G99C_Fc (SEQ ID NO: 158) (combo) 5 = bNAb23 (SEQ ID NOs: 336 + 337) 6 = D1m-K8C-G99C_Fc (SEQ ID NO: 158)
Example 8Broad Spectrum Anti-Viral Activity of a bNAb1-derived Bispecific Molecule (SEQ ID NO:121 and SEO ID NO: 63)
(292) A selected bispecific molecule derived from bNAb1 having two heavy chains (SEQ ID NO:121) and two light chains (SEQ ID NO:63) was independently tested against an external panel of pseudo-typed HIV-1 virus containing 119 HIV-1 envelopes and 1 control envelope in a PSV assay 15 (TMZ.blsee Example 3 above), to further evaluate its breadth and potency. As shown in Table 24, the bispecific molecule completely and potently inhibited all of the 119 HIV-1 envelopes in this assay.
(293) TABLE-US-00028 TABLE 24 IC50 and IC80 values (g/ml) of bNAb1-derived bispecific molecule (SEQ ID NO: 121 and SEQ ID NO: 63) against 119 HIV-1 envelopes in PSV assay Virus ID Clade* IC50 IC80 MPI 6535.3 B 0.007 0.021 100 QH0692.42 B 0.020 0.066 100 SC422661.8 B 0.013 0.032 100 PVO.4 B 0.010 0.037 100 TRO.11 B 0.011 0.026 100 AC10.0.29 B 0.006 0.022 100 RHPA4259.7 B 0.007 0.021 100 THRO4156.18 B 0.027 0.095 100 REJO4541.67 B 0.022 0.054 100 TRJO4551.58 B 0.006 0.022 100 WITO4160.33 B 0.024 0.099 100 CAAN5342.A2 B 0.023 0.085 100 WEAU_d15_410_787 B (T/F) 0.009 0.033 100 1006_11_C3_1601 B (T/F) 0.005 0.014 100 1054_07_TC4_1499 B (T/F) 0.014 0.041 100 1056_10_TA11_1826 B (T/F) 0.009 0.026 100 1012_11_TC21_3257 B (T/F) 0.004 0.015 100 6240_08_TA5_4622 B (T/F) 0.019 0.054 100 6244_13_B5_4576 B (T/F) 0.011 0.039 100 62357_14_D3_4589 B (T/F) 0.021 0.060 100 SC05_8C11_2344 B (T/F) 0.016 0.045 100 Du156.12 C 0.005 0.018 100 Du172.17 C 0.010 0.029 100 Du422.1 C 0.011 0.044 100 ZM197M.PB7 C 1.148 5.807 100 ZM214M.PL15 C 0.020 0.069 100 ZM233M.PB6 C 0.046 0.171 100 ZM249M.PL1 C 0.006 0.021 100 ZM53M.PB12 C 0.275 0.908 100 ZM109F.PB4 C 0.036 0.132 100 ZM135M.PL10a C 0.030 0.119 100 CAP45.2.00.G3 C 0.260 2.976 100 CAP210.2.00.E8 C 0.023 0.102 100 HIV-001428-2.42 C 0.002 0.004 100 HIV-0013095-2.11 C 0.016 0.042 100 HIV-16055-2.3 C 0.044 0.159 100 HIV-16845-2.22 C 0.042 0.147 100 Ce1086_B2 C (T/F) 0.085 0.297 100 Ce0393_C3 C (T/F) 0.008 0.022 100 Ce1176_A3 C (T/F) 0.010 0.028 100 Ce2010_F5 C (T/F) 0.206 0.693 100 Ce0682_E4 C (T/F) 0.080 1.232 100 Ce1172_H1 C (T/F) 0.005 0.015 100 Ce2060_G9 C (T/F) 0.016 0.059 100 Ce703010054_2A2 C (T/F) 0.342 1.030 100 BF1266.431a C (T/F) 0.106 1.283 100 246F C1G C (T/F) 0.009 0.031 100 249M B10 C (T/F) 0.627 5.222 99 ZM247v1(Rev-) C (T/F) 0.004 0.014 100 7030102001E5(Rev-) C (T/F) 0.010 0.025 100 1394C9G1(Rev-) C (T/F) 0.008 0.028 100 Ce704809221_1B3 C (T/F) 0.014 0.060 100 CNE19 BC 0.054 0.270 100 CNE20 BC 0.001 0.004 100 CNE21 BC 0.004 0.013 100 CNE17 BC 0.023 0.073 100 CNE30 BC 0.026 0.074 100 CNE52 BC 0.018 0.043 100 CNE53 BC 0.005 0.013 100 CNE58 BC 0.008 0.022 100 MS208.A1 A 1.457 6.230 100 Q23.17 A 0.010 0.025 100 Q461.e2 A 0.025 0.086 100 Q769.d22 A 0.046 0.122 100 Q259.d2.17 A 0.167 0.459 100 Q842.d12 A 0.006 0.015 100 0260.v5.c36 A 0.031 0.111 100 3415.v1.c1 A 0.010 0.028 100 3365.v2.c20 A 0.011 0.036 100 191955_A11 A (T/F) 0.032 0.115 100 191084 B7-19 A (T/F) 0.009 0.027 100 9004SS_A3_4 A (T/F) 0.008 0.023 100 T257-31 CRF02_AG 0.022 0.064 100 928-28 CRF02_AG 0.029 0.086 100 263-8 CRF02_AG 0.011 0.043 100 T250-4 CRF02_AG 0.005 0.012 100 T251-18 CRF02_AG 0.023 0.064 100 T278-50 CRF02_AG 0.010 0.027 100 T255-34 CRF02_AG 0.025 0.091 100 211-9 CRF02_AG 0.025 0.085 100 235-47 CRF02_AG 0.170 0.800 100 620345.c01 CRF01_AE 0.247 2.737 100 CNE8 CRF01_AE 0.007 0.021 100 C1080.c03 CRF01_AE 0.014 0.027 100 R2184.c04 CRF01_AE 0.006 0.022 100 R1166.c01 CRF01_AE 0.252 1.001 100 R3265.c06 CRF01_AE 0.065 0.254 100 C2101.c01 CRF01_AE 0.005 0.016 100 C3347.c11 CRF01_AE 0.006 0.015 100 C4118.c09 CRF01_AE 0.512 4.356 100 CNE5 CRF01_AE 0.014 0.038 100 BJOX009000.02.4 CRF01_AE 0.006 0.020 100 BJOX015000.11.5 CRF01_AE (T/F) 0.013 0.035 100 BJOX010000.06.2 CRF01_AE (T/F) 0.035 0.114 100 BJOX025000.01.1 CRF01_AE (T/F) 0.035 0.140 100 BJOX028000.10.3 CRF01_AE (T/F) 0.004 0.010 100 X1193_c1 G 0.015 0.034 100 P0402_c2_11 G 0.005 0.019 100 X1254_c3 G 0.014 0.040 100 X2088_c9 G 0.017 0.048 100 X2131_C1_B5 G 0.014 0.039 100 P1981_C5_3 G 0.014 0.038 100 X1632_S2_B10 G 0.027 0.097 100 3016.v5.c45 D 0.041 0.117 100 A07412M1.vrc12 D 0.009 0.034 100 231965.c01 D 0.023 0.068 100 231966.c02 D 0.006 0.018 100 6405.v4.c34 D 0.029 0.082 100 3817.v2.c59 CD 0.025 0.063 100 6480.v4.c25 CD 0.008 0.024 100 6952.v1.c20 CD 0.037 0.109 100 6811.v7.c18 CD 0.004 0.014 100 89-F1_2_25 CD 0.035 0.110 100 3301.v1.c24 AC 0.005 0.018 100 6041.v3.c23 AC 0.109 0.553 100 6540.v4.c1 AC 0.002 0.012 100 6545.v4.c1 AC 0.011 0.031 100 0815.v3.c3 ACD 0.003 0.008 100 3103.v3.c10 ACD 0.012 0.027 100 MuLV Negative >50 >50 0 Control
Example 9Activity of bNAb1-Derived Bispecific Molecules Against Laboratory and Clinical Isolates of HIV-1 in Replicating Virus Assays
(294) Method
(295) Clinical and Laboratory Isolates
(296) All clinical and laboratory isolates were originally obtained from the NIH AIDS Reagent Program (currently NIH HIV Reagent Program, https:hivreagentprogram.org/). The proviral clone of NL.sub.4-3 (obtained from NIH) was used to make the replicating reporter virus NLRepRluc, in which a section of the nef gene from the proviral clone of NL.sub.4-3 was replaced with the Renilla luciferase gene. Virus was produced through transfection of HEK293T cells using Lipofectamine Plus (Invitrogen, Carlsbad, CA), according to the manufacturer's instructions. The replication-competent virus was harvested 3 days after transfection of HEK 293T cells with the modified pNLRepRluc proviral clone and titrated in MT-2 cells using luciferase activity as a biomarker.
(297) Clinical isolates were initially propagated in human PBMC cells. T-tropic laboratory virus strains IIIB, NL.sub.4-3, HXB2, LAI, MN and RF viruses were propagated in MT-2 cells, while M-tropic laboratory strains Bal and JR-FL were propagated in PM1 cells. Titers of virus stocks were determined in PBMC using a virus infectivity assay with a p24 antigen endpoint (p24 ELISA kit; PerkinElmer Life Sciences). All those viruses were further titered in MT2 or CCR5-B6 cells before experiments by using luciferase enzyme activity as an endpoint for 50% tissue culture infectious dose (TCID50) determination.
(298) Cells
(299) MT-2 cells were obtained from the American Type Culture Collection (ATCC) and were propagated in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 units/ml of penicillin G, 100 g/ml of streptomycin, 10 mM HEPES buffer pH 7.55 and 2-mM L-glutamine. HEK293T cells were ere obtained from the ATCC and propagated in DMEM media supplemented with 10% heat-inactivated FBS. The ACTOne cells were originally derived from HEK293T cells and express CD4, CCR5 and CXCR4. They are grown in DMEM media supplemented with 10% heat-inactivated FBS, 100 U/ml of penicillin G, 100 g/ml of streptomycin, 5 g/ml blasticidin, 200 g/ml G418 and 1.5 g/ml puromycin. CCR5-B6 cells were generated in-house at ViiV Branford CT USA. To generate CCR5-B6 cells, human CCR5 lentiviral particles were used to infect the MT4-B6 cells (obtained from Bristol-Myers Squibb) that has an integrated copy of the LTR-fire-fly luciferase reporter (backbone: plenti-P2A-Puro, RC223291L3V, Origene) and stable cells were selected by using G418 (0.6 mg/ml) and puromycin (2 g/ml). The CCR5-B6 cells express firefly luciferase from an HIV-1 LTR promoter after infection with HIV. They are grown in RPMI 1640 supplemented with 10% heat inactivated fetal bovine serum (FBS), 10 mM HEPES buffer pH 7.55, 2 mM L-glutamine, 100 units/ml penicillin G, and 100 g/ml streptomycin, 2 g/ml puromycin, 0.6 mg/ml G418.
(300) Replicating Virus Assay
(301) Assay Using NLRepRluc Virus
(302) The NLRepRluc was used to infect MT-2 cells at a multiplicity of 0.01 for 1 hour before adding the proteins to the 96-well plates. Antibodies were serially diluted four-fold and 11 concentrations were plated in triplicate. After 4 days of incubation, cells were processed and quantitated for virus growth by the amount of expressed luciferase. Luciferase was quantitated using the ENDUREN substrate from Promega (Madison, WI) according to the manufacturer's instructions. Luciferase activity was measured immediately on an ENVISION multilabel plate reader (PerkinElmer, Waltham MA). EC.sub.50 values were calculated by comparing the amount of luciferase produced in the presence of antigen binding protein compared to wells where no antigen binding protein (DMSO control) was added. A 5-parameter sigmoidal equation was used to fit the resulting signal vs. concentration curves, and the concentration of each antigen binding protein that produced 50% maximal inhibition (EC.sub.50) was determined. The results of three independent experiments were averaged and plotted, with error bars corresponding to 1 standard deviation.
(303) Replicating Virus Assay Using Laboratory Strains and Clinical Isolates
(304) Replicating laboratory strains and clinical isolates were prepared as described above. MT2 cells or CCR5-B6 cells were resuspended in corresponding media and distributed to 96-well assay plates (26,000 cells/well in 100 uL; Corning, Tewksbury, MA) containing serial dilutions of inhibitors in DMSO (5 or 3-fold dilutions, columns 1-10). The blank controls were wells containing DMSO (column 11, 12). Replicating whole viruses of laboratory strains or clinic isolates were diluted in RPMI-1640 culture medium based on 50% tissue culture infectious dose (TCID50) determination such that undiluted stock virus was added to the first well, and 100 l of viral culture medium were loaded to the wells already containing compounds and cells (total 200 l/well), resulting in a final concentration of DMSO of 1%. Plates were incubated at 37 C. and 5% CO.sub.2 for approximately 4 days. After that, Renilla luciferase activity was measured (Enduren reagent, Promega Corp., Madison, WI) on an EnVision Multilabel pate reader (Perkin Elmer, Inc., Waltham MA). The 50% effective concentration (EC.sub.50) was calculated by using the exponential form of the median effect equation where (Fa)=1/[1+ (ED.sub.50/drug conc.)].
(305) Results
(306) Table 25.1 shows that the bNAb1-derived bispecific molecules are consistently about 10-fold more active than the mixture of the component parts (bNAb1 and CD4 domain), again indicating a clear anti-viral synergy result from fusing these component binding domains.
(307) TABLE-US-00029 TABLE 25.1 EC50 (nM) values of bNAb1-derived bispecific and control molecules against NL4-3 in a replicating virus assay Molecules (SEQ ID NO) EC50 (nM) D1m_1xG4S_bNAb1 (102 + 63) 0.03 D1m_2xG4S_bNAb1 (103 + 63) 0.03 D1m_3xG4S_bNAb1 (104 + 63) 0.02 D1m_4xG4S_bNAb1 (105 + 63) 0.03 D1m_His (4*) + bNAb1 (62 + 63) 0.22 D1m_His (4*) 0.23 bNAb1 (62 + 63) >500 *plus a 6xHis tag (six C-terminal histidine residues)
(308) A bNAb1-derived bispecific molecule (D1m-K8C-G99C_1xG4S_bNAb1, SEQ ID NOS: 121+63) was tested against a panel of 13 clinical and 8 laboratory HIV-1 isolates in a replicating virus assay using MT-2 and CCR5-B6 cells. As shown in Table 25.2, this molecule neutralized all strains, with EC50s less than 1 nM (geometric mean EC50 being 0.11 nM against clinical isolates and 0.26 nM against lab strains), demonstrating again its strong potency and breadth of activity.
(309) TABLE-US-00030 TABLE 25.2 EC50 (nM) of a bNAb1-derived bispecific against a panel of HIV-1 clinical isolates and laboratory strains in a replicating virus assay bNAb1-Derived Isolates Clade Bispecific Clinical isolates 93US141 B 0.04 93US144 B 0.07 ASM34 B 0.15 BK132 B 0.19 BZ167 B 0.05 CC1/85 B 0.2 CM237 B 0.03 ETH2220 C 0.08 I-2496 A 0.15 SE364 C 0.2 UG268 C 0.06 UG270 D 0.42 US4 B 0.39 Laboratory strains HXB2 0.04 BaL 0.83 IIIB/H9 0.56 IIIB/HOS 0.56 JR-FL 0.04 LAI 0.39 MIN 0.15 NL4-3 0.39 RF 0.56 Table 25.2 Molecule Key: 1 = D1m-K8C-G99C_1xG4S_bNAb1 (SEQ ID NOs: 121 + 63)
Example 10In Vitro Resistance Barrier
(310) To assess the resistance barrier of the bispecific molecules, we examined the relative rates at which HIV NL4-3 virus can escape inhibition by a panel of antibodies versus a DMSO control.
(311) MT2 cells (2.010.sup.5/well in RPMI 1640+50 mg/ml penicillin and streptomycin+10 mM HEPES buffer pH 7.55+2 mM L-glutamine. +0.2% DMSO) were pre-infected at an MOI (multiplicity of infection) of 0.005 for 2.5 hours and then pelleted to remove unbound virus particles. One mL of infected cells was added to each well of a 24 well plate. Replicates of antibody dilutions at the concentrations of 20-, 30-, or 40-fold of IC50 values were then added (1 mL of a 2 stock) to achieve 2 mL assay volume.
(312) Every 3-4 days images of the wells were captured and 1 ml of each well is removed and replaced by fresh preparations of each condition. This process was continued until there was viral breakthrough (observed cytopathic effect (CPE)>80%) or until there was a confirmed elimination of infected cells (via challenged elimination). When either condition was achieved, the sample was collected as pellet and supernatant (via centrifugation) and stored at 80 C. until genotypic analysis was performed to confirm the presence of resistance mutations. The days from infection to breakthrough were used to estimate the resistance barrier of a certain molecule. The longer it takes the virus to develop CPE, which indicates resistance, the higher the resistance barrier is.
(313) Table 26 shows that, in this experimental setting, the bNAb1-derived bispecific molecule (SEQ ID NO: 105 and SEQ ID NO:63) exhibited a much higher resistance barrier than the soluble CD4 domain or bNAb1 alone at all concentrations. This again indicates synergy between soluble CD4 domains and bNAb1 when fused together.
(314) TABLE-US-00031 TABLE 26 Average days Group Molecule Conc. (nM) before CPE 20x EC50 D1m_4xG4S_bNAb1 4.2 16 D1m_His 8.4 4 bNAb1 18 4 30x EC50 D1m_4xG4S_bNAb1 6.3 54 D1m_His 12.5 5 bNAb1 27 4 40x EC50 D1m_4xG4S_bNAb1 8.4 62 D1m_His 16.7 7 bNAb1 36 4 Table 26 Molecule Key: D1m_4xG4S_bNAb1 = SEQ ID NO: 105 and 63 D1m_His = SEQ ID NO: 4 plus a 6xHis tag (six C-terminal histidine residues) bNAb1 = SEQ ID NOs: 62 and 63
Example 11Anti-HIV-2 Activity of a bNAb1-Derived Bispecific Molecule (SEQ ID NO: 121 and SEQ ID NO:63) in Pseudotyped and Replicating Virus Assays
(315) HIV-2 differs from HIV-1 in that it originates from the transmission of simian immunodeficiency virus (SIV) from sooty mangabeys (SIVsmm) to human (Gao et al, J Virol. 1994; 68 (11): 7433-7447) whilst HIV-1 stems from the transmission from chimpanzees and western gorillas (HIV-1 group M and O, respectively). HIV-2 can also cause AIDS but is far less pathogenic and wide-spread than HIV-1 (de Silva et al, Trends Microbiol. 2008; 16 (12): 588-595; Da Silva et al, AIDS. 2008; 22 (10): 1195-1202). Though also using CD4 for infection (Sattentau et al, AIDS. 1988; 2 (2): 101-105), HIV-2 shares only 40% identity in the gp160 amino acid sequence with HIV-1, and is therefore less sensitive or insensitive to HIV-1 envelope-directed bnAbs (Kong et al, JVI 2012; 86 (2): 947-960). The bNAb1-derived bispecific molecule (SEQ ID NO:121 and SEQ ID NO: 63) showed strong potency and anti-viral synergy against the 2 HIV-2 Env-pseudotypes tested in PSV assays and the laboratory NIHZ strain examined in a replicating virus assay (Table 27), illustrating its exceptional breadth of anti-HIV activity and excellent synergy.
(316) TABLE-US-00032 TABLE 27 IC50 (nM) of a bNAb1-derived bispecific and control molecules against HIV-2 strains bNAb1-derived molecules Strain Assay 1 2 3 4 HIV-2-ATM88 PSV 0.07 3.93 >500 10.53 HIV-2-HCC-01 PSV 0.78 >500 >500 >500 HIV-2 NIHZ Replicating 2.68 * * * virus * not tested Table 27 Molecule Key: 1 = D1m-K8C-G99C_1xG4S_bNAb1 (SEQ ID NOs: 121 + 63) 2 = D1m_His (SEQ ID NO: 4*) 3 = bNAb1 (SEQ ID NOs: 62 + 63) 4 = D1m_His (SEQ ID NO: 4*) + bNAb1(SEQ ID NOs: 62 + 63) (combo) *plus a 6xHis tag (six C-terminal histidine residues)
(317) TABLE-US-00033 SEQUENCE LISTING SEQ ID NO Name/Identifier Description 1 D1D2 Human CD4 D1D2 (wild type) 2 D1mD2 Human CD4 mD1.22-D2 (D1 mutations S23N, A55V, and L96V) 3 D1 Human CD4 D1 (wild type) 4 D1m Human CD4 mD1.22 (NCBI accession code: QHY83614.1; D1 mutations L5Y, S23N, A55V, I76P, L96V and F98V) 5 D1m-K8V Human CD4 mD1.22 + K8V 6 D1m-E91Q Human CD4 mD1.22 + E91Q 7 D1m-E91H Human CD4 mD1.22 + E91H 8 D1m-E87G Human CD4 mD1.22 + E87G 9 D1m-N52W Human CD4 mD1.22 + N52W 10 D1m-K8I Human CD4 mD1.22 + K8I 11 D1m-K8C-G99C Human CD4 mD1.22 + K8C, G99C 12 D1m-T11C-K72C Human CD4 mD1.22 + T11C, K72C 13 D1m-E13C-I70C Human CD4 mD1.22 + E13C, I70C 14 D1m-H27C-G38C Human CD4 mD1.22 + H27C, G38C 15 D1m-K21C-G65C Human CD4 mD1.22 + K21C, G65C 16 D1m-Q25E Human CD4 mD1.22 + Q25E 17 D1m-H27D Human CD4 mD1.22 + H27D 18 D1m-R58V Human CD4 mD1.22 + R58V 19 D1m-R58N Human CD4 mD1.22 + R58N 20 D1m-R58T Human CD4 mD1.22 + R58T 21 D1m-L61M Human CD4 mD1.22 + L61M 22 bNAb1 CDRH1 CDRH1 of bNAb1 23 bNAb1 CDRH2 CDRH2 of bNAb1 24 bNAb1 CDRH3 CDRH3 of bNAb1 25 bNAb1 CDRL1 CDRL1 of bNAb1 26 bNAb1 CDRL2 CDRL2 of bNAb1 27 bNAb1 CDRL3 CDRL3 of bNAb1 28 bNAb2 CDRH1 CDRH1 of bNAb2 29 bNAb2 CDRH2 CDRH2 of bNAb2 30 bNAb2 CDRH3 CDRH3 of bNAb2 31 bNAb2 CDRL1 CDRL1 of bNAb2 32 bNAb2 CDRL2 CDRL2 of bNAb2 33 bNAb2 CDRL3 CDRL3 of bNAb2 34 bNAb3 CDRH1 CDRH1 of bNAb3 35 bNAb3 CDRH2 CDRH2 of bNAb3 36 bNAb3 CDRH3 CDRH3 of bNAb3 37 bNAb3 CDRL1 CDRL1 of bNAb3 38 bNAb3 CDRL2 CDRL2 of bNAb3 39 bNAb3 CDRL3 CDRL3 of bNAb3 40 bNAb4 CDRH1 CDRH1 of bNAb4 41 bNAb4 CDRH2 CDRH2 of bNAb4 42 bNAb4 CDRH3 CDRH3 of bNAb4 43 bNAb4 CDRL1 CDRL1 of bNAb4 44 bNAb4 CDRL2 CDRL2 of bNAb4 45 bNAb4 CDRL3 CDRL3 of bNAb4 46 bNAb5 CDRH1 CDRH1 of bNAb5 47 bNAb5 CDRH2 CDRH2 of bNAb5 48 bNAb5 CDRH3 CDRH3 of bNAb5 49 bNAb5 CDRL1 CDRL1 of bNAb5 50 bNAb5 CDRL2 CDRL2 of bNAb5 51 bNAb5 CDRL3 CDRL3 of bNAb5 52 bNAb6 CDRH1 CDRH1 of bNAb6 53 bNAb6 CDRH2 CDRH2 of bNAb6 54 bNAb6 CDRH3 CDRH3 of bNAb6 55 bNAb6 CDRL1 CDRL1 of bNAb6 56 bNAb6 CDRL2 CDRL2 of bNAb6 57 bNAb6 CDRL3 CDRL3 of bNAb6 58 bNAb1 VH Heavy chain variable region of bNAb1 59 bNAb1 VL Light chain variable region of bNAb1 60 bNAb1* VL Light chain variable region of bNAb1 with F32Y mutation 61 bnAb1 HC Full heavy chain of bNAb1 62 bNAb1 HC + LS Full heavy chain of bNAb1 with M428L/N434S mutations 63 bNAb1 LC Full light chain of bNAb1 64 bNAb1* LC Full light chain of bNAb1 with F32Y mutation 65 bNAb2 VH Heavy chain variable region of bNAb2 66 bNAb2 VL Light chain variable region of bNAb2 67 bNAb2 HC Full heavy chain of bNAb2 68 bNAb2 HC + LS Full heavy chain of bNAb2 with M428L/N434S mutations 69 bNAb2 LC Full light chain of bNAb2 70 bNAb3 VH Heavy chain variable region of bNAb3 71 bNAb3 VL Light chain variable region of bNAb3 72 bNAb3 HC Full heavy chain of bNAb3 73 bNAb3 HC + LS Full heavy chain of bNAb3 with M428L/N434S mutations 74 bNAb3 LC Full light chain of bNAb3 75 bNAb4 VH Heavy chain variable region of bNAb4 76 bNAb4 VL Light chain variable region of bNAb4 77 bNAb4 HC Full heavy chain of bNAb4 78 bNAb4 HC + LS Full heavy chain of bNAb4 with M428L/N434S mutations 79 bNAb4 LC Full light chain of bNAb4 80 bNAb5 VH Heavy chain variable region of bNAb5 81 bNAb5 VL Light chain variable region of bNAb5 82 bNAb5 HC Full heavy chain of bNAb5 83 bNAb5 HC + LS Full heavy chain of bNAb5 with M428L/N434S mutations 84 bNAb5 LC Full light chain of bNAb5 85 bNAb6 VH Heavy chain variable region of bNAb6 86 bNAb6 VL Light chain variable region of bNAb6 87 bNAb6 HC Full heavy chain of bNAb6 88 bNAb6 HC + LS Full heavy chain of bNAb6 with M428L/N434S mutations 89 bNAb6 LC Full light chain of bNAb6 90 1xG4S Linker 91 2xG4S Linker 92 3xG4S Linker 93 4xG4S Linker 94 5xG4S Linker 95 6xG4S Linker 96 D1mD2_0xG4S_bNAb1-HC Human CD4 mD1.22-D2 fused to the N- terminus of bNAb1 heavy chain with no linker, Fc includes M428L/N434S 97 D1mD2_1xG4S_bNAb1-HC Human CD4 mD1.22-D2 fused to the N- terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 98 D1mD2_2xG4S_bNAb1-HC Human CD4 mD1.22-D2 fused to the N- terminus of bNAb1 heavy chain with 2xG4S linker, Fc includes M428L/N434S 99 D1mD2_3xG4S_bNAb1-HC Human CD4 mD1.22-D2 fused to the N- terminus of bNAb1 heavy chain with 3xG4S linker, Fc includes M428L/N434S 100 D1mD2_4xG4S_bNAb1-HC Human CD4 mD1.22-D2 fused to the N- terminus of bNAb1 heavy chain with 4xG4S linker, Fc includes M428L/N434S 101 D1m_0xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with no linker, Fc includes M428L/N434S 102 D1m_1xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 103 D1m_2xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with 2xG4S linker, Fc includes M428L/N434S 104 D1m_3xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with 3xG4S linker, Fc includes M428L/N434S 105 D1m_4xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with 4xG4S linker, Fc includes M428L/N434S 106 D1m_5xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with 5xG4S linker, Fc includes M428L/N434S 107 D1m_6xG4S_bNAb1-HC Human CD4 mD1.22 fused to the N- terminus of bNAb1 heavy chain with 6xG4S linker, Fc includes M428L/N434S 108 D1m_0xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with no linker 109 D1m_1xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with 1xG4S linker 110 D1m_2xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with 2xG4S linker 111 D1m_3xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with 3xG4S linker 112 D1m_4xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with 4xG4S linker 113 D1m_5xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with 5xG4S linker 114 D1m_6xG4S_bNAb1-LC Human CD4 mD1.22 fused to the N- terminus of bNAb1 light chain with 6xG4S linker 115 D1m-K8C-G99C_1xG4S_bNAb1-LC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb1 light chain with 1xG4S linker 116 bNAb1-HC-mid_1xG4S_D1m-K8C- Human CD4 mD1.22 + K8C + G99C fused in G99C between the CH1 domain and hinge of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 117 bNAb1-HC_1xG4S_D1m-K8C-G99C Human CD4 mD1.22 + K8C + G99C fused to the C-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 118 bNAb1-LC_1xG4S_D1m-K8C-G99C Human CD4 mD1.22 + K8C + G99C fused to the C-terminus of bNAb1 light chain with 1xG4S linker 119 D1m-K8I_1xG4S_bNAb1-HC Human CD4 mD1.22 + K8I fused to the N- terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 120 D1m-K8V_1xG4S_bNAb1-HC Human CD4 mD1.22 + K8V fused to the N- terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 121 D1m-K8C-G99C_1xG4S_bNAb1-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 122 D1m-T11C-K72C_1xG4S_bNAb1- Human CD4 mD1.22 + T11C + K72C fused HC to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 123 D1m-E91Q_1xG4S_bNAb1-HC Human CD4 mD1.22 + E91Q fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 124 D1m-E91H_1xG4S_bNAb1-HC Human CD4 mD1.22 + E91H fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 125 D1m-Q25E_1xG4S_bNAb1-HC Human CD4 mD1.22 + Q25E fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 126 D1m-H27D_1xG4S_bNAb1-HC Human CD4 mD1.22 + H27D fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 127 D1m-R58V_1xG4S_bNAb1-HC Human CD4 mD1.22 + R58V fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 128 D1m-R58N_1xG4S_bNAb1-HC Human CD4 mD1.22 + R58N fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 129 D1m-R58T_1xG4S_bNAb1-HC Human CD4 mD1.22 + R58T fused to the N- terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 130 D1m-L61M_1xG4S_bNAb1-HC Human CD4 mD1.22 + L61M fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 131 D1m-E13C-I70C_1xG4S_bNAb1-HC Human CD4 mD1.22 + E13C + I70C fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 132 D1m-H27C-G38C_1xG4S_bNAb1- Human CD4 mD1.22 + H27C + G38C fused HC to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 133 D1m-K21C-G65C_1xG4S_bNAb1- Human CD4 mD1.22 + K21C + G65C fused HC to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 134 D1m-E87G_1xG4S_bNAb1-HC Human CD4 mD1.22 + E87G fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 135 D1m-N52W_1xG4S_bNAb1-HC Human CD4 mD1.22 + N52W fused to the N-terminus of bNAb1 heavy chain with 1xG4S linker, Fc includes M428L/N434S 136 D1m-K8C-G99C_1xG4S_bNAb2-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb2 heavy chain with 1xG4S linker, Fc includes M428L/N434S 137 D1m-K8C-G99C_2xG4S_bNAb2-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb2 heavy chain with 2xG4S linker, Fc includes M428L/N434S 138 D1m-K8C-G99C_3xG4S_bNAb2-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb2 heavy chain with 3xG4S linker, Fc includes M428L/N434S 139 D1m-K8C-G99C_4xG4S_bNAb2-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb2 heavy chain with 4xG4S linker, Fc includes M428L/N434S 140 bNAb2-HC-mid_1xG4S_D1m-K8C- Human CD4 mD1.22 + K8C + G99C fused in G99C between the CH1 domain and hinge of bNAb2 heavy chain with 1xG4S linkers, Fc includes M428L/N434S 141 bNAb2-HC_1xG4S_D1m-K8C-G99C Human CD4 mD1.22 + K8C + G99C fused to the C-terminus of bNAb2 heavy chain with 1xG4S linker, Fc includes M428L/N434S 142 D1m-K8C-G99C_1xG4S_bNAb2-LC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb2 light chain with 1xG4S linker 143 bNAb2-LC_1xG4S_D1m-K8C-G99C Human CD4 mD1.22 + K8C + G99C fused to the C-terminus of bNAb2 light chain with 1xG4S linker 144 D1m-K8C-G99C_1xG4S_bNAb3-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb3 heavy chain with 1xG4S linker, Fc includes M428L/N434S 145 D1m-K8C-G99C_4xG4S_bNAb3-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb3 heavy chain with 4xG4S linker, Fc includes M428L/N434S 146 D1m-K8C-G99C_1xG4S_bNAb4-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb4 heavy chain with 1xG4S linker, Fc includes M428L/N434S 147 D1m-K8C-G99C_4xG4S_bNAb4-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb4 heavy chain with 4xG4S linker, Fc includes M428L/N434S 148 D1m-K8C-G99C_1xG4S_bNAb5-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb5 heavy chain with 1xG4S linker, Fc includes M428L/N434S 149 D1m-K8C-G99C_4xG4S_bNAb5-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb5 heavy chain with 4xG4S linker, Fc includes M428L/N434S 150 D1mD2_4xG4S_bNAb6-HC Human CD4 mD1.22-D2 fused to the N- terminus of bNAb6 heavy chain with 4xG4S linker, Fc includes M428L/N434S 151 D1m_4xG4S_bNAb6-HC Human CD4 mD1.22 fused to the N- terminus of bNAb6 heavy chain with 4xG4S linker, Fc includes M428L/N434S 152 D1m_4xG4S_bNAb1-scFv-H4Lss_2xG4S_Fc Human CD4 mD1.22 fused, via 4xG4S linker, to the N-terminus of bNAb1 scFv (VH-4xG4S-VL, with stabilizing disulfide bond G44C/HC-G100C/LC), which is fused, via 2xG4S linker, to the N-terminus of human Fc fragment including M428L/N434S 153 D1m_3xG4S_bNAb1-scFv-H4Lss_2xG4S_Fc Human CD4 mD1.22 fused, via 3xG4S linker, to the N-terminus of bNAb1 scFv (VH-4xG4S-VL, with stabilizing disulfide bond G44C/HC-G100C/LC), which is fused, via 2xG4S linker, to the N-terminus of human Fc fragment including M428L/N434S 154 D1m_2xG4S_bNAb1-scFv-H4Lss_2xG4S_Fc Human CD4 mD1.22 fused, via 2xG4S linker, to the N-terminus of bNAb1 scFv (VH-4xG4S-VL, with stabilizing disulfide bond G44C/HC-G100C/LC), which is fused, via 2xG4S linker, to the N-terminus of human Fc fragment including M428L/N434S 155 D1m_1xG4S_bNAb1-scFv-H4Lss_2xG4S_Fc Human CD4 mD1.22 fused, via 1xG4S linker, to the N-terminus of bNAb1 scFv (VH-4xG4S-VL, with stabilizing disulfide bond G44C/HC-G100C/LC), which is fused, via 2xG4S linker, to the N-terminus of human Fc fragment including M428L/N434S 156 Fc_4xG4S_D1m_3xG4S_bNAb1- Human CD4 mD1.22 fused, via 3xG4S scFv-H4Lss linker, to the N-terminus of bNAb1 scFv (VH-4xG4S-VL, with stabilizing disulfide bond G44C/HC-G100C/LC), which is fused, via 3xG4S linker, to the C-terminus of human Fc fragment including M428L/N434S 157 bNAb1-scFv-L4Hss_4xG4S_D1m_3xG4S_Fc bNAb1 scFv (VL-4xG4S-VH, with stabilizing disulfide bond G44C/HC-G100C/LC) fused, via 4xG4S linker, to the N-terminus of human CD4 mD1.22, which is fused, via 3xG4S linker, to the N-terminus of human Fc fragment including M428L/N434S 158 D1m-K8C-G99C_Fc Human CD4 mD1.22 + K8C, G99C with C- terminal Fc tag including M428L/N434S 159 bNAb7 CDRH1 CDRH1 of bNAb7 160 bNAb7 CDRH2 CDRH2 of bNAb7 161 bNAb7 CDRH3 CDRH3 of bNAb7 162 bNAb7 CDRL1 CDRL1 of bNAb7 163 bNAb7 CDRL2 CDRL2 of bNAb7 164 bNAb7 CDRL3 CDRL3 of bNAb7 165 bNAb8 CDRH1 CDRH1 of bNAb8 166 bNAb8 CDRH2 CDRH2 of bNAb8 167 bNAb8 CDRH3 CDRH3 of bNAb8 168 bNAb8 CDRL1 CDRL1 of bNAb8 169 bNAb8 CDRL2 CDRL2 of bNAb8 170 bNAb8 CDRL3 CDRL3 of bNAb8 171 bNAb9 CDRH1 CDRH1 of bNAb9 172 bNAb9 CDRH2 CDRH2 of bNAb9 173 bNAb9 CDRH3 CDRH3 of bNAb9 174 bNAb9 CDRL1 CDRL1 of bNAb9 175 bNAb9 CDRL2 CDRL2 of bNAb9 176 bNAb9 CDRL3 CDRL3 of bNAb9 177 bNAb10 CDRH1 CDRH1 of bNAb10 178 bNAb10 CDRH2 CDRH2 of bNAb10 179 bNAb10 CDRH3 CDRH3 of bNAb10 180 bNAb10 CDRL1 CDRL1 of bNAb10 181 bNAb10 CDRL2 CDRL2 of bNAb10 182 bNAb10 CDRL3 CDRL3 of bNAb10 183 bNAb11 CDRH1 CDRH1 of bNAb11 184 bNAb11 CDRH2 CDRH2 of bNAb11 185 bNAb11 CDRH3 CDRH3 of bNAb11 186 bNAb11 CDRL1 CDRL1 of bNAb11 187 bNAb11 CDRL2 CDRL2 of bNAb11 188 bNAb11 CDRL3 CDRL3 of bNAb11 189 bNAb12 CDRH1 CDRH1 of bNAb12 190 bNAb12 CDRH2 CDRH2 of bNAb12 191 bNAb12 CDRH3 CDRH3 of bNAb12 192 bNAb12 CDRL1 CDRL1 of bNAb12 193 bNAb12 CDRL2 CDRL2 of bNAb12 194 bNAb12 CDRL3 CDRL3 of bNAb12 195 bNAb13 CDRH1 CDRH1 of bNAb13 196 bNAb13 CDRH2 CDRH2 of bNAb13 197 bNAb13 CDRH3 CDRH3 of bNAb13 198 bNAb13 CDRL1 CDRL1 of bNAb13 199 bNAb13 CDRL2 CDRL2 of bNAb13 200 bNAb13 CDRL3 CDRL3 of bNAb13 201 bNAb14 CDRH1 CDRH1 of bNAb14 202 bNAb14 CDRH2 CDRH2 of bNAb14 203 bNAb14 CDRH3 CDRH3 of bNAb14 204 bNAb14 CDRL1 CDRL1 of bNAb14 205 bNAb14 CDRL2 CDRL2 of bNAb14 206 bNAb14 CDRL3 CDRL3 of bNAb14 207 bNAb15 CDRH1 CDRH1 of bNAb15 208 bNAb15 CDRH2 CDRH2 of bNAb15 209 bNAb15 CDRH3 CDRH3 of bNAb15 210 bNAb15 CDRL1 CDRL1 of bNAb15 211 bNAb15 CDRL2 CDRL2 of bNAb15 212 bNAb15 CDRL3 CDRL3 of bNAb15 213 bNAb16 CDRH1 CDRH1 of bNAb16 214 bNAb16 CDRH2 CDRH2 of bNAb16 215 bNAb16 CDRH3 CDRH3 of bNAb16 216 bNAb16 CDRL1 CDRL1 of bNAb16 217 bNAb16 CDRL2 CDRL2 of bNAb16 218 bNAb16 CDRL3 CDRL3 of bNAb16 219 bNAb17 CDRH1 CDRH1 of bNAb17 220 bNAb17 CDRH2 CDRH2 of bNAb17 212 bNAb17 CDRH3 CDRH3 of bNAb17 222 bNAb17 CDRL1 CDRL1 of bNAb17 223 bNAb17 CDRL2 CDRL2 of bNAb17 224 bNAb17 CDRL3 CDRL3 of bNAb17 225 bNAb18 CDRH1 CDRH1 of bNAb18 226 bNAb18 CDRH2 CDRH2 of bNAb18 227 bNAb18 CDRH3 CDRH3 of bNAb18 228 bNAb18 CDRL1 CDRL1 of bNAb18 229 bNAb18 CDRL2 CDRL2 of bNAb18 230 bNAb18 CDRL3 CDRL3 of bNAb18 231 bNAb19 CDRH1 CDRH1 of bNAb19 232 bNAb19 CDRH2 CDRH2 of bNAb19 233 bNAb19 CDRH3 CDRH3 of bNAb19 234 bNAb19 CDRL1 CDRL1 of bNAb19 235 bNAb19 CDRL2 CDRL2 of bNAb19 236 bNAb19 CDRL3 CDRL3 of bNAb19 237 bNAb20 CDRH1 CDRH1 of bNAb20 238 bNAb20 CDRH2 CDRH2 of bNAb20 239 bNAb20 CDRH3 CDRH3 of bNAb20 240 bNAb20 CDRL1 CDRL1 of bNAb20 241 bNAb20 CDRL2 CDRL2 of bNAb20 242 bNAb20 CDRL3 CDRL3 of bNAb20 243 bNAb21 and bNAb22 CDRH1 CDRH1 of bNAb21 and bNAb22 244 bNAb21 and bNAb22 CDRH2 CDRH2 of bNAb21 and bNAb22 245 bNAb21 and bNAb22 CDRH3 CDRH3 of bNAb21 and bNAb22 246 bNAb21 and bNAb22 CDRL1 CDRL1 of bNAb21 and bNAb22 247 bNAb21 and bNAb22 CDRL2 CDRL2 of bNAb21 and bNAb22 248 bNAb21 and bNAb22 CDRL3 CDRL3 of bNAb21 and bNAb22 249 bNAb23 CDRH1 CDRH1 of bNAb23 250 bNAb23 CDRH2 CDRH2 of bNAb23 251 bNAb23 CDRH3 CDRH3 of bNAb23 252 bNAb23 CDRL1 CDRL1 of bNAb23 253 bNAb23 CDRL2 CDRL2 of bNAb23 254 bNAb23 CDRL3 CDRL3 of bNAb23 255 bNAb7 VH Heavy chain variable region of bNAb7 256 bNAb7 VL Light chain variable region of bNAb7 257 bNAb7 HC Full heavy chain of bNAb7 258 bNAb7 HC + LS Full heavy chain of bNAb7 with M428L/N434S mutations 259 bNAb7 LC Full light chain of bNAb7 260 bNAb8 VH Heavy chain variable region of bNAb8 261 bNAb8 VL Light chain variable region of bNAb8 262 bNAb8 HC Full heavy chain of bNAb8 263 bNAb8 HC + LS Full heavy chain of bNAb8 with M428L/N434S mutations 264 bNAb8 LC Full light chain of bNAb8 265 bNAb9 VH Heavy chain variable region of bNAb9 266 bNAb9 VL Light chain variable region of bNAb9 267 bNAb9 HC Full heavy chain of bNAb9 268 bNAb9 HC + LS Full heavy chain of bNAb9 with M428L/N434S mutations 269 bNAb9 LC Full light chain of bNAb9 270 bNAb10 VH Heavy chain variable region of bNAb10 271 bNAb10 VL Light chain variable region of bNAb10 272 bNAb10 HC Full heavy chain of bNAb10 273 bNAb10 HC + LS Full heavy chain of bNAb10 with M428L/N434S mutations 274 bNAb10 LC Full light chain of bNAb10 275 bNAb11 VH Heavy chain variable region of bNAb11 276 bNAb11 VL Light chain variable region of bNAb11 277 bNAb11 HC Full heavy chain of bNAb11 278 bNAb11 HC + LS Full heavy chain of bNAb11 with M428L/N434S mutations 279 bNAb11 LC Full light chain of bNAb11 280 bNAb12 VH Heavy chain variable region of bNAb12 281 bNAb12 VL Light chain variable region of bNAb12 282 bNAb12 HC Full heavy chain of bNAb12 283 bNAb12 HC + LS Full heavy chain of bNAb12 with M428L/N434S mutations 284 bNAb12 LC Full light chain of bNAb12 285 bNAb13 VH Heavy chain variable region of bNAb13 286 bNAb13 VL Light chain variable region of bNAb13 287 bNAb13 HC Full heavy chain of bNAb13 288 bNAb13 HC + LS Full heavy chain of bNAb13 with M428L/N434S mutations 289 bNAb13 LC Full light chain of bNAb13 290 bNAb14 VH Heavy chain variable region of bNAb13 291 bNAb14 VL Light chain variable region of bNAb14 292 bNAb14 HC Full heavy chain of bNAb14 293 bNAb14 HC + LS Full heavy chain of bNAb14 with M428L/N434S mutations 294 bNAb14 LC Full light chain of bNAb14 295 bNAb15 VH Heavy chain variable region of bNAb15 296 bNAb15 VL Light chain variable region of bNAb15 297 bNAb15 HC Full heavy chain of bNAb15 298 bNAb15 HC + LS Full heavy chain of bNAb15 with M428L/N434S mutations 299 bNAb15 LC Full light chain of bNAb15 300 bNAb16 VH Heavy chain variable region of bNAb16 301 bNAb16 VL Light chain variable region of bNAb16 302 bNAb16 HC Full heavy chain of bNAb16 303 bNAb16 HC + LS Full heavy chain of bNAb16 with M428L/N434S mutations 304 bNAb16 LC Full light chain of bNAb16 305 bNAb17 VH Heavy chain variable region of bNAb17 306 bNAb17 VL Light chain variable region of bNAb17 307 bNAb17 HC Full heavy chain of bNAb17 308 bNAb17 HC + LS Full heavy chain of bNAb17 with M428L/N434S mutations 309 bNAb17 LC Full light chain of bNAb17 310 bNAb18 VH Heavy chain variable region of bNAb17 311 bNAb18 VL Light chain variable region of bNAb17 312 bNAb18 HC Full heavy chain of bNAb17 313 bNAb18 HC + LS Full heavy chain of bNAb17 with M428L/N434S mutations 314 bNAb18 LC Full light chain of bNAb18 315 bNAb19 VH Heavy chain variable region of bNAb19 316 bNAb19 VL Light chain variable region of bNAb19 317 bNAb19 HC Full heavy chain of bNAb19 318 bNAb19 HC + LS Full heavy chain of bNAb19 with M428L/N434S mutations 319 bNAb19 LC Full light chain of bNAb19 320 bNAb20 VH Heavy chain variable region of bNAb20 321 bNAb20 VL Light chain variable region of bNAb20 322 bNAb20 HC Full heavy chain of bNAb20 323 bNAb20 HC + LS Full heavy chain of bNAb20 with M428L/N434S mutations 324 bNAb20 LC Full light chain of bNAb20 325 bNAb21 VH Heavy chain variable region of bNAb21 326 bNAb21 VL Light chain variable region of bNAb21 327 bNAb21 HC Full heavy chain of bNAb21 328 bNAb21 HC + LS Full heavy chain of bNAb21 with M428L/N434S mutations 329 bNAb21 LC Full light chain of bNAb21 330 bNAb22 VH Full heavy chain of bNAb22 331 bNAb22 HC Full heavy chain of bNAb22 with M428L/N434S mutations 332 bNAb22 HV + LS Full light chain of bNAb22 333 bNAb23 VH Heavy chain variable region of bNAb23 334 bNAb23 VL Light chain variable region of bNAb23 335 bNAb23 HC Full heavy chain of bNAb23 336 bNAb23 HC + LS Full heavy chain of bNAb23 with M428L/N434S mutations 337 bNAb23 LC Full light chain of bNAb23 338 D1m-K8C-G99C_1xG4S_bNAb7-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb7 heavy chain with 1xG4S linker, Fc includes M428L/N434S 339 D1m-K8C-G99C_1xG4S_bNAb8-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb8 heavy chain with 1xG4S linker, Fc includes M428L/N434S 340 D1m-K8C-G99C_1xG4S_bNAb9-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb9 heavy chain with 1xG4S linker, Fc includes M428L/N434S 341 D1m-K8C-G99C_1xG4S_bNAb10- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb10 heavy chain with 1xG4S linker, Fc includes M428L/N434S 342 D1m-K8C-G99C_1xG4S_bNAb11- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb11 heavy chain with 1xG4S linker, Fc includes M428L/N434S 343 D1m-K8C-G99C_1xG4S_bNAb12- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb12 heavy chain with 1xG4S linker, Fc includes M428L/N434S 344 bNAb12_HC_1xG4S_D1m-K8C- Human CD4 mD1.22 + K8C + G99C fused G99C to the C-terminus of bNAb12 heavy chain with 1xG4S linker, Fc includes M428L/N434S 345 D1m-K8C-G99C_1xG4S_LC-bNAb12 Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb12 light chain with 1xG4S linker 346 bNAb12_LC_1xG4S_D1m-K8C- Human CD4 mD1.22 + K8C + G99C fused G99C-HC to the C-terminus of bNAb12 light chain with 1xG4S linker 347 D1m-K8C-G99C_1xG4S_bNAb13- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb13 heavy chain with 1xG4S linker, Fc includes M428L/N434S 348 D1m-K8C-G99C_1xG4S_bNAb14- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb14 heavy chain with 1xG4S linker, Fc includes M428L/N434S 349 D1m-K8C-G99C_1xG4S_bNAb15- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb15 heavy chain with 1xG4S linker, Fc includes M428L/N434S 350 D1m-K8C-G99C_1xG4S_bNAb16- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb16 heavy chain with 1xG4S linker, Fc includes M428L/N434S 351 D1m-K8C-G99C_1xG4S_bNAb17- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb17 heavy chain with 1xG4S linker, Fc includes M428L/N434S 352 D1m-K8C-G99C_1xG4S_bNAb18- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb18 heavy chain with 1xG4S linker, Fc includes M428L/N434S 353 D1m-K8C-G99C_1xG4S_bNAb19- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb19 heavy chain with 1xG4S linker, Fc includes M428L/N434S 354 D1m-K8C-G99C_1xG4S_bNAb20- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb20 heavy chain with 1xG4S linker, Fc includes M428L/N434S 355 D1m-K8C-G99C_1xG4S_bNAb21- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb21 heavy chain with 1xG4S linker, Fc includes M428L/N434S 356 D1m-K8C-G99C_1xG4S_bNAb22- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb22 heavy chain with 1xG4S linker, Fc includes M428L/N434S 357 D1m-K8C-G99C_1xG4S_bNAb23- Human CD4 mD1.22 + K8C + G99C fused HC to the N-terminus of bNAb23 heavy chain with 1xG4S linker, Fc includes M428L/N434S 358 D1m-K8C-G99C_1xG4S_bNAb23-LC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb23 light chain with 1xG4S linker 359 bNAb23-HC_1xG4S_D1m-K8C- Human CD4 mD1.22 + K8C + G99C fused G99C to the C-terminus of bNAb23 heavy chain with 1xG4S linker, Fc includes M428L/N434S 360 bNAb23_LC_1xG4S_D1m-K8C- Human CD4 mD1.22 + K8C + G99C fused G99C to the C-terminus of bNAb23 light chain with 1xG4S linker 361 V3 loop V3 loop consensus sequence 362 D1m-K8C-G99C_1xG4S_bNAb6-HC Human CD4 mD1.22 + K8C + G99C fused to the N-terminus of bNAb6 heavy chain with 1xG4S linker, Fc includes M428L/N434S 363 Exemplary gp160 Exemplary gp160 sequence 364 Exemplary gp120 Exemplary gp120 sequence 365 bNAb22 VL Light chain variable region of bNAb22 366 bNAb22 LC Full light chain of bNAb22
(318) TABLE-US-00034 SEQIDNO:1 KKVVLGKKGDTVELTCTASQKKSIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLA SEQIDNO:2 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAF SEQIDNO:3 KKVVLGKKGDTVELTCTASQKKSIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLLVFG SEQIDNO:4 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:5 KKVVYGKVGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:6 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK QEVQLVVVG SEQIDNO:7 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK HEVQLVVVG SEQIDNO:8 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVGDQK EEVQLVVVG SEQIDNO:9 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLWDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:10 KKVVYGKIGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:11 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVC SEQIDNO:12 KKVVYGKKGDCVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIICNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:13 KKVVYGKKGDTVCLTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLCIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:14 KKVVYGKKGDTVELTCTASQKKNIQFCWKNSNQIKILCNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:15 KKVVYGKKGDTVELTCTASQCKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQCNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:16 KKVVYGKKGDTVELTCTASQKKNIEFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:17 KKVVYGKKGDTVELTCTASQKKNIQFDWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:18 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSVRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:19 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSNRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:20 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSTRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:21 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSMWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVG SEQIDNO:22 ACNSFWG SEQIDNO:23 SLSHCASYWNRGWTYHNPSLKS SEQIDNO:24 FGGEVLRYTDWPKPAWVDL SEQIDNO:25 TGTSNNFVS SEQIDNO:26 DVNKRPS SEQIDNO:27 GSLVGNWDVI SEQIDNO:28 DSYWS SEQIDNO:29 YVHKSGDTNYSPSLKS SEQIDNO:30 TLHGRRIYGIVAFNEWFTYFYMDV SEQIDNO:31 GEKSLGSRAVQ SEQIDNO:32 NNQDRPS SEQIDNO:33 HIWDSRVPTKWV SEQIDNO:34 SDHSWT SEQIDNO:35 DIHYNGATTYNPSLRS SEQIDNO:36 NAIRIYGVVALGEWFHYGMDV SEQIDNO:37 SGAPLTSRFTY SEQIDNO:38 RSSQRSS SEQIDNO:39 QSSDTSDSYKM SEQIDNO:40 NYYWT SEQIDNO:41 YISDRESATYNPSLNS SEQIDNO:42 ARRGQRIYGVVSFGEFFYYYSMDV SEQIDNO:43 GRQALGSRAVQ SEQIDNO:44 NNQDRPS SEQIDNO:45 HMWDSRSGFSWS SEQIDNO:46 GGEWGDKDYHWG SEQIDNO:47 SIHWRGTTHYKESLRR SEQIDNO:48 HRHHDVFMLVPIAGWFDV SEQIDNO:49 RASQNINKNLA SEQIDNO:50 ETYSKIA SEQIDNO:51 QQYEEWPRT SEQIDNO:52 DFYIH SEQIDNO:53 WMNPQTGRTNTARNFQG SEQIDNO:54 GGWISLYYDSSYYPNFDH SEQIDNO:55 TGTKYDVGSHDLVS SEQIDNO:56 EVNKRPS SEQIDNO:57 CSFGGSATVV SEQIDNO:58 QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGK GLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKL NSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSS SEQIDNO:59 QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVI YDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVG NWDVIFGGGTKLTVL SEQIDNO:60 QSALTQPPSASGSPGQSITISCTGTSNNYVSWYQQHAGKAPKLVI YDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVG NWDVIFGGGTKLTVL SEQIDNO:61 QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGK GLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKL NSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK SEQIDNO:62 QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGK GLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKL NSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSWVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK SEQIDNO:63 QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVI YDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVG NWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:64 QSALTQPPSASGSPGQSITISCTGTSNNYVSWYQQHAGKAPKLVI YDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVG NWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:65 QMQLQESGPGLVKPSETLSLTCSVSGASISDSYWSWIRRSPGKGL EWIGYVHKSGDTNYSPSLKSRVNLSLDTSKNQVSLSLVAATAADS GKYYCARTLHGRRIYGIVAFNEWFTYFYMDVWGNGTQVTVSS SEQIDNO:66 SDISVAPGETARISCGEKSLGSRAVQWYQHRAGQAPSLIIYNNQD RPSGIPERFSGSPDSPFGTTATLTITSVEAGDEADYYCHIWDSRV PTKWVFGGGTTLTVL SEQIDNO:67 QMQLQESGPGLVKPSETLSLTCSVSGASISDSYWSWIRRSPGKGL EWIGYVHKSGDTNYSPSLKSRVNLSLDTSKNQVSLSLVAATAADS GKYYCARTLHGRRIYGIVAFNEWFTYFYMDVWGNGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:68 QMQLQESGPGLVKPSETLSLTCSVSGASISDSYWSWIRRSPGKGL EWIGYVHKSGDTNYSPSLKSRVNLSLDTSKNQVSLSLVAATAADS GKYYCARTLHGRRIYGIVAFNEWFTYFYMDVWGNGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:69 SDISVAPGETARISCGEKSLGSRAVQWYQHRAGQAPSLIIYNNQD RPSGIPERFSGSPDSPFGTTATLTITSVEAGDEADYYCHIWDSRV PTKWVFGGGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHKSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:70 QVQLRESGPGLVKPSETLSLSCTVSNDSRPSDHSWTWVRQSPGKA LEWIGDIHYNGATTYNPSLRSRVRIELDQSIPRFSLKMTSMTAAD TGMYYCARNAIRIYGVVALGEWFHYGMDVWGQGTAVTVSS SEQIDNO:71 SSELTQPPSVSVSPGQTARITCSGAPLTSRFTYWYRQKPGQAPVL IISRSSQRSSGWSGRFSASWSGTTVTLTIRGVQADDEADYYCQSS DTSDSYKMFGGGTKLTVL SEQIDNO:72 QVQLRESGPGLVKPSETLSLSCTVSNDSRPSDHSWTWVRQSPGKA LEWIGDIHYNGATTYNPSLRSRVRIELDQSIPRFSLKMTSMTAAD TGMYYCARNAIRIYGVVALGEWFHYGMDVWGQGTAVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK SEQIDNO:73 QVQLRESGPGLVKPSETLSLSCTVSNDSRPSDHSWTWVRQSPGKA LEWIGDIHYNGATTYNPSLRSRVRIELDQSIPRFSLKMTSMTAAD TGMYYCARNAIRIYGVVALGEWFHYGMDVWGQGTAVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYT QKSLSLSPGK SEQIDNO:74 SSELTQPPSVSVSPGQTARITCSGAPLTSRFTYWYRQKPGQAPVL IISRSSQRSSGWSGRFSASWSGTTVTLTIRGVQADDEADYYCQSS DTSDSYKMFGGGTKLTVLGQPAAAPSVTLFPPSSEELQANKATLV CLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYL SLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTEC SEQIDNO:75 QVQLQESGPGLVKPSETLSVTCSVSGDSMNNYYWTWIRQSPGKGL EWIGYISDRESATYNPSLNSRVVISRDTSKNQLSLKLNSVTPADT AVYYCATARRGQRIYGVVSFGEFFYYYSMDVWGKGTTVTVSS SEQIDNO:76 SYVRPLSVALGETARISCGRQALGSRAVQWYQHRPGQAPILLIYN NQDRPSGIPERFSGTPDINFGTRATLTISGVEAGDEADYYCHMWD SRSGFSWSFGGATRLTVL SEQIDNO:77 QVQLQESGPGLVKPSETLSVTCSVSGDSMNNYYWTWIRQSPGKGL EWIGYISDRESATYNPSLNSRVVISRDTSKNQLSLKLNSVTPADT AVYYCATARRGQRIYGVVSFGEFFYYYSMDVWGKGTTVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:78 QVQLQESGPGLVKPSETLSVTCSVSGDSMNNYYWTWIRQSPGKGL EWIGYISDRESATYNPSLNSRVVISRDTSKNQLSLKLNSVTPADT AVYYCATARRGQRIYGVVSFGEFFYYYSMDVWGKGTTVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:79 SYVRPLSVALGETARISCGRQALGSRAVQWYQHRPGQAPILLIYN NQDRPSGIPERFSGTPDINFGTRATLTISGVEAGDEADYYCHMWD SRSGFSWSFGGATRLTVLGQPKAAPSVTLFPPSSEELQANKATLV CLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYL SLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:80 QLQMQESGPGLVKPSETLSLSCTVSGDSIRGGEWGDKDYHWGWVR HSAGKGLEWIGSIHWRGTTHYKESLRRRVSMSIDTSRNWFSLRLA SVTAADTAVYFCARHRHHDVFMLVPIAGWFDVWGPGVQVTVSS SEQIDNO:81 EIVMTQSPDTLSVSPGETVTLSCRASQNINKNLAWYQYKPGQSPR LVIFETYSKIAAFPARFVASGSGTEFTLTINNMQSEDVAVYYCQQ YEEWPRTFGQGTKVDIK SEQIDNO:82 QLQMQESGPGLVKPSETLSLSCTVSGDSIRGGEWGDKDYHWGWVR HSAGKGLEWIGSIHWRGTTHYKESLRRRVSMSIDTSRNWFSLRLA SVTAADTAVYFCARHRHHDVFMLVPIAGWFDVWGPGVQVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK SEQIDNO:83 QLQMQESGPGLVKPSETLSLSCTVSGDSIRGGEWGDKDYHWGWVR HSAGKGLEWIGSIHWRGTTHYKESLRRRVSMSIDTSRNWFSLRLA SVTAADTAVYFCARHRHHDVFMLVPIAGWFDVWGPGVQVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGK SEQIDNO:84 EIVMTQSPDTLSVSPGETVTLSCRASQNINKNLAWYQYKPGQSPR LVIFETYSKIAAFPARFVASGSGTEFTLTINNMQSEDVAVYYCQQ YEEWPRTFGQGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:85 QVQLVQSGAQMKNPGASVKVSCAPSGYTFTDFYIHWLRQAPGQGL QWMGWMNPQTGRTNTARNFQGRVTMTRDTSIGTAYMELRSLTSDD TAIYYCTTGGWISLYYDSSYYPNFDHWGQGTLLTVSS SEQIDNO:86 QSALTQPASVSGSPGQSITISCTGTKYDVGSHDLVSWYQQYPGKV PKYMIYEVNKRPSGVSNRFSGSKSGNTASLTISGLRAEDEADYYC CSFGGSATVVCGGGTKVTVL SEQIDNO:87 QVQLVQSGAQMKNPGASVKVSCAPSGYTFTDFYIHWLRQAPGQGL QWMGWMNPQTGRTNTARNFQGRVTMTRDTSIGTAYMELRSLTSDD TAIYYCTTGGWISLYYDSSYYPNFDHWGQGTLLTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT CVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK SEQIDNO:88 QVQLVQSGAQMKNPGASVKVSCAPSGYTFTDFYIHWLRQAPGQGL QWMGWMNPQTGRTNTARNFQGRVTMTRDTSIGTAYMELRSLTSDD TAIYYCTTGGWISLYYDSSYYPNFDHWGQGTLLTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKS LSLSPGK SEQIDNO:89 QSALTQPASVSGSPGQSITISCTGTKYDVGSHDLVSWYQQYPGKV PKYMIYEVNKRPSGVSNRFSGSKSGNTASLTISGLRAEDEADYYC CSFGGSATVVCGGGTKVTVLGQPKAAPSVTLFPPSSEELQANKAT LVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:90 GGGGS SEQIDNO:91 GGGGSGGGGS SEQIDNO:92 GGGGSGGGGSGGGGS SEQIDNO:93 GGGGSGGGGSGGGGSGGGGS SEQIDNO:94 GGGGSGGGGSGGGGSGGGGSGGGGS SEQIDNO:95 GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS SEQIDNO:96 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFQ PQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGKG LEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKLN SVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALH SHYTQKSLSLSPGK SEQIDNO:97 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFG GGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQ PPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLV FLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPRE EQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHSHYTQKSLSLSPGK SEQIDNO:98 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFG GGGSGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFW GWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDT PKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWG RGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:99 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFG GGGSGGGGSGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:100 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFG GGGSGGGGSGGGGSGGGGSQPQLQESGPTLVEASETLSLTCAVSG DSTAACNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSL KSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDW PKPAWVDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:101 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGQPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFW GWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDT PKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWG RGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:102 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:103 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSQPQLQESGPTLVEASETLSLTCAVSG DSTAACNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSL KSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDW PKPAWVDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALG CLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:104 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSQPQLQESGPTLVEASETLSLT CAVSGDSTAACNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTY HNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVL RYTDWPKPAWVDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:105 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSQPQLQESGPTLVEASE TLSLTCAVSGDSTAACNSFWGWVRQPPGKGLEWVGSLSHCASYWN RGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATYYCARF GGEVLRYTDWPKPAWVDLWGRGTLVTVSSASTKGPSVFPLAPSSK STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEL TKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:106 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSGGGGSQPQLQESGPTL VEASETLSLTCAVSGDSTAACNSFWGWVRQPPGKGLEWVGSLSHC ASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATY YCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPELLGGPSVFLEPPKPKDTLMISRTPEVTCVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLS PGK SEQIDNO:107 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSQPQLQE SGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGKGLEWVG SLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAA DTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQ KSLSLSPGK SEQIDNO:108 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGQSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQH AGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEA VYYCGSLVGNWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQA NKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKY AASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:109 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQSALTQPPSASGSPGQSITISCTGTSNNFVS WYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTASLTVSGLQ TDDEAVYYCGSLVGNWDVIFGGGTKLTVLGQPKAAPSVTLFPPSS EELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQ SNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:110 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSQSALTQPPSASGSPGQSITISCTGTS NNFVSWYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTASLT VSGLQTDDEAVYYCGSLVGNWDVIFGGGTKLTVLGQPKAAPSVTL FPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETT TPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVA PTECS SEQIDNO:111 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSQSALTQPPSASGSPGQSITIS CTGTSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGN TASLTVSGLQTDDEAVYYCGSLVGNWDVIFGGGTKLTVLGQPKAA PSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKA GVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTV EKTVAPTECS SEQIDNO:112 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSQSALTQPPSASGSPGQ SITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVPDRFSG SKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIFGGGTKLTVLG QPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADS SPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS SEQIDNO:113 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSGGGGSQSALTQPPSAS GSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVP DRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIFGGGTK LTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVA WKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYS CQVTHEGSTVEKTVAPTECS SEQIDNO:114 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSQSALTQ PPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKR PSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIF GGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPG AVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:115 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQSALTQPPSASGSPGQSITISCTGTSNNFVS WYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTASLTVSGLQ TDDEAVYYCGSLVGNWDVIFGGGTKLTVLGQPKAAPSVTLFPPSS EELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQ SNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:116 QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGK GLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKL NSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCGGGGSKKVVYGKCGDTVELTCTASQKKNIQFH WKNSNQIKILGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFPLIIK NLKPEDSDTYICEVEDQKEEVQLVVVCGGGGSDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:117 QPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGK GLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKL NSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGGGGGSKKVVYGKCGDTVELTCTASQKKNIQF HWKNSNQIKILGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFPLII KNLKPEDSDTYICEVEDQKEEVQLVVVC SEQIDNO:118 QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVI YDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVG NWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECSGGGGSKKVVYGKCG DTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLTKGPSKLNDR VDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQKEEVQLVVVC SEQIDNO:119 KKVVYGKIGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:120 KKVVYGKVGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:121 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:122 KKVVYGKKGDCVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIICNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:123 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK QEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:124 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK HEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:125 KKVVYGKKGDTVELTCTASQKKNIEFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:126 KKVVYGKKGDTVELTCTASQKKNIQFDWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:127 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSVRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:128 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSNRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:129 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSTRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:130 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSMWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:131 KKVVYGKKGDTVCLTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLCIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:132 KKVVYGKKGDTVELTCTASQKKNIQFCWKNSNQIKILCNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:133 KKVVYGKKGDTVELTCTASQCKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQCNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:134 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVGDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:135 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLWDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKGLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:136 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQMQLQESGPGLVKPSETLSLTCSVSGASISD SYWSWIRRSPGKGLEWIGYVHKSGDTNYSPSLKSRVNLSLDTSKN QVSLSLVAATAADSGKYYCARTLHGRRIYGIVAFNEWFTYFYMDV WGNGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:137 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSGGGGSQMQLQESGPGLVKPSETLSLTCSVSG ASISDSYWSWIRRSPGKGLEWIGYVHKSGDTNYSPSLKSRVNLSL DTSKNQVSLSLVAATAADSGKYYCARTLHGRRIYGIVAFNEWFTY FYMDVWGNGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:138 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSGGGGSGGGGSQMQLQESGPGLVKPSETLSLT CSVSGASISDSYWSWIRRSPGKGLEWIGYVHKSGDTNYSPSLKSR VNLSLDTSKNQVSLSLVAATAADSGKYYCARTLHGRRIYGIVAFN EWFTYFYMDVWGNGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:139 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSGGGGSGGGGSGGGGSQMQLQESGPGLVKPSE TLSLTCSVSGASISDSYWSWIRRSPGKGLEWIGYVHKSGDTNYSP SLKSRVNLSLDTSKNQVSLSLVAATAADSGKYYCARTLHGRRIYG IVAFNEWFTYFYMDVWGNGTQVTVSSASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:140 QMQLQESGPGLVKPSETLSLTCSVSGASISDSYWSWIRRSPGKGL EWIGYVHKSGDTNYSPSLKSRVNLSLDTSKNQVSLSLVAATAADS GKYYCARTLHGRRIYGIVAFNEWFTYFYMDVWGNGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCGGGGSKKVVYGKCGDTVELTCTASQKKNIQFHWKN SNQIKILGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFPLIIKNLK PEDSDTYICEVEDQKEEVQLVVVCGGGGSDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:141 QMQLQESGPGLVKPSETLSLTCSVSGASISDSYWSWIRRSPGKGL EWIGYVHKSGDTNYSPSLKSRVNLSLDTSKNQVSLSLVAATAADS GKYYCARTLHGRRIYGIVAFNEWFTYFYMDVWGNGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGGGGGSKKVVYGKCGDTVELTCTASQKKNIQFHWK NSNQIKILGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFPLIIKNL KPEDSDTYICEVEDQKEEVQLVVVC SEQIDNO:142 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSSDISVAPGETARISCGEKSLGSRAVQWYQHR AGQAPSLIIYNNQDRPSGIPERFSGSPDSPFGTTATLTITSVEAG DEADYYCHIWDSRVPTKWVFGGGTTLTVLGQPKAAPSVTLFPPSS EELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQ SNNKYAASSYLSLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:143 SDISVAPGETARISCGEKSLGSRAVQWYQHRAGQAPSLIIYNNQD RPSGIPERFSGSPDSPFGTTATLTITSVEAGDEADYYCHIWDSRV PTKWVFGGGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHKSYSCQVTHEGSTVEKTVAPTECSGGGGSKKVVYGKCG DTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLTKGPSKLNDR VDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQKEEVQLVVVC SEQIDNO:144 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLRESGPGLVKPSETLSLSCTVSNDSRPS DHSWTWVRQSPGKALEWIGDIHYNGATTYNPSLRSRVRIELDQSI PRFSLKMTSMTAADTGMYYCARNAIRIYGVVALGEWFHYGMDVWG QGTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:145 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSGGGGSGGGGSGGGGSQVQLRESGPGLVKPSE TLSLSCTVSNDSRPSDHSWTWVRQSPGKALEWIGDIHYNGATTYN PSLRSRVRIELDQSIPRFSLKMTSMTAADTGMYYCARNAIRIYGV VALGEWFHYGMDVWGQGTAVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SWVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:146 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLQESGPGLVKPSETLSVTCSVSGDSMNN YYWTWIRQSPGKGLEWIGYISDRESATYNPSLNSRVVISRDTSKN QLSLKLNSVTPADTAVYYCATARRGQRIYGVVSFGEFFYYYSMDV WGKGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:147 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGGGGGSGGGGSGGGGSQVQLQESGPGLVKPSET LSVTCSVSGDSMNNYYWTWIRQSPGKGLEWIGYISDRESATYNPS LNSRVVISRDTSKNQLSLKLNSVTPADTAVYYCATARRGQRIYGV VSFGEFFYYYSMDVWGKGTTVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:148 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQLQMQESGPGLVKPSETLSLSCTVSGDSIRG GEWGDKDYHWGWVRHSAGKGLEWIGSIHWRGTTHYKESLRRRVSM SIDTSRNWFSLRLASVTAADTAVYFCARHRHHDVFMLVPIAGWFD VWGPGVQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:149 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSGGGGSGGGGSGGGGSQLQMQESGPGLVKPSE TLSLSCTVSGDSIRGGEWGDKDYHWGWVRHSAGKGLEWIGSIHWR GTTHYKESLRRRVSMSIDTSRNWFSLRLASVTAADTAVYFCARHR HHDVFMLVPIAGWFDVWGPGVQVTVSSASTKGPSVFPLAPSSKST SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSWVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:150 KKVVLGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQK EEVQLVVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRG KNIQGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFG GGGSGGGGSGGGGSGGGGSQVQLVQSGAQMKNPGASVKVSCAPSG YTFTDFYIHWLRQAPGQGLQWMGWMNPQTGRTNTARNFQGRVTMT RDTSIGTAYMELRSLTSDDTAIYYCTTGGWISLYYDSSYYPNFDH WGQGTLLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:151 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSQVQLVQSGAQMKNPGA SVKVSCAPSGYTFTDFYIHWLRQAPGQGLQWMGWMNPQTGRTNTA RNFQGRVTMTRDTSIGTAYMELRSLTSDDTAIYYCTTGGWISLYY DSSYYPNFDHWGQGTLLTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWV TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:152 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSGGGGSQPQLQESGPTLVEASE TLSLTCAVSGDSTAACNSFWGWVRQPPGKCLEWVGSLSHCASYWN RGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATYYCARF GGEVLRYTDWPKPAWVDLWGRGTLVTVSSGGGGSGGGGSGGGGSG GGGSQSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAP KLVIYDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCG SLVGNWDVIFGCGTKLTVLGGGGSGGGGSDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:153 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSGGGGSQPQLQESGPTLVEASETLSLT CAVSGDSTAACNSFWGWVRQPPGKCLEWVGSLSHCASYWNRGWTY HNPSLKSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVL RYTDWPKPAWVDLWGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSQ SALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIY DVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGN WDVIFGCGTKLTVLGGGGSGGGGSDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:154 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSGGGGSQPQLQESGPTLVEASETLSLTCAVSG DSTAACNSFWGWVRQPPGKCLEWVGSLSHCASYWNRGWTYHNPSL KSRLTLALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDW PKPAWVDLWGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSQSALTQ PPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKR PSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIF GCGTKLTVLGGGGSGGGGSDKTHTCPPCPAPELLGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VLHEALHSHYTQKSLSLSPGK SEQIDNO:155 KKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVGGGGGSQPQLQESGPTLVEASETLSLTCAVSGDSTAA CNSFWGWVRQPPGKCLEWVGSLSHCASYWNRGWTYHNPSLKSRLT LALDTPKNLVFLKLNSVTAADTATYYCARFGGEVLRYTDWPKPAW VDLWGRGTLVTVSSGGGGSGGGGSGGGGSGGGGSQSALTQPPSAS GSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVP DRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVGNWDVIFGCGTK LTVLGGGGSGGGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEA LHSHYTQKSLSLSPGK SEQIDNO:156 DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR DELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSP GKGGGGSGGGGSGGGGSGGGGSKKVVYGKKGDTVELTCTASQKKN IQFHWKNSNQIKILGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFP LIIKNLKPEDSDTYICEVEDQKEEVQLVVVGGGGGSGGGGSGGGG SQPQLQESGPTLVEASETLSLTCAVSGDSTAACNSFWGWVRQPPG KCLEWVGSLSHCASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLK LNSVTAADTATYYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVS SGGGGSGGGGSGGGGSGGGGSQSALTQPPSASGSPGQSITISCTG TSNNFVSWYQQHAGKAPKLVIYDVNKRPSGVPDRFSGSKSGNTAS LTVSGLQTDDEAVYYCGSLVGNWDVIFGCGTKLTVL SEQIDNO:157 QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQHAGKAPKLVI YDVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCGSLVG NWDVIFGCGTKLTVLGGGGSGGGGSGGGGSGGGGSQPQLQESGPT LVEASETLSLTCAVSGDSTAACNSFWGWVRQPPGKCLEWVGSLSH CASYWNRGWTYHNPSLKSRLTLALDTPKNLVFLKLNSVTAADTAT YYCARFGGEVLRYTDWPKPAWVDLWGRGTLVTVSSGGGGGGGGSG GGGSGGGGSKKVVYGKKGDTVELTCTASQKKNIQFHWKNSNQIKI LGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDT YICEVEDQKEEVQLVVVGGGGGSGGGGSGGGGSDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:158 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSGGGGSGGGGSGGGGSDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:159 DNYWS SEQIDNO:160 YVHDSGDTNYNPSLKS SEQIDNO:161 TKHGRRIYGVVAFKEWFTYFYMDV SEQIDNO:162 GEESLGSRSVI SEQIDNO:163 NNNDRPS SEQIDNO:164 HIWDSRRPTNWV SEQIDNO:165 DAYWS SEQIDNO:166 YVHHSGDTNYNPSLKR SEQIDNO:167 ALHGKRIYGIVALGELFTYFYMDV SEQIDNO:168 GKESIGSRAVQ SEQIDNO:169 NNQDRPA SEQIDNO:170 HIYDARGGTNWV SEQIDNO:171 ACTYFWG SEQIDNO:172 SLSHCQSFWGSGWTFHNPSLKS SEQIDNO:173 FDGEVLVYNHWPKPAWVDL SEQIDNO:174 NGTATNFVS SEQIDNO:175 GVDKRPP SEQIDNO:176 GSLVGNWDVI SEQIDNO:177 ACDYFWG SEQIDNO:178 GLSHCAGYYNTGWTYHNPSLKS SEQIDNO:179 FDGEVLVYHDWPKPAWVDL SEQIDNO:180 TGTSNRFVS SEQIDNO:181 GVNKRPS SEQIDNO:182 SSLVGNWDVI SEQIDNO:183 RCNYFWG SEQIDNO:184 SLSHCRSYYNTDWTYHNPSLKS SEQIDNO:185 FGGEVLVYRDWPKPAWVDL SEQIDNO:186 TGTSNNFVS SEQIDNO:187 EVNKRPS SEQIDNO:188 SSLVGNWDVI SEQIDNO:189 TGHYYWG SEQIDNO:190 HIHYTTAVLHNPSLKS SEQIDNO:191 SGGDILYYYEWQKPHWFSP SEQIDNO:192 NGTSSDIGGWNFVS SEQIDNO:193 EVNKRPS SEQIDNO:194 SSLFGRWDVV SEQIDNO:185 TGHHYWG SEQIDNO:196 HIHYNTAVLHNPALKS SEQIDNO:197 SGGDILYYIEWQKPHWFYP SEQIDNO:198 SGTGSDIGSWNFVS SEQIDNO:199 EVNRRRS SEQIDNO:200 SSLSGRWDIV SEQIDNO:201 GTDWGENDFHYG SEQIDNO:202 SIHWRGRTTHYKTSFRS SEQIDNO:203 HKYHDIFRVVPVAGWFDP SEQIDNO:204 RASQNVKNNLA SEQIDNO:205 DASSRAG SEQIDNO:206 QQYEEWPRT SEQIDNO:207 GGEWGDSDYHWG SEQIDNO:208 SIHWRGTTHYNAPFRG SEQIDNO:209 HKYHDIVMVVPIAGWFDP SEQIDNO:210 RASQSVKNNLA SEQIDNO:211 DTSSRAS SEQIDNO:212 QQYEEWPRT SEQIDNO:213 DVWLN SEQIDNO:214 RIKSRTDGGTTDYAASVKG SEQIDNO:215 DGFIMIRGVSEDYYYYYMDV SEQIDNO:216 SGSSSNIGNNYVL SEQIDNO:217 GNNKRPS SEQIDNO:218 ATWDSGLSADWV SEQIDNO:219 SYVMH SEQIDNO:220 AISSDGETTYHANSVKG SEQIDNO:221 DRYYETSGSNAFDV SEQIDNO:222 QASQDISNYLN SEQIDNO:223 TASNLET SEQIDNO:224 QQYDNLGDLS SEQIDNO:225 NFAIH SEQIDNO:226 GRVPVVGIYKYGKKFHD SEQIDNO:227 WRGCGMCPYDTSSYYNDASDV SEQIDNO:228 RASQNISSSWIA SEQIDNO:229 AASARAA SEQIDNO:230 QYYGGSFFT SEQIDNO:231 AHTMN SEQIDNO:232 SISTSSTYRDYADAVKG SEQIDNO:233 KGSDRLSDNDPFDA SEQIDNO:234 RASQSIETWLA SEQIDNO:235 KASTLKT SEQIDNO:236 QHYAGYSAT SEQIDNO:237 SSYWS SEQIDNO:238 YTHHSGDTNYAPSLKS SEQIDNO:239 TLHGRRIYGVVAFNEFFTYFYWEV SEQIDNO:240 GGESIGSRAVQ SEQIDNO:241 NNQDRPP SEQIDNO:242 HIWDSRRPTNWV SEQIDNO:243 SSYWS SEQIDNO:244 YTHHSGDTNYAPSLKS SEQIDNO:245 TLHGRRIYGVVAFNEYYTYFYWPT SEQIDNO:246 GGESIGSRAVQ SEQIDNO:247 NNQDRPP SEQIDNO:248 HIWDSRRPTNWE SEQIDNO:249 SSYWS SEQIDNO:250 YTHHSGDTNYAPSLKS SEQIDNO:251 TLHGRRIYGVVAFNEYYTYFYWPT SEQIDNO:252 TGTSSDIGASDYVS SEQIDNO:253 DVTKRPS SEQIDNO:254 SSDAGRHTLL SEQIDNO:255 QVHLQESGPGLVKPSETLSLTCNVSGTLVRDNYWSWIRQPLGKQP EWIGYVHDSGDTNYNPSLKSRVHLSLDKSKNLVSLRLTGVTAADS AIYYCATTKHGRRIYGVVAFKEWFTYFYMDVWGKGTSVTVSS SEQIDNO:256 TFVSVAPGQTARITCGEESLGSRSVIWYQQRPGQAPSLIIYNNND RPSGIPDRFSGSPGSTFGTTATLTITSVEAGDEADYYCHIWDSRR PTNWVFGEGTTLIVL SEQIDNO:257 QVHLQESGPGLVKPSETLSLTCNVSGTLVRDNYWSWIRQPLGKQP EWIGYVHDSGDTNYNPSLKSRVHLSLDKSKNLVSLRLTGVTAADS AIYYCATTKHGRRIYGVVAFKEWFTYFYMDVWGKGTSVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:258 QVHLQESGPGLVKPSETLSLTCNVSGTLVRDNYWSWIRQPLGKQP EWIGYVHDSGDTNYNPSLKSRVHLSLDKSKNLVSLRLTGVTAADS AIYYCATTKHGRRIYGVVAFKEWFTYFYMDVWGKGTSVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:259 TFVSVAPGQTARITCGEESLGSRSVIWYQQRPGQAPSLIIYNNND RPSGIPDRFSGSPGSTFGTTATLTITSVEAGDEADYYCHIWDSRR PTNWVFGEGTTLIVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:260 QLHLQESGPGLVKPPETLSLTCSVSGASINDAYWSWIRQSPGKRP EWVGYVHHSGDTNYNPSLKRRVTFSLDTAKNEVSLKLVDLTAADS ATYFCARALHGKRIYGIVALGELFTYFYMDVWGKGTAVTVSS SEQIDNO:261 SSMSVSPGETAKISCGKESIGSRAVQWYQQKPGQPPSLIIYNNQD RPAGVPERFSASPDFRPGTTATLTITNVDAEDEADYYCHIYDARG GTNWVFDRGTTLTVL SEQIDNO:262 QLHLQESGPGLVKPPETLSLTCSVSGASINDAYWSWIRQSPGKRP EWVGYVHHSGDTNYNPSLKRRVTFSLDTAKNEVSLKLVDLTAADS ATYFCARALHGKRIYGIVALGELFTYFYMDVWGKGTAVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:263 QLHLQESGPGLVKPPETLSLTCSVSGASINDAYWSWIRQSPGKRP EWVGYVHHSGDTNYNPSLKRRVTFSLDTAKNEVSLKLVDLTAADS ATYFCARALHGKRIYGIVALGELFTYFYMDVWGKGTAVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:264 SSMSVSPGETAKISCGKESIGSRAVQWYQQKPGQPPSLIIYNNQD RPAGVPERFSASPDFRPGTTATLTITNVDAEDEADYYCHIYDARG GTNWVFDRGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:265 QSQLQESGPRLVEASETLSLTCNVSGESTGACTYFWGWVRQAPGK GLEWIGSLSHCQSFWGSGWTFHNPSLKSRLTISLDTPKNQVFLKL TSLTAADTATYYCARFDGEVLVYNHWPKPAWVDLWGRGIPVTVTV SS SEQIDNO:266 QSALTQPPSASGSPGQSITISCNGTATNFVSWYQQFPDKAPKLII FGVDKRPPGVPDRFSGSRSGTTASLTVSRLQTDDEAVYYCGSLVG NWDVIFGGGTTLTVL SEQIDNO:267 QSQLQESGPRLVEASETLSLTCNVSGESTGACTYFWGWVRQAPGK GLEWIGSLSHCQSFWGSGWTFHNPSLKSRLTISLDTPKNQVFLKL TSLTAADTATYYCARFDGEVLVYNHWPKPAWVDLWGRGIPVTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK SEQIDNO:268 QSQLQESGPRLVEASETLSLTCNVSGESTGACTYFWGWVRQAPGK GLEWIGSLSHCQSFWGSGWTFHNPSLKSRLTISLDTPKNQVFLKL TSLTAADTATYYCARFDGEVLVYNHWPKPAWVDLWGRGIPVTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGK SEQIDNO:269 QSALTQPPSASGSPGQSITISCNGTATNFVSWYQQFPDKAPKLII FGVDKRPPGVPDRFSGSRSGTTASLTVSRLQTDDEAVYYCGSLVG NWDVIFGGGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:270 QPQLQESGPGLVEASETLSLTCTVSGDSTAACDYFWGWVRQPPGK GLEWIGGLSHCAGYYNTGWTYHNPSLKSRLTISLDTPKNQVFLKL NSVTAADTAIYYCARFDGEVLVYHDWPKPAWVDLWGRGTLVTVTV SS SEQIDNO:271 QSALTQPPSASGSPGQSISISCTGTSNRFVSWYQQHPGKAPKLVI YGVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCSSLVG NWDVIFGGGTKLTVL SEQIDNO:272 QPQLQESGPGLVEASETLSLTCTVSGDSTAACDYFWGWVRQPPGK GLEWIGGLSHCAGYYNTGWTYHNPSLKSRLTISLDTPKNQVFLKL NSVTAADTAIYYCARFDGEVLVYHDWPKPAWVDLWGRGTLVTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK SEQIDNO:273 QPQLQESGPGLVEASETLSLTCTVSGDSTAACDYFWGWVRQPPGK GLEWIGGLSHCAGYYNTGWTYHNPSLKSRLTISLDTPKNQVFLKL NSVTAADTAIYYCARFDGEVLVYHDWPKPAWVDLWGRGTLVTVTV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGK SEQIDNO:274 QSALTQPPSASGSPGQSISISCTGTSNRFVSWYQQHPGKAPKLVI YGVNKRPSGVPDRFSGSKSGNTASLTVSGLQTDDEAVYYCSSLVG NWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:275 QPQLQESGPGLVEASETLSLTCTVSGDSTGRCNYFWGWVRQPPGK GLEWIGSLSHCRSYYNTDWTYHNPSLKSRLTISLDTPKNQVFLRL TSVTAADTATYYCARFGGEVLVYRDWPKPAWVDLWGRGTLVTVSS SEQIDNO:276 QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQYPGKAPKLVI YEVNKRPSGVPDRFSGSKSGSTASLTVSGLQADDEGVYYCSSLVG NWDVIFGGGTKLTVL SEQIDNO:277 QPQLQESGPGLVEASETLSLTCTVSGDSTGRCNYFWGWVRQPPGK GLEWIGSLSHCRSYYNTDWTYHNPSLKSRLTISLDTPKNQVFLRL TSVTAADTATYYCARFGGEVLVYRDWPKPAWVDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK SEQIDNO:278 QPQLQESGPGLVEASETLSLTCTVSGDSTGRCNYFWGWVRQPPGK GLEWIGSLSHCRSYYNTDWTYHNPSLKSRLTISLDTPKNQVFLRL TSVTAADTATYYCARFGGEVLVYRDWPKPAWVDLWGRGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK SEQIDNO:279 QSALTQPPSASGSPGQSITISCTGTSNNFVSWYQQYPGKAPKLVI YEVNKRPSGVPDRFSGSKSGSTASLTVSGLQADDEGVYYCSSLVG NWDVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:280 QVQLQESGPGLVKPAETLSLTCSVSGESINTGHYYWGWVRQVPGK GLEWIGHIHYTTAVLHNPSLKSRLTIKIYTLRNQITLRLSNVTAA DTAVYHCVRSGGDILYYYEWQKPHWFSPWGPGIHVTVSS SEQIDNO:281 QSALTQPPSASGSLGQSVTISCNGTSSDIGGWNFVSWYQQFPGRA PRLIIFEVNKRPSGVPGRFSGSKSGNSASLTVSGLQSDDEGQYFC SSLFGRWDVVFGGGTKLTVL SEQIDNO:282 QVQLQESGPGLVKPAETLSLTCSVSGESINTGHYYWGWVRQVPGK GLEWIGHIHYTTAVLHNPSLKSRLTIKIYTLRNQITLRLSNVTAA DTAVYHCVRSGGDILYYYEWQKPHWFSPWGPGIHVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK SEQIDNO:283 QVQLQESGPGLVKPAETLSLTCSVSGESINTGHYYWGWVRQVPGK GLEWIGHIHYTTAVLHNPSLKSRLTIKIYTLRNQITLRLSNVTAA DTAVYHCVRSGGDILYYYEWQKPHWFSPWGPGIHVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQ KSLSLSPGK SEQIDNO:284 QSALTQPPSASGSLGQSVTISCNGTSSDIGGWNFVSWYQQFPGRA PRLIIFEVNKRPSGVPGRFSGSKSGNSASLTVSGLQSDDEGQYFC SSLFGRWDVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKAT LVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:285 QVQLQESGPGLVKPSETLSLTCTVSGDSINTGHHYWGWVRQVPGK GPEWIAHIHYNTAVLHNPALKSRVTISIFTLKNLITLSLSNVTAA DTAVYFCVRSGGDILYYIEWQKPHWFYPWGPGILVTVSS SEQIDNO:286 QSALTQPPSASGSLGQSLTISCSGTGSDIGSWNFVSWYQQFPGRA PNLIIFEVNRRRSGVPDRFSGSKSGNTASLTVSGLRSEDEAEYFC SSLSGRWDIVFGGGTKVTVL SEQIDNO:287 QVQLQESGPGLVKPSETLSLTCTVSGDSINTGHHYWGWVRQVPGK GPEWIAHIHYNTAVLHNPALKSRVTISIFTLKNLITLSLSNVTAA DTAVYFCVRSGGDILYYIEWQKPHWFYPWGPGILVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK SEQIDNO:288 QVQLQESGPGLVKPSETLSLTCTVSGDSINTGHHYWGWVRQVPGK GPEWIAHIHYNTAVLHNPALKSRVTISIFTLKNLITLSLSNVTAA DTAVYFCVRSGGDILYYIEWQKPHWFYPWGPGILVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQ KSLSLSPGK SEQIDNO:289 QSALTQPPSASGSLGQSLTISCSGTGSDIGSWNFVSWYQQFPGRA PNLIIFEVNRRRSGVPDRFSGSKSGNTASLTVSGLRSEDEAEYFC SSLSGRWDIVFGGGTKVTVLGQPKAAPSVTLFPPSSEELQANKAT LVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:290 QLQLQESGPGLVKPSETLSLTCTVSGGSMRGTDWGENDFHYGWIR QSSAKGLEWIGSIHWRGRTTHYKTSFRSRATLSIDTSNNRFSLTF SFVTAADTAVYYCARHKYHDIFRVVPVAGWFDPWGQGLLVTVSS SEQIDNO:291 EIVMTQSPPTLSVSPGETATLSCRASQNVKNNLAWYQLKPGQAPR LLIFDASSRAGGIPDRFSGSGYGTDFTLTVNSVQSEDFGDYFCQQ YEEWPRTFGQGTKVDIK SEQIDNO:292 QLQLQESGPGLVKPSETLSLTCTVSGGSMRGTDWGENDFHYGWIR QSSAKGLEWIGSIHWRGRTTHYKTSFRSRATLSIDTSNNRFSLTF SFVTAADTAVYYCARHKYHDIFRVVPVAGWFDPWGQGLLVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPGK SEQIDNO:293 QLQLQESGPGLVKPSETLSLTCTVSGGSMRGTDWGENDFHYGWIR QSSAKGLEWIGSIHWRGRTTHYKTSFRSRATLSIDTSNNRFSLTF SFVTAADTAVYYCARHKYHDIFRVVPVAGWFDPWGQGLLVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALH SHYTQKSLSLSPGK SEQIDNO:294 EIVMTQSPPTLSVSPGETATLSCRASQNVKNNLAWYQLKPGQAPR LLIFDASSRAGGIPDRFSGSGYGTDFTLTVNSVQSEDFGDYFCQQ YEEWPRTFGQGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:295 EVHLEESGPGLVRPSETLSLTCTASGGSIRGGEWGDSDYHWGWVR HSPEKGLEWIGSIHWRGTTHYNAPFRGRGRLSIDLSRNQFSLRLT SVTAEDTAVYYCVKHKYHDIVMVVPIAGWFDPWGQGLQVTVSS SEQIDNO:296 EIMMTQSPAILSVSPGDRATLSCRASQSVKNNLAWYQKRPGQAPR LLIFDTSSRASGIPARFSGGGSGTEFTLTVNSMQSEDFATYYCQQ YEEWPRTFGQGTKVEIK SEQIDNO:297 EVHLEESGPGLVRPSETLSLTCTASGGSIRGGEWGDSDYHWGWVR HSPEKGLEWIGSIHWRGTTHYNAPFRGRGRLSIDLSRNQFSLRLT SVTAEDTAVYYCVKHKYHDIVMVVPIAGWFDPWGQGLQVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK SEQIDNO:298 EVHLEESGPGLVRPSETLSLTCTASGGSIRGGEWGDSDYHWGWVR HSPEKGLEWIGSIHWRGTTHYNAPFRGRGRLSIDLSRNQFSLRLT SVTAEDTAVYYCVKHKYHDIVMVVPIAGWFDPWGQGLQVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:299 EIMMTQSPAILSVSPGDRATLSCRASQSVKNNLAWYQKRPGQAPR LLIFDTSSRASGIPARFSGGGSGTEFTLTVNSMQSEDFATYYCQQ YEEWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:300 EVQLVESGGGLVKPGGSLRLTCVASGFTFSDVWLNWVRQAPGKGL EWVGRIKSRTDGGTTDYAASVKGRFTISRDDSKNTLYLQMNSLKT EDTAVYSCTTDGFIMIRGVSEDYYYYYMDVWGKGTTVTVSS SEQIDNO:301 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNNYVLWYQQFPGTAP KLLIYGNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYFCA TWDSGLSADWVFGGGTKLTVL SEQIDNO:302 EVQLVESGGGLVKPGGSLRLTCVASGFTFSDVWLNWVRQAPGKGL EWVGRIKSRTDGGTTDYAASVKGRFTISRDDSKNTLYLQMNSLKT EDTAVYSCTTDGFIMIRGVSEDYYYYYMDVWGKGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV DKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK SEQIDNO:303 EVQLVESGGGLVKPGGSLRLTCVASGFTFSDVWLNWVRQAPGKGL EWVGRIKSRTDGGTTDYAASVKGRFTISRDDSKNTLYLQMNSLKT EDTAVYSCTTDGFIMIRGVSEDYYYYYMDVWGKGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV DKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRT PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHY TQKSLSLSPGK SEQIDNO:304 QSVLTQPPSVSAAPGQKVTISCSGSSSNIGNNYVLWYQQFPGTAP KLLIYGNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYFCA TWDSGLSADWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKA TLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAAS SYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:305 QVQLVQSGGGLVQPGGSLRLSCAAFGFNFSSYVMHWVRQAPGQGL EYLSAISSDGETTYHANSVKGRFTSSRDNSKNTLFLQMGSLRTED VAVYYCARDRYYETSGSNAFDVWGQGTMVVVSS SEQIDNO:306 NSVLTQSPSSLSASVGDRVTITCQASQDISNYLNWYQHKPGKAPK LLIYTASNLETGVPSRFSGGGSGTHFSFTITSLQPEDAATYFCQQ YDNLGDLSFGGGTKVEIK SEQIDNO:307 QVQLVQSGGGLVQPGGSLRLSCAAFGFNFSSYVMHWVRQAPGQGL EYLSAISSDGETTYHANSVKGRFTSSRDNSKNTLFLQMGSLRTED VAVYYCARDRYYETSGSNAFDVWGQGTMVVVSSASTKGPSVFPLA PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK SEQIDNO:308 QVQLVQSGGGLVQPGGSLRLSCAAFGFNFSSYVMHWVRQAPGQGL EYLSAISSDGETTYHANSVKGRFTSSRDNSKNTLFLQMGSLRTED VAVYYCARDRYYETSGSNAFDVWGQGTMVVVSSASTKGPSVFPLA PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLS PGK SEQIDNO:309 NSVLTQSPSSLSASVGDRVTITCQASQDISNYLNWYQHKPGKAPK LLIYTASNLETGVPSRFSGGGSGTHFSFTITSLQPEDAATYFCQQ YDNLGDLSFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:310 QVQLVQSGAEVRKPGSSVTISCKPVGGTFTNFAIHWVRQAPGQGL EWVGGRVPVVGIYKYGKKFHDRLRLYEDDPMKTVFLELRSLTSDD TGVYYCTRWRGCGMCPYDTSSYYNDASDVWGPGTKVIVSA SEQIDNO:311 EIVLTQSPVTLSLSSGETGTLSCRASQNISSSWIAWYQQRRGQVP RLLISAASARAAGIPDRFTGRGSGTDFTLTITRLEPEDFGVYSCQ YYGGSFFTFGPGTQVDVK SEQIDNO:312 QVQLVQSGAEVRKPGSSVTISCKPVGGTFTNFAIHWVRQAPGQGL EWVGGRVPVVGIYKYGKKFHDRLRLYEDDPMKTVFLELRSLTSDD TGVYYCTRWRGCGMCPYDTSSYYNDASDVWGPGTKVIVSAASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK SEQIDNO:313 QVQLVQSGAEVRKPGSSVTISCKPVGGTFTNFAIHWVRQAPGQGL EWVGGRVPVVGIYKYGKKFHDRLRLYEDDPMKTVFLELRSLTSDD TGVYYCTRWRGCGMCPYDTSSYYNDASDVWGPGTKVIVSAASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYT QKSLSLSPGK SEQIDNO:314 EIVLTQSPVTLSLSSGETGTLSCRASQNISSSWIAWYQQRRGQVP RLLISAASARAAGIPDRFTGRGSGTDFTLTITRLEPEDFGVYSCQ YYGGSFFTFGPGTQVDVKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:315 EVQLVESGGGLVKAGGSLILSCGVSNFRISAHTMNWVRRVPGGGL EWVASISTSSTYRDYADAVKGRFTVSRDDLEDFVYLQMHKMRVED TAIYYCARKGSDRLSDNDPFDAWGPGTVVTVSP SEQIDNO:316 DVVMTQSPSTLSASVGDTITITCRASQSIETWLAWYQQKPGKAPK LLIYKASTLKTGVPSRFSGSGSGTEFTLTISGLQFDDFATYHCQH YAGYSATFGQGTRVEIK SEQIDNO:317 EVQLVESGGGLVKAGGSLILSCGVSNFRISAHTMNWVRRVPGGGL EWVASISTSSTYRDYADAVKGRFTVSRDDLEDFVYLQMHKMRVED TAIYYCARKGSDRLSDNDPFDAWGPGTVVTVSPASTKGPSVFPLA PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSWVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK SEQIDNO:318 EVQLVESGGGLVKAGGSLILSCGVSNFRISAHTMNWVRRVPGGGL EWVASISTSSTYRDYADAVKGRFTVSRDDLEDFVYLQMHKMRVED TAIYYCARKGSDRLSDNDPFDAWGPGTVVTVSPASTKGPSVFPLA PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSWVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLS PGK SEQIDNO:319 DVVMTQSPSTLSASVGDTITITCRASQSIETWLAWYQQKPGKAPK LLIYKASTLKTGVPSRFSGSGSGTEFTLTISGLQFDDFATYHCQH YAGYSATFGQGTRVEIKRTVAAPSVFIFPPSDEQLKSGTASWVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:320 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVHLGLHPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEFFTYFYWEVWGKGTQVTVSS SEQIDNO:321 SDISVAPGETVRISCGGESIGSRAVQWYQHRAGQAPKLIIYNNQD RPPGIPERFSGSPDIDFGTTATLTITNVEAGDEATYYCHIWDSRR PTNWVFGGGTTLTVL SEQIDNO:322 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVHLGLHPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEFFTYFYWEVWGKGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:323 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVHLGLHPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEFFTYFYWEVWGKGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:324 SDISVAPGETVRISCGGESIGSRAVQWYQHRAGQAPKLIIYNNQD RPPGIPERFSGSPDIDFGTTATLTITNVEAGDEATYYCHIWDSRR PTNWVFGGGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:325 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVHLGLHPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSS SEQIDNO:326 SDISVAPGETVRITCGGESIGSRAVQWYQHRPGQAPRLIIYNNQD RPPGIPERFSGSPDIDFGTTATLTISNVEAGDEATYYCHIWDSRR PTNWELGPGTTLTVL SEQIDNO:327 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVHLGLHPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:328 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVHLGLHPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:329 SDISVAPGETVRITCGGESIGSRAVQWYQHRPGQAPRLIIYNNQD RPPGIPERFSGSPDIDFGTTATLTISNVEAGDEATYYCHIWDSRR PTNWELGPGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:330 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVTIGLDPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSS SEQIDNO:331 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVTIGLDPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK SEQIDNO:332 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVTIGLDPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGK SEQIDNO:333 QMQLQESGPGLVKPGETLSLTCSVSGASISSSYWSWLRETPGKGL EWIGYTHHSGDTNYAPSLKSRVTIGLDPSKNQVSLSLTSVTAADT AVYYCARTLHGRRIYGVVAFNEYYTYFYWPTWGKGTQVTVSS SEQIDNO:334 QSVLTQPPSASGSPGQSVTISCTGTSSDIGASDYVSWYQQYPGEA PKVIIYDVTKRPSGVPDRFSGSKSGTTASLTVSGLQAEDEADYYC SSDAGRHTLLFGGGTKVTVL SEQIDNO:335 QVQLLESGPGLVRPSETLTLTCSVFNSRVSGYYYSWIRQPPGRGL EWIASTHFSLRPSRNPSLLSRVTTSIDTERYQVFLNMRSVTAADT AVYFCARGDASGWRADYFPHWGQGTLVVVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K SEQIDNO:336 QVQLLESGPGLVRPSETLTLTCSVFNSRVSGYYYSWIRQPPGRGL EWIASTHFSLRPSRNPSLLSRVTTSIDTERYQVFLNMRSVTAADT AVYFCARGDASGWRADYFPHWGQGTLVVVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPG K SEQIDNO:337 QSVLTQPPSASGSPGQSVTISCTGTSSDIGASDYVSWYQQYPGEA PKVIIYDVTKRPSGVPDRFSGSKSGTTASLTVSGLQAEDEADYYC SSDAGRHTLLFGGGTKVTVLGQPKAAPSVTLFPPSSEELQANKAT LVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQIDNO:338 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVHLQESGPGLVKPSETLSLTCNVSGTLVRD NYWSWIRQPLGKQPEWIGYVHDSGDTNYNPSLKSRVHLSLDKSKN LVSLRLTGVTAADSAIYYCATTKHGRRIYGVVAFKEWFTYFYMDV WGKGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:339 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQLHLQESGPGLVKPPETLSLTCSVSGASIND AYWSWIRQSPGKRPEWVGYVHHSGDTNYNPSLKRRVTFSLDTAKN EVSLKLVDLTAADSATYFCARALHGKRIYGIVALGELFTYFYMDV WGKGTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:340 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQSQLQESGPRLVEASETLSLTCNVSGESTGA CTYFWGWVRQAPGKGLEWIGSLSHCQSFWGSGWTFHNPSLKSRLT ISLDTPKNQVFLKLTSLTAADTATYYCARFDGEVLVYNHWPKPAW VDLWGRGIPVTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:341 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQPQLQESGPGLVEASETLSLTCTVSGDSTAA CDYFWGWVRQPPGKGLEWIGGLSHCAGYYNTGWTYHNPSLKSRLT ISLDTPKNQVFLKLNSVTAADTAIYYCARFDGEVLVYHDWPKPAW VDLWGRGTLVTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLV KDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:342 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQPQLQESGPGLVEASETLSLTCTVSGDSTGR CNYFWGWVRQPPGKGLEWIGSLSHCRSYYNTDWTYHNPSLKSRLT ISLDTPKNQVFLRLTSVTAADTATYYCARFGGEVLVYRDWPKPAW VDLWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:343 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLQESGPGLVKPAETLSLTCSVSGESINT GHYYWGWVRQVPGKGLEWIGHIHYTTAVLHNPSLKSRLTIKIYTL RNQITLRLSNVTAADTAVYHCVRSGGDILYYYEWQKPHWFSPWGP GIHVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVLHEALHSHYTQKSLSLSPGK SEQIDNO:344 QVQLQESGPGLVKPAETLSLTCSVSGESINTGHYYWGWVRQVPGK GLEWIGHIHYTTAVLHNPSLKSRLTIKIYTLRNQITLRLSNVTAA DTAVYHCVRSGGDILYYYEWQKPHWFSPWGPGIHVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQ KSLSLSPGKGGGGSKKVVYGKCGDTVELTCTASQKKNIQFHWKNS NQIKILGNQGSFLTKGPSKLNDRVDSRRSLWDQGNFPLIIKNLKP EDSDTYICEVEDQKEEVQLVVVC SEQIDNO:345 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQSALTQPPSASGSLGQSVTISCNGTSSDIGG WNFVSWYQQFPGRAPRLIIFEVNKRPSGVPGRFSGSKSGNSASLT VSGLQSDDEGQYFCSSLFGRWDVVFGGGTKLTVLGQPKAAPSVTL FPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETT TPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVA PTECS SEQIDNO:346 QSALTQPPSASGSLGQSVTISCNGTSSDIGGWNFVSWYQQFPGRA PRLIIFEVNKRPSGVPGRFSGSKSGNSASLTVSGLQSDDEGQYFC SSLFGRWDVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKAT LVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSGGGGSKKVV YGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLTKGPS KLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQKEEVQ LVVVC SEQIDNO:347 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLQESGPGLVKPSETLSLTCTVSGDSINT GHHYWGWVRQVPGKGPEWIAHIHYNTAVLHNPALKSRVTISIFTL KNLITLSLSNVTAADTAVYFCVRSGGDILYYIEWQKPHWFYPWGP GILVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVLHEALHSHYTQKSLSLSPGK SEQIDNO:348 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQLQLQESGPGLVKPSETLSLTCTVSGGSMRG TDWGENDFHYGWIRQSSAKGLEWIGSIHWRGRTTHYKTSFRSRAT LSIDTSNNRFSLTFSFVTAADTAVYYCARHKYHDIFRVVPVAGWF DPWGQGLLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG TQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:349 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSEVHLEESGPGLVRPSETLSLTCTASGGSIRG GEWGDSDYHWGWVRHSPEKGLEWIGSIHWRGTTHYNAPFRGRGRL SIDLSRNQFSLRLTSVTAEDTAVYYCVKHKYHDIVMVVPIAGWFD PWGQGLQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:350 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSEVQLVESGGGLVKPGGSLRLTCVASGFTFSD VWLNWVRQAPGKGLEWVGRIKSRTDGGTTDYAASVKGRFTISRDD SKNTLYLQMNSLKTEDTAVYSCTTDGFIMIRGVSEDYYYYYMDVW GKGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:351 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLVQSGGGLVQPGGSLRLSCAAFGFNFSS YVMHWVRQAPGQGLEYLSAISSDGETTYHANSVKGRFTSSRDNSK NTLFLQMGSLRTEDVAVYYCARDRYYETSGSNAFDVWGQGTMVVV SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGK SEQIDNO:352 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLVQSGAEVRKPGSSVTISCKPVGGTFTN FAIHWVRQAPGQGLEWVGGRVPVVGIYKYGKKFHDRLRLYEDDPM KTVFLELRSLTSDDTGVYYCTRWRGCGMCPYDTSSYYNDASDVWG PGTKVIVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEP VTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAK TKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:353 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSEVQLVESGGGLVKAGGSLILSCGVSNFRISA HTMNWVRRVPGGGLEWVASISTSSTYRDYADAVKGRFTVSRDDLE DFVYLQMHKMRVEDTAIYYCARKGSDRLSDNDPFDAWGPGTVVTV SPASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGK SEQIDNO:354 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQMQLQESGPGLVKPGETLSLTCSVSGASISS SYWSWLRETPGKGLEWIGYTHHSGDTNYAPSLKSRVHLGLHPSKN QVSLSLTSVTAADTAVYYCARTLHGRRIYGVVAFNEFFTYFYWEV WGKGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:355 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQMQLQESGPGLVKPGETLSLTCSVSGASISS SYWSWLRETPGKGLEWIGYTHHSGDTNYAPSLKSRVHLGLHPSKN QVSLSLTSVTAADTAVYYCARTLHGRRIYGVVAFNEYYTYFYWPT WGKGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:356 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQMQLQESGPGLVKPGETLSLTCSVSGASISS SYWSWLRETPGKGLEWIGYTHHSGDTNYAPSLKSRVTIGLDPSKN QVSLSLTSVTAADTAVYYCARTLHGRRIYGVVAFNEYYTYFYWPT WGKGTQVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP EPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:357 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLLESGPGLVRPSETLTLTCSVFNSRVSG YYYSWIRQPPGRGLEWIASTHFSLRPSRNPSLLSRVTTSIDTERY QVFLNMRSVTAADTAVYFCARGDASGWRADYFPHWGQGTLVVVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQ PREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEAL HSHYTQKSLSLSPGK SEQIDNO:358 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQSVLTQPPSASGSPGQSVTISCTGTSSDIGA SDYVSWYQQYPGEAPKVIIYDVTKRPSGVPDRFSGSKSGTTASLT VSGLQAEDEADYYCSSDAGRHTLLFGGGTKVTVLGQPKAAPSVTL FPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETT TPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVA PTECS SEQIDNO:359 QVQLLESGPGLVRPSETLTLTCSVFNSRVSGYYYSWIRQPPGRGL EWIASTHFSLRPSRNPSLLSRVTTSIDTERYQVFLNMRSVTAADT AVYFCARGDASGWRADYFPHWGQGTLVVVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSWVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRD ELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPG KGGGGSKKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGN QGSFLTKGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYIC EVEDQKEEVQLVVVC SEQIDNO:360 QSVLTQPPSASGSPGQSVTISCTGTSSDIGASDYVSWYQQYPGEA PKVIIYDVTKRPSGVPDRFSGSKSGTTASLTVSGLQAEDEADYYC SSDAGRHTLLFGGGTKVTVLGQPKAAPSVTLFPPSSEELQANKAT LVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASS YLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSGGGGSKKVV YGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLTKGPS KLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQKEEVQ LVVVC SEQIDNO:361 CTRPNNNTRKSIHIGPGRAFYTTGEIIGDIRQAHC SEQIDNO:362 KKVVYGKCGDTVELTCTASQKKNIQFHWKNSNQIKILGNQGSFLT KGPSKLNDRVDSRRSLWDQGNFPLIIKNLKPEDSDTYICEVEDQK EEVQLVVVCGGGGSQVQLVQSGAQMKNPGASVKVSCAPSGYTFTD FYIHWLRQAPGQGLQWMGWMNPQTGRTNTARNFQGRVTMTRDTSI GTAYMELRSLTSDDTAIYYCTTGGWISLYYDSSYYPNFDHWGQGT LLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK PKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VLHEALHSHYTQKSLSLSPGK SEQIDNO:363 MKVMGTKKNYQHLWRWGIMLLGMLMMSSAAEQLWVTVYYGVPVWR EANTTLFCASDAKAYDTEVHNVWATHACVPTDPNPQEVVMGNVTE DFNMWKNNMVEQMHEDIISLWDQSLKPCVKLTPLCVTLHCTNVTI SSTNGSTANVTMREEMKNCSFNTTTVIRDKIQKEYALFYKLDIVP IEGKNTNTSYRLINCNTSVITQACPKVSFEPIPIHYCAPAGFAIL KCNNKTFNGKGPCRNVSTVQCTHGIKPVVSTQLLLNGSLAEEDII IRSENFTNNGKNIIVQLKEPVKINCTRPGNNTRRSINIGPGRAFY ATGAIIGDIRKAHCNISTEQWNNTLTQIVDKLREQFGNKTIIFNQ SSGGDPEVVMHTFNCGGEFFYCNSTQLFNSTWFNNGTSTWNSTAD NITLPCRIKQVINMWQEVGKAMYAPPIRGQIDCSSNITGLILTRD GGSNSSQNETFRPGGGNMKDNWRSELYKYKVVKIEPLGIAPTRAK RRVVQREKRAVTLGAVFLGFLGAAGSTMGAASLTLTVQARLLLSG IVQQQSNLLRAIEAQQHMLQLTVWGIKQLQARVLAIERYLKDQQL LGIWGCSGKLICTTTVPWNTSWSNKSYDYIWNNMTWMQWEREIDN YTGFIYTLIEESQNQQEKNELELLELDKWASLWNWFNITNWLWYI KLFIMIIGGLVGLRIVCAVLSIVNRVRQGYSPLSFQTRLPNPRGP DRPEETEGEGGERDRDRSARLVNGFLAIIWDDLRSLCLFSYHRLR DLLLIVARVVEILGRRGWEILKYWWNLLKYWSQELKNSAVSLLNV TAIAVAEGTDRVIEIVQRAVRAILHIPTRIRQGFERALL SEQIDNO:364 AEQLWVIVYYGVPVWREANTTLFCASDAKAYDTEVHNVWATHACV PTDPNPQEVVMGNVTEDFNMWKNNMVEQMHEDIISLWDQSLKPCV KLTPLCVTLHCTNVTISSTNGSTANVTMREEMKNCSFNTTTVIRD KIQKEYALFYKLDIVPIEGKNTNTSYRLINCNTSVITQACPKVSF EPIPIHYCAPAGFAILKCNNKTFNGKGPCRNVSTVQCTHGIKPVV STQLLLNGSLAEEDIIIRSENFTNNGKNIIVQLKEPVKINCTRPG NNTRRSINIGPGRAFYATGAIIGDIRKAHCNISTEQWNNTLTQIV DKLREQFGNKTIIFNQSSGGDPEVVMHTFNCGGEFFYCNSTQLFN STWENNGTSTWNSTADNITLPCRIKQVINMWQEVGKAMYAPPIRG QIDCSSNITGLILTRDGGSNSSQNETFRPGGGNMKDNWRSELYKY KVVKIEPLGIAPTRAKRRVVQREKR SEQIDNO:365 SDISVAPGETVRITCGGESIGSRAVQWYQHRPGQAPRLIIYNNQD RPPGIPERFSGSPDIDFGTTATLTISNVEAGDEATYYCHIWDSRR PTNWELGPGTTLTVL SEQIDNO:366 SDISVAPGETVRITCGGESIGSRAVQWYQHRPGQAPRLIIYNNQD RPPGIPERFSGSPDIDFGTTATLTISNVEAGDEATYYCHIWDSRR PTNWELGPGTTLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLI SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLT PEQWKSHRSYSCQVTHEGSTVEKTVAPTECS