SUPPRESSION OF CANCER

Abstract

Polypeptides for use in suppressing cancer and cancer disorders and methods of treatment using such polypeptides.

Claims

1. A polypeptide comprising: (i) a non-cytotoxic protease capable of cleaving a SNARE protein that is expressed in a cancer cell; (ii) a targeting moiety capable of binding to a binding site located on the surface of the cancer cell wherein the binding site is capable of undergoing endocytosis to be incorporated into an endosome within the cancer cell; and (iii) a translocation domain capable of translocating the protease from within an endosome, across the endosomal membrane and into the cytosol of the cancer cell; wherein the cancer cell is not a neuroendocrine tumour cell and the polypeptide is not clostridial neurotoxin.

2. (canceled)

3. The polypeptide of claim 1, wherein the cancer cell is selected from the group consisting of: a lung cancer cell, a renal cancer cell, a brain cancer cell, a breast cancer cell, a pancreatic cancer cell, a colorectal cancer cell, an adrenal cancer cell, an oesophageal cancer cell, a lymphoma cancer cell, a leukaemia cancer cell, an acute leukaemia cancer cell, a bladder cancer cell, a bone cancer cell, a bowel cancer cell, a cervical cancer cell, a chronic lymphocytic leukaemia cell, a Hodgkin's lymphoma cell, a liver cancer cell, a melanoma skin cancer cell, an oropharyngeal cancer cell, a myeloma cell, a prostate cancer cell, a soft tissue sarcoma cell, a gastric cancer cell, a testicular cancer cell, a uterine cancer cell and, a Kaposi sarcoma cancer cell.

4. The polypeptide of claim 1, wherein the targeting moiety binds to a receptor selected from the group consisting of: an ErbB receptor a growth hormone-releasing hormone (GHRH) receptor; an insulin-like growth factor receptor (IGFR); a gastrin-releasing peptide (GRP) receptor; a bombesin peptide (BB) receptor; a growth hormone (GH) receptor; a vascular endothelial growth factor (VEGF) receptor; an acetylcholine (ACH) receptor; a somatostatin (SST) receptor; a cortistatin (CST) receptor; a chemokine (C-X-C motif) receptor; a neuropilin receptor; a gonadotropin-releasing hormone (GnRH) receptor; a VIP-glucagon-GRF-secretin superfamily receptor; a FLT receptor; a CHRN receptor; an EPH receptor; an EFN receptor; a DLK1 receptor, a DLL3 receptor, a FGF receptor; a JAG receptor; a LIF receptor; a NMB receptor; a NOTCH receptor; a PDGF receptor; a c-kit receptor; a TGF receptor; an endothelin receptor; a chemokine receptor; and an angiopoietin receptor.

5. The polypeptide of claim 1, wherein the targeting moiety is selected from the group consisting of: an ErbB peptide, an adrenomedullin (ADM) peptide, an AM peptide, an AMF peptide, an amphiregulin peptide, an APRIL peptide, an artemin peptide, a betacellulin peptide, a bombesin peptide, a CTR peptide, an endothelin peptide, an erythropoietin peptide, a FGF peptide, a FSH peptide, a gastrin peptide, a gastrin releasing peptide (GRP), a GDNF peptide, a ghrelin peptide, a GHRH peptide, a G-CSF peptide, a growth hormone peptide, a HB-EGF peptide, a hepatocyte growth factor (HGF) peptide, an IL peptide, a keratinocyte growth factor peptide, a leptin peptide, a LIF peptide, an alpha-melanotropin peptide, a MGSA/GRO peptide, a NRG peptide, an oxytocin peptide, an osteopontin (OPN) peptide, a neuregulin-1 peptide, a VIP peptide, a PACAP peptide, a PDGF peptide, a prolactin peptide, a SCF peptide, a somatostatin (SST) peptide, a cortistatin (CST) peptide, a TNF peptide, a TGF peptide, a VEGF peptide, a vasopressin peptide, an angiopoietin peptide, a B-CLL peptide, a BCGF-12KD peptide, a BAFF peptide, a galanin peptide, a GDNF peptide, a GnRH peptide, an IGF-II peptide, a LH peptide, a neurotrophin peptide, a substance P peptide, a TGF-alpha peptide, an IGF peptide, an angiopoietin peptide, a CXC peptide, and a CCL peptide.

6. The polypeptide of claim 1, wherein the non-cytotoxic protease comprises a clostridial neurotoxin L-chain or an IgA protease.

7. The polypeptide of claim 1, wherein the translocation domain comprises a clostridial neurotoxin translocation domain.

8-12. (canceled)

13. The polypeptide of claim 1, wherein the polypeptide comprises an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 7, 10, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44.

14. A nucleic acid encoding the polypeptide of claim 1.

15. A nucleic acid encoding the polypeptide of claim 1, wherein the nucleic acid comprises a nucleic acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 6 or 9.

16. A method of suppressing a cancer in a patient, the method comprising administering to the patient an effective amount of the polypeptide of claim 1.

17. The method of claim 16, wherein the cancer is selected from the group consisting of: lung cancer, renal cancer, brain cancer, breast cancer, pancreatic cancer, colorectal cancer, adrenal cancer, oesophageal cancer, lymphoma, leukaemia, acute leukaemia, bladder cancer, bone cancer, bowel cancer, cervical cancer, chronic lymphocytic leukaemia, Hodgkin's lymphoma, liver cancer, skin cancer, oropharyngeal cancer, myeloma, prostate cancer, gastric cancer, testicular cancer, uterine cancer, and Kaposi sarcoma.

18. The method of claim 16, wherein the method comprises suppressing secretion from a cancer cell selected from the group consisting of: a lung cancer cell, a renal cancer cell, a brain cancer cell, a breast cancer cell, a pancreatic cancer cell, a colorectal cancer cell, an adrenal cancer cell, an oesophageal cancer cell, a lymphoma cancer cell, a leukaemia cancer cell, an acute leukaemia cancer cell, a bladder cancer cell, a bone cancer cell, a bowel cancer cell, a cervical cancer cell, a chronic lymphocytic leukaemia cell, a Hodgkin's lymphoma cell, a liver cancer cell, a skin cancer cell, an oropharyngeal cancer cell, a myeloma cell, a prostate cancer cell, a soft tissue sarcoma cell, a gastric cancer cell, a testicular cancer cell, a uterine cancer cell, and or a Kaposi sarcoma cell.

19. The polypeptide of claim 4, wherein the targeting moiety is an antibody or antibody fragment.

20. A method of suppressing a cancer in a patient, the method comprising administering to the patient an effective amount of the nucleic acid of claim 14.

21. The polypeptide of claim 1, wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to any one of SEQ ID NOs: 7, 10, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44.

22. The polypeptide of claim 1, wherein the polypeptide comprises an amino acid sequence having at least 98% sequence identity to any one of SEQ ID NOs: 7, 10, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44.

23. A nucleic acid encoding the polypeptide of claim 1, wherein the nucleic acid comprises a nucleic acid sequence having at least 95% sequence identity to any one of SEQ ID NOs: 6 or 9.

24. A nucleic acid encoding the polypeptide of claim 1, wherein the said nucleic acid comprises a nucleic acid sequence having at least 98% sequence identity to any one of SEQ ID NOs: 6 or 9.

Description

[0242] FIG. 1Purification of LH.sub.N/D-CT-CST29 fusion protein

[0243] Using the methodology outlined in Example 3, a LH.sub.N/D-CT-CST29 fusion protein was purified from E. coli BL21 (DE3) cells. Briefly, the soluble products obtained following cell disruption were applied to a nickel-charged affinity capture column. Bound proteins were eluted with 200 mM imidazole, treated with enterokinase to activate the fusion protein and then re-applied to a second nickel-charged affinity capture column. Samples from the purification procedure were assessed by SDS-PAGE. Lane 1: First nickel chelating Sepharose column eluant, Lane 2: Second nickel chelating Sepharose column eluant under non-reducing conditions, Lane 3: Second nickel chelating Sepharose column eluant under reducing conditions, Lane 4: Molecular mass markers (kDa).

[0244] FIG. 2Purification of LH.sub.N/A-CT-SST14 fusion protein

[0245] Using the methodology outlined in Example 3, an LH.sub.N/A-CT-SST14 fusion protein was purified from E. coli BL21 (DE3) cells. Briefly, the soluble products obtained following cell disruption were applied to a nickel-charged affinity capture column. Bound proteins were eluted with 200 mM imidazole, treated with Factor Xa to activate the fusion protein and then re-applied to a second nickel-charged affinity capture column. Samples from the purification procedure were assessed by SDS-PAGE. Lane 1: First nickel chelating Sepharose column eluant, Lane 2: Molecular mass markers (kDa), Lanes 3-4: Second nickel chelating Sepharose column eluant under non-reducing conditions, Lanes 5-6: Second nickel chelating Sepharose column eluant under reducing conditions.

[0246] FIGS. 3-47Box Plots showing up-regulation of SNARE protein mRNA in cancer cells

[0247] Statistical analysis of the differences in SNARE expression between normal and primary cancer tissues was completed through use of Oncomine algorithms (Compendia Bioscience, Ann Arbor, Mich., USA) and gene microarray analysis tool. SNARE mRNA expression was compared to the median expression of all other genes in the respective study, for which a Normalised expression value was generated. Only studies with analysis results with P<0.05 are illustrated. In more detail, FIGS. 3-10 illustrate SNAP-25 expression profiles in different cancer versus non-cancer cells. FIGS. 11-16 illustrate syntaxin-1 expression profiles in different cancer versus non-cancer cells. FIGS. 17-20 illustrate syntaxin-2 expression profiles in different cancer versus non-cancer cells. FIGS. 21-24 illustrate syntaxin-3 expression profiles in different cancer versus non-cancer cells. FIGS. 25-32 illustrate VAMP-1 expression profiles in different cancer versus non-cancer cells. FIGS. 33-38 illustrate VAMP-2 expression profiles in different cancer versus non-cancer cells. FIGS. 39-45 illustrate VAMP-3 expression profiles in different cancer versus non-cancer cells. In all cases, the data illustrate up-regulation of SNARE proteins in cancer cells. FIG. 46 illustrates a statistically significant difference in syntaxin-1mRNA expression level between patients with recurrent invasive ductal breast cancer at 5 years post diagnosis and those patients without recurrence. FIG. 47 illustrates a statistically significant difference in syntaxin-1mRNA expression level in breast cancer patients in whom a metastatic event has occurred at 5 years post diagnosis versus those without metastases. Data is presented in box-plot form (see FIG. 3 for explanation) and/or as a histogram, where each bar represents the normalised gene expression level in each patient tissue sample.

[0248] FIG. 48Profiling cell surface receptor and SNARE protein expression in Renal Cell carcinoma lines by Western blot analysis.

[0249] Protein extracts from eight human renal cell carcinoma (RCC) lines grown in vitro were prepared in standard Laemmli sample buffer (lanes 3-10). Protein extracts prepared from primary cultures of rat spinal cord neurons and dorsal root ganglia were included as controls. Proteins were separated on 10% SDS-polyacrylamide gels and electrophoretically transferred to a nitrocellulose membrane. After blocking of the membrane, primary antisera were used to probe for each specific protein and detection was enabled by a peroxidise-conjugated anti-species IgG. The receptor proteins detected were ErbB receptor (epidermal growth factor receptor, EGF) and growth hormone releasing hormone receptor (GHRHR). The SNARE proteins tested were SNAP-25, syntaxin-2 and syntaxin-3. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a protein loading control.

[0250] FIG. 49Detection of SNAP-25 protein cleavage in a Renal Cell carcinoma line by Western blot analysis after 24 hr treatment with an EGF-LHA fusion.

[0251] FIG. 49 shows the effect of 24 hour treatment of 786-0 human RCC cells with a number of EGF-liganded fusion protein molecules. In more detail, EGF-LHA, at doses between 1 and 100 nM, generated cleaved SNAP-25 species whereas control molecules, specifically EGF-LHB (non-SNAP-25 targeting), a catalytically inactive form of EGF-LHA, EGF-LH-DE-A (referred to as EGF-0 from hereon in) and the free EGF ligand, did not. Indicated concentrations are in nM.

[0252] FIG. 50Inhibition of in vitro proliferation of a Renal Cell carcinoma line by an EGF-LHA fusion.

[0253] 786-0 cells seeded into a cell culture vessel were counted in a pre-define region at 24 and 48 hours after treatment with 25 nM of EGF-liganded fusion proteins.

[0254] FIG. 51Inhibition of FGF-2 secretion from a Renal Cell carcinoma line by an EGF-LHA fusion.

[0255] Culture media from 786-0 cells treated for 24 hours with EGF-liganded fusion proteins were analysed for Fibroblast Growth Factor-2 by standard methods. In more detail, EGF-LHA, at doses between 1 and 50 nM, demonstrated a dose-dependent decrease in FGF-2 levels present in the culture medium whereas control molecules, specifically the catalytically inactive form of EGF-LHA, EGF-0, and free EGF ligand, did not. Indicated concentrations are in nM.

[0256] FIG. 52Inhibition of in vitro proliferation of the A498 Renal Cell carcinoma line by an EGF-LHA fusion.

[0257] FIG. 52 shows the effect of an EGF-liganded fusion protein (EGFv3-LHA, 300 nM) on cellular proliferation of A498 cell line after 48 hours. Cells were stained using the tetrazolium salt WST as per standard protocols. The unliganded molecule LHA was included as a control.

[0258] FIG. 53Inhibition of in vitro proliferation of the ACHN Renal Cell carcinoma line by an EGF-LHA fusion.

[0259] FIG. 53 shows the effect of an EGF-liganded fusion protein (EGFv3-LHA, at 300 nM) on cellular proliferation of ACHN cell line after 48 hours. Cells were stained using the tetrazolium salt WST as per standard protocols. The unliganded molecule LHA was included as a control.

[0260] FIG. 54Inhibition of in vitro secretion of gastrin releasing peptide (GRP) from the small cell lung cancer cell line DMS-53 by an EGF-LHA fusion.

[0261] FIG. 54 shows the effect of an EGF-liganded fusion protein (EGFv3-LHA, at 150 nM) on GRP secretion from the DMS-53 cell line after treatment for 24 hr. Medium removed from the cells were analysed 48 hours after removal of fusion proteins. The catalytically inactive form of EGF-LHA, EGF-0, was included as a control, as was cycloheximide, a general protein synthesis inhibitor.

NOMENCLATURE

[0262] SST somatostatin
TGF(A) transforming growth factor (alpha)
GHRL ghrelin
LEP leptin
ET(A) endothelin-1
FLT vascular endothelial growth factor receptor
CHRN(D) acetylcholine receptor (subunit delta)
EPHA ephrin type-A receptor
EFNA ephrin-A
DLK1 delta-like protein 1
JAG jagged protein
NRG neuregulin
G-CSF granulocyte colony-stimulating factor
AMF autocrine motility factor
NMB neuromedin-B
CCK cholecystokinin
PDGF platelet-derived growth factor
ADM adrenomedullin
GDNF glial cell line-derived neurotrophic factor
TrkA high affinity nerve growth factor
FSH follicle-stimulating hormone
CXCR C-X-C chemokine receptor
CRLR calcitonin-receptor-like receptor
PDF prostate differentiation factor
MCP monocyte chemotactic protein
KGF keratinocyte growth factor
FLK1 vascular endothelial growth factor receptor 2
PDGFR platelet-derived growth factor receptor
NOTCH neurogenic locus notch homolog protein
DLL delta-like protein
GHS growth hormone secretagogue
c-MET hepatocyte growth factor
c-kit mast/stem cell growth factor
MGSA/GRO melanoma growth stimulatory activity/growth related gene
BCGF B-cell growth factor
GnRH gonadotropin-releasing hormone receptor
Ang-2 angiopoietin-2
FGF fibroblast growth factor
ErbB epidermal growth factor receptor family member
VIPR vasoactive intestinal polypeptide receptor
BRS bombesin receptor subtype
GRP gastrin releasing peptide
LIF leukaemia inhibitory factor
GHRH growth hormone-releasing hormone
IGF insulin-like growth factor
CRHR-2 corticotropin releasing factor receptor-2
BB bombesin
GH growth hormone
IL interleukin
VEGF vascular endothelial growth factor
ACH acetylcholine
CST cortistatin
VPAC vasoactive intestinal peptide receptor
GRPR gastrin releasing peptide receptor
CTR calcitonin binding receptor
EPO erythropoietin
HB-EGF heparin-binding EGF-like growth factor
HGF/SF hepatocyte growth factor/scatter factor
SDF-1 stromal cell-derived factor 1
CXCL12 chemokine (C-X-C motif) ligand 12 (SDF-1)
TNF tumour necrosis factor
PGF placental growth factor
Gran4 granulin-4
TIE2 angiopoietin receptor-2
LH luteinising hormone
CCL CC chemokine ligand
NT neurotrophin
NTAK neuregulin-2
BAFF B-cell activating factor
GM-CSF granulocyte-macrophage colony stimulating factor
NGF nerve growth factor
PACAP pituitary adenylate cyclase-activating peptide
OB leptin
NRP neuropilin receptor

Summary of Examples

[0263] Example 1 Preparation of a LHA backbone construct

[0264] Example 2 Construction of LHD-CT-CST29

[0265] Example 3 Expression and purification of a LHD-CT-CST29 fusion protein

[0266] Example 4 Construction of LHA-CP-EGF

[0267] Example 5 Expression and purification of a LHA-CP-EGF fusion protein

[0268] Example 6 Chemical conjugation of LH.sub.N/A to GnRH TM

[0269] Example 7 Method for treating colorectal cancer

[0270] Example 8 Method for treating breast cancer

[0271] Example 9 Method for treating prostate cancer

[0272] Example 10 Method for treating lung carcinoid tumours

[0273] Example 11 Method for treating bladder cancer

[0274] Example 12 Method for treating small cell lung cancer

[0275] Example 13 Method for treating prostate cancer

[0276] Example 14 Method for treating cervical cancer

[0277] Example 15 Method for treating leukaemia

[0278] Example 16 Method for treating small cell lung cancer

[0279] Example 17 Method for treating pancreatic cancer

[0280] Example 18 Method for treating metastatic bone cancer

[0281] Example 19 Method for treating metastatic small cell lung cancer in the brain

[0282] Example 20 Method for treating bowel cancer

[0283] Example 21 Method for treating chronic lymphocytic leukaemia

[0284] Example 22 Method for treating liver cancer

[0285] Example 23 Method for treating Hodgkin's lymphoma

[0286] Example 24 Method for treating renal cancer

[0287] Example 25 Method for treating skin cancer

[0288] Example 26 Method for treating oropharyngeal cancer

[0289] Example 27 Method for treating myeloma cancer

[0290] Example 28 Method for treating soft tissue sarcoma cancer

[0291] Example 29 Method for treating gastric cancer

[0292] Example 30 Method for treating testicular cancer

[0293] Example 31 Method for treating uterine cancer

[0294] Example 32 Method for treating Karposi sarcoma

[0295] Example 33 Method for treating primary brain cancer

[0296] Example 34 Method for treating rectal cancer

[0297] Example 35 Assessment of proliferation changes, inhibition of cellular secretion and concomitant SNARE cleavage after treatment of in vitro cultured renal cancer cell lines with a polypeptide of the present invention.

Summary of SEQ ID NOs

[0298] Where an initial Met amino acid residue or a corresponding initial codon is indicated in any of the following SEQ ID NOs, said residue/codon is optional. [0299] 1. DNA sequence of LH.sub.N/A [0300] 2. DNA sequence of LH.sub.N/B [0301] 3. DNA sequence of LH.sub.N/C [0302] 4. DNA sequence of LH.sub.N/D [0303] 5. DNA sequence of the CT-CST29 linker [0304] 6. DNA sequence of the LHD-CT-CST29 fusion [0305] 7. Protein sequence of the LHD-CT-CST29 fusion [0306] 8. DNA sequence of the CP-EGF linker [0307] 9. DNA sequence of the LHA-CP-EGF fusion [0308] 10. Protein sequence of the LHA-CP-EGF fusion [0309] 11. Protein sequence of LH.sub.N/A [0310] 12. Protein sequence of LH.sub.N/B [0311] 13. Protein sequence of LH.sub.N/C [0312] 14. Protein sequence of LH.sub.N/D [0313] 15.Synthesised GnRH peptide [0314] 16. Protein sequence of the LHB-CT-SST28 fusion [0315] 17. Protein sequence of the LHA-CP-SST28 fusion [0316] 18. Protein sequence of the LHD-CT-EGF fusion [0317] 19. Protein sequence of the LHD-CT-VIP fusion [0318] 20. Protein sequence of the LHC-CT-IGF1 fusion [0319] 21. Protein sequence of the LHD-CT-IGF1 fusion [0320] 22. Protein sequence of the LHC-CT-VIP fusion [0321] 23. Protein sequence of the LHC-CT-GnRH fusion [0322] 24. Protein sequence of the LHD-CT-GnRH fusion [0323] 25. Protein sequence of the LHD-CT-GRP fusion [0324] 26. Protein sequence of the LHB-CT-GRP fusion [0325] 27. Protein sequence of the LHC-CT-LIF fusion [0326] 28. Protein sequence of the LHB-CP-LIF fusion [0327] 29. Protein sequence of the LHC-CT-FGF1 fusion [0328] 30. Protein sequence of the LHA-CP-FGF1 fusion [0329] 31. Protein sequence of the LHA-CT-FGF9 fusion [0330] 32. Protein sequence of the LHC-CP-FGF9 fusion [0331] 33. Protein sequence of the IgA-H.sub.Ntet-CT-SST14 fusion [0332] 34. Protein sequence of the IgA-H.sub.Ntet-CP-SST14 fusion [0333] 35. Protein sequence of the LHA-CT-SST14 fusion [0334] 36. Protein sequence of the LHA-CT-EGFv3 fusion [0335] 37. Protein sequence of the LHE-CT-IL6 fusion [0336] 38. Protein sequence of the LHB-CT-IL8 fusion [0337] 39. Protein sequence of the LHF-CP-GRAN4 fusion [0338] 40. Protein sequence of the LHD-CP-TGFa fusion [0339] 41. Protein sequence of the LHD-CP-TGFb fusion [0340] 42. Protein sequence of the LHB-CT-TNFa fusion [0341] 43. Protein sequence of the LHD-CT-SDF1 fusion [0342] 44. Protein sequence of the LHC-CT-VEGF fusion

EXAMPLES

Example 1 Preparation of a LH.SUB.N./a Backbone Construct

[0343] The following procedure creates a clone for use as an expression backbone for multidomain protein expression. This example is based on preparation of a serotype A based clone (SEQ ID1), though the procedures and methods are equally applicable to all LH.sub.N serotypes such as serotype B (SEQ ID2), serotype C (SEQ ID3) and serotype D (SEQ ID4) and other protease or translocation domains by using the appropriate published sequence for synthesis.

Preparation of Cloning and Expression Vectors

[0344] pCR 4 (Invitrogen) is the chosen standard cloning vector chosen due to the lack of restriction sequences within the vector and adjacent sequencing primer sites for easy construct confirmation. The expression vector is based on the pET (Novagen) expression vector which has been modified to contain the multiple cloning site NdeI-BamHI-SalI-PstI-XbaI-HindIII for construct insertion, a fragment of the expression vector has been removed to create a non-mobilisable plasmid, a variety of different fusion tags have been inserted to increase purification options and an existing XbaI site in the vector backbone has been removed to simplify sub-cloning.

Preparation of LC/A

[0345] The DNA sequence is designed by back translation of the LC/A amino acid sequence (obtained from freely available database sources such as GenBank (accession number P10845) using one of a variety of reverse translation software tools (for example Backtranslation tool v2.0 (Entelechon)). BamHI/SalI recognition sequences are incorporated at the 5 and 3 ends respectively of the sequence maintaining the correct reading frame. The DNA sequence is screened (using software such as SeqBuilder, DNASTAR Inc.) for restriction enzyme cleavage sequences incorporated during the back translation. Any cleavage sequences that are found to be common to those required by the cloning system are removed by the Backtranslation tool from the proposed coding sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the % GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, Sep. 13, 2004). This optimised DNA sequence containing the LC/A open reading frame (ORF) is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Preparation of H.SUB.N./a Insert

[0346] The DNA sequence is designed by back translation of the H.sub.N/A amino acid sequence (obtained from freely available database sources such as GenBank (accession number P10845) using one of a variety of reverse translation software tools (for example Back translation tool v2.0 (Entelechon)). A PstI restriction sequence added to the N-terminus and XbaI-stop codon-HindIII to the C-terminus ensuring the correct reading frame in maintained. The DNA sequence is screened (using software such as SeqBuilder, DNASTAR Inc.) for restriction enzyme cleavage sequences incorporated during the back translation. Any sequences that are found to be common to those required by the cloning system are removed by the Backtranslation tool from the proposed coding sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the % GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, Sep. 13, 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Preparation of the Interdomain (LC-H.SUB.N .Linker)

[0347] The LC-H.sub.N linker can be designed from first principle, using the existing sequence information for the linker as the template. For example, the serotype A linker (in this case defined as the inter-domain polypeptide region that exists between the cysteines of the disulphide bridge between LC and H.sub.N) has the sequence VRGIIPFKTKSLDEGYNKALNDL (SEQ ID NO: 111). This sequence information is freely available from available database sources such as GenBank (accession number P10845). For generation of a specific protease cleavage site, the native recognition sequence for Factor Xa can be used in the modified sequence VDGIITSKTKSLIEGRNKALNLQ (SEQ ID NO: 112) or an enterokinase recognition sequence is inserted into the activation loop to generate the sequence VDGIITSKTKSDDDDKNKALNLQ SEQ ID NO: 113). Using one of a variety of reverse translation software tools (for example Backtranslation tool v2.0 (Entelechon), the DNA sequence encoding the linker region is determined. BamHI/SalI and PstI/XbaI/stop codon/HindIII restriction enzyme sequences are incorporated at either end, in the correct reading frames. The DNA sequence is screened (using software such as Seqbuilder, DNASTAR Inc.) for restriction enzyme cleavage sequences incorporated during the back translation. Any sequences that are found to be common to those required by the cloning system are removed by the Backtranslation tool from the proposed coding sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the % GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, Sep. 13, 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Assembly and Confirmation of the Backbone Clone

[0348] Due to the small size, the activation linker must be transferred using a two step process. The pCR-4 linker vector is cleaved with BamHI+SalI combination restriction enzymes and the cleaved linker vector then serves as the recipient for BamHI+SalI restriction enzyme cleaved LC DNA. Once the LC encoding DNA is inserted upstream of the linker DNA, the entire LC-linker DNA fragment can then be isolated and transferred to the pET expression vector MCS. The LC-linker is cut out from the pCR 4 cloning vector using BamHI/PstI restriction enzymes digests. The pET expression vector is digested with the same enzymes but is also treated with antarctic phosphatase as an extra precaution to prevent re-circularisation. The LC-linker and the pET vector backbone are gel purified and the purified insert and vector backbone are ligated together using T4 DNA ligase. The product is transformed with TOP10 cells which are then screened for LC-linker using BamHI/PstI restriction digestion. The process is then repeated for the H.sub.N insertion into the PstI/HindIII restriction sites of the pET-LC-linker construct.

[0349] Screening with restriction enzymes is sufficient to ensure the final backbone is correct as all components are already sequenced confirmed during synthesis. However, during the sub-cloning of some components into the backbone, where similar size fragments are being removed and inserted, sequencing of a small region to confirm correct insertion is required.

Example 2 Construction of LH.SUB.N./D-CT-CST29

[0350] The following procedure creates a clone for use as an expression construct for multidomain fusion expression where the targeting moiety (TM) is presented C-terminally to the translocation domain. This example is based on preparation of the LH.sub.N/D-CT-CST29 fusion (SEQ ID6), though the procedures and methods are equally applicable to create other protease, translocation and TM fusions, where the TM of C-terminal to the translocation domain. In this example, a flanking 15 amino acid glycine-serine spacer is engineered into the interdomain sequence to ensure accessibility of the ligand to its receptor, but other spacers are applicable.

Preparation of Spacer-CST29 Insert

[0351] For presentation of a CST29 sequence at the C-terminus of the H.sub.N domain, a DNA sequence is designed to flank the spacer and targeting moiety (TM) regions allowing incorporation into the backbone clone (SEQ ID4). The DNA sequence can be arranged as BamHI-SalI-PstI-XbaI-spacer-CST29-stop codon-HindIII (SEQ ID5). The DNA sequence can be designed using one of a variety of reverse translation software tools (for example EditSeq best E. coli reverse translation (DNASTAR Inc.), or Backtranslation tool v2.0 (Entelechon)). Once the TM DNA is designed, the additional DNA required to encode the preferred spacer is created in silico. It is preferred to ensure the correct reading frame is maintained for the spacer, TM and restriction sequences and that the XbaI sequence is not preceded by the bases TC, which would result in DAM methylation. The DNA sequence is screened for restriction sequences incorporated and any additional sequences are removed manually from the remaining sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the % GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, Sep. 13, 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Assembly and Confirmation of the Backbone Clone

[0352] In order to create a LH.sub.N/D-CT-CST29 construct (SEQ ID6) using the backbone construct (SEQ ID4) and the newly synthesised pCR 4-spacer-TM vector encoding the CST29 TM (SEQ ID5), a one or two step method can be used; typically the two step method is used when the TM DNA is less than 100 base pairs. Using the one step method the TM can be inserted directly into the backbone construct buy cutting the pCR 4-spacer-TM vector with XbaI and HindIII restriction enzymes and inserting the TM encoding DNA fragment into a similarly cut pET backbone construct. Using the two-step method the LH.sub.N domain is excised from the backbone clone using restriction enzymes BamHI and XbaI and ligated into similarly digested pCR 4-spacer-TM vector. This creates an LH.sub.N-spacer-TM ORF in pCR 4 that can be excised from the vector using restriction enzymes BamHI and HindIII for subsequent ligation into the similarly cleaved pET expression construct. The final construct contains the LC-linker-H.sub.N-spacer-CST29 DNA (SEQ ID6) which will result in a fusion protein containing the sequence illustrated in SEQ ID7.

Example 3 Expression and Purification of a LH.SUB.N./D-CT-CST29 Fusion Protein

[0353] This example is based on preparation of an LH.sub.N/D protein that incorporates a CST29 TM polypeptide at the carboxyl terminus of the H.sub.N domain (SEQ ID7), where the pET expression vector ORF also encodes a histidine purification tag. These procedures and methods are equally applicable to fusion protein sequences shown in SEQ ID 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 31, 33, 35, 36, 37, 38, 42, 43 or 44. Where appropriate, the activation enzyme should be selected to be compatible with the protease activation site within each sequence.

Expression of LH.SUB.N./D-CT-CST29

[0354] Expression of the LH.sub.N/D-CT-CST29 protein is achieved using the following protocol. Inoculate 100 ml of modified TB containing 0.2% glucosamine and 30 g/ml kanamycin in a 250 ml flask with a single colony from the LH.sub.N/D-CT-CST29 expression strain. Grow the culture at 37 C., 225 rpm for 16 hours. Inoculate 1 L of modified TB containing 0.2% glucosamine and 30 g/ml kanamycin in a 2 L flask with 10 ml of overnight culture. Grow cultures at 37 C. until an approximate OD.sub.600 nm of 0.5 is reached at which point reduce the temperature to 16 C. After 1 hour induce the cultures with 1 mM IPTG and grow at 16 C. for a further 16 hours.

Purification of LH.SUB.N./D-CT-CST29 Protein

[0355] Defrost falcon tube containing 35 ml 50 mM HEPES pH 7.2 200 mM NaCl and approximately 10 g of E. coli BL21 (DE3) cell paste. Homogenise the cell paste (20 psi) ensuring the sample remains cool. Spin the lysed cells at 18 000 rpm, 4 C. for 30 minutes. Load the supernatant onto a 0.1 M NiSO.sub.4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCl. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. The eluted fusion protein is dialysed against 5 L of 50 mM HEPES pH 7.2 200 mM NaCl at 4 C. overnight and the OD.sub.280 nm measured to establish the protein concentration. Add 3.2 l enterokinase (New England Biolabs) per mg fusion protein and incubate static overnight at 25 C. Load onto a 0.1 M NiSO.sub.4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCl. Wash column to baseline with 50 mM HEPES pH 7.2 200 mM NaCl. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. Dialyse the eluted fusion protein against 5 L of 50 mM HEPES pH 7.2 150 mM NaCl at 4 C. overnight and concentrate the fusion to about 2 mg/ml, aliquot sample and freeze at 20 C. Test purified protein using OD.sub.280, BCA and purity analysis. FIGS. 1 and 2 demonstrate purification of fusion proteins following this method as analysed by SDS-PAGE.

Example 4 Construction of LH.SUB.N./A-CP-EGF

[0356] The following procedure creates a clone for use as an expression construct for multidomain fusion expression where the targeting moiety (TM) is presented centrally between the protease and translocation domain. This example is based on preparation of the LH.sub.N/A-CP-EGF fusion (SEQ ID9), though the procedures and methods are equally applicable to create other protease, translocation and TM fusions, where the TM is N-terminal to the translocation domain. In this example, a flanking helical spacer is engineered into the interdomain sequence to ensure accessibility of the ligand to its receptor, but other spacers are applicable.

Preparation of Spacer-Human EGF Insert

[0357] The LC-H.sub.N inter-domain polypeptide linker region exists between the cysteines of the disulphide bridge between LC and H.sub.N. For insertion of a protease cleavage site, spacer and a targeting moiety (TM) region into the activation loop, one of a variety of reverse translation software tools (for example Backtranslation tool v2.0 (Entelechon) are used to determine the DNA sequence encoding the linker region. For central presentation of a TM sequence at the N-terminus of the H.sub.N domain, a DNA sequence is designed for the spacer and targeting moiety (TM) regions allowing incorporation into the backbone clone (SEQ ID1). The DNA sequence can be arranged as BamHI-SalI-spacer-protease activation site-EGF-spacer-PstI-XbaI-stop codon-HindIII (SEQ ID8). Once the TM DNA is designed, the additional DNA required to encode the preferred spacer is created in silico. It is preferred to ensure the correct reading frame is maintained for the spacer, TM and restriction sequences and that the XbaI sequence is not preceded by the bases TC, which would result in DAM methylation. The DNA sequence is screened for restriction sequence incorporated and any additional sites are removed manually from the remaining sequence ensuring common E. coli codon usage is maintained. E. coli codon usage is assessed by reference to software programs such as Graphical Codon Usage Analyser (GeneArt), and the % GC content and codon usage ratio assessed by reference to published codon usage tables (for example GenBank Release 143, Sep. 13, 2004). This optimised DNA sequence is then commercially synthesized (for example by Entelechon, GeneArt or Sigma-Genosys) and is provided in the pCR 4 vector.

Assembly and Confirmation of the Backbone Clone

[0358] In order to create a LC-spacer-activation site-EGF-spacer-H.sub.N construct (SEQ ID9) using the backbone construct (SEQ ID1) and the newly synthesised pCR 4-spacer-activation site-TM-spacer vector encoding the EGF TM (SEQ ID8), a one or two step method can be used; typically the two step method is used when the TM DNA is less than 100 base pairs. Using the one step method the TM linker region can be inserted directly into the backbone construct buy cutting the pCR 4-spacer-activation site-TM-spacer vector with SalI and PstI restriction enzymes and inserting the TM encoding DNA fragment into a similarly cut pET backbone construct. Using the two-step method the LC domain is excised from the backbone clone using restriction enzymes BamHI and SalI and ligated into similarly digested pCR 4-spacer-activation site-TM-spacer vector. This creates a LC-spacer-activation site-TM-spacer ORF in pCR 4 that can be excised from the vector using restriction enzymes BamHI and PstI for subsequent ligation into similarly pET expression construct. The final construct contains the LC-spacer-activation site-EGF-spacer-H.sub.N DNA (SEQ ID9) which will result in a fusion protein containing the sequence illustrated in SEQ ID10.

Example 5 Expression and Purification of a LH.SUB.N./A-CP-EGF Fusion Protein

[0359] This example is based on preparation of an LH.sub.N/A protein that incorporates a EGF TM polypeptide into the interdomain linker region (SEQ ID10), where the pET expression vector ORF also encodes a histidine purification tag. These procedures and methods are equally applicable to the other fusion protein shown in SEQ ID 17, 28, 30, 32, 34, 39, 40 or 41. Where appropriate, the activation enzyme should be selected to be compatible with the protease activation site within each sequence.

Expression of LH.SUB.N./A-CP-EGF

[0360] Expression of the LH.sub.N/A-CP-EGF protein is achieved using the following protocol. Inoculate 100 ml of modified TB containing 0.2% glucosamine and 30 g/ml kanamycin in a 250 ml flask with a single colony from the LHA-CP-EGF expression strain. Grow the culture at 37 C., 225 rpm for 16 hours. Inoculate 1 L of modified TB containing 0.2% glucosamine and 30 g/ml kanamycin in a 2 L flask with 10 ml of overnight culture. Grow cultures at 37 C. until an approximate OD.sub.600 nm of 0.5 is reached at which point reduce the temperature to 16 C. After 1 hour induce the cultures with 1 mM IPTG and grow at 16 C. for a further 16 hours.

Purification of LH.SUB.N./A-CP-EGF Protein

[0361] Defrost falcon tube containing 35 ml 50 mM HEPES pH 7.2 200 mM NaCl and approximately 10 g of E. coli BL21 (DE3) cell paste. Homogenise the cell paste (20 psi) ensuring the sample remains cool. Spin the lysed cells at 18 000 rpm, 4 C. for 30 minutes. Load the supernatant onto a 0.1 M NiSO4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCl. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. The eluted fusion protein is dialysed against 5 L of 50 mM HEPES pH 7.2 200 mM NaCl at 4 C. overnight and the OD.sub.280 nm measured to establish the protein concentration. Add 3.2 l enterokinase (New England Biolabs) per mg fusion protein and incubate static overnight at 25 C. Load onto a 0.1 M NiSO4 charged Chelating column (20-30 ml column is sufficient) equilibrated with 50 mM HEPES pH 7.2 200 mM NaCl. Wash column to baseline with 50 mM HEPES pH 7.2 200 mM NaCl. Using a step gradient of 10, 40 and 100 mM imidazole, wash away the non-specific bound protein and elute the fusion protein with 200 mM imidazole. Dialyse the eluted fusion protein against 5L of 50 mM HEPES pH 7.2 150 mM NaCl at 4 C. overnight and concentrate the fusion to about 2 mg/ml, aliquot sample and freeze at 20 C. Test purified protein using OD.sub.280, BCA and purity analysis.

Example 6 Chemical Conjugation of LH.SUB.N./A to GnRH TM

[0362] The following procedure creates a chemically conjugated molecule containing the LH.sub.N/A amino acid sequence (SEQ ID11), prepared from SEQ ID1 using the production method outlined in example 3, and a GnRH peptide which has been chemically synthesised (SEQ ID15). However, the procedures and methods are equally applicable for the conjugation of other peptides to other protease/translocation domain proteins such as those containing the amino acid sequences SEQ ID12, 13 and 14.

[0363] The LH.sub.N/A protein was buffer exchanged from 50 mM Hepes 150 mM salt into PBSE (100 mM 14.2 g NA2HPO4, 100 mM 5.85 g NaCl, 1 mM EDTANa.sub.2 pH 7.5 with 1M HCl) using the Bio-rad PD10 column. This was done by washing one column volume of PBSE through the PD10 column, the protein was then added to the column until no more drops exit the end of the PD10 column. 8 mls of PBSE was then added and 0.5 ml fractions are collected. The collected fractions are the measured using the A.sub.280 reading and fractions containing protein are pooled. A concentration of 1.55 mg/ml of LH.sub.N/A was obtained from the buffer exchange step and this was used to set up the following reactions:

TABLE-US-00018 LH.sub.N/A 1.55 mg/ml 20 mM SPDP or Sulfo-LC-SPDP A 200 l 0 B 200 l 4 fold increase 0.62 l C 200 l 8 fold increase 1.24 l

[0364] Sample were left to tumble at RT for 3 hours before being passed down another PD10 column to buffer exchange into PBSE and the protein containing fractions pooled. A final concentration of 25 mM DTT was then added to derivatised protein and then the samples left at room temperature for 10 minutes. A.sub.280 and A.sub.343 readings were then taken to work out the ratio of SPDP:LH.sub.N/A interaction and the reaction which resulted in a derivatisation ration of between 1 and 3 was used for the peptide conjugation. The SPDP reagent binds to the primary amines of the LH.sub.N/A via an N-hydroxysuccinimide (NHS) ester, leaving the sulphydryl-reactive portion to form a disulphide bond to the free SH group on the free cysteine on the synthesised peptide. In this case the peptide sequence is GnRH which has been synthesised with a cysteine contained within the peptide to allow conjugation whilst leaving the N-terminus and C-terminus of the GnRH to interact with its receptor (SEQ ID15). The SPDP-derivatised LH.sub.N/A was mixed with a 4-fold excess of the GnRH ligand and the reaction was then left at RT for 90 minutes whilst tumbling. The excess GnRH was then removed using either a PD10 column leaving LH.sub.N/A-GnRH conjugated molecule.

Example 7Method for Treating Colorectal Cancer

[0365] A 62 year old man presents with a stage II colorectal cancer. To reduce and/or to prevent metastasis he receives a direct injection of a polypeptide of the present invention (eg. botulinum type A neurotoxin protease and translocation domain and a VIP peptide). Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is optionally performed in combination with chemotherapy, and is repeated 2 months later and 4 weeks later no tumour is observable anymore with the usual detection tools (colonoscopy, CT scan, PET scan, etc.) and the level of carcinoembryonic antigen (CEA) returned to the normal.

Example 8Method for Treating Breast Cancer

[0366] A 61 year old woman presents with a stage II breast cancer. To treat and/or prevent metastasis she receives an IV injection of a polypeptide of the present invention (eg. a botulinum type C neurotoxin protease, a botulinum type C neurotoxin translocation domain and a GnRH peptide), optionally in combination with chemotherapy. Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 6 weeks later no tumour is observable anymore with the usual detection tools (MRI, ultrasound, breast-specific positron emission tomography, mammography, Scintigraphy, etc).

Example 9Method for Treating Prostate Cancer

[0367] A 77 year old man presents with a stage II prostate cancer. To treatment and/or to prevent metastasis he receives a intravenous injection of a polypeptide of the present invention (eg. a botulinum type C neurotoxin protease, a botulinum type C neurotoxin translocation domain and an IGF-1 peptide), optionally in combination with hormone therapy. Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 8 weeks later no tumour is observable anymore with the usual detection tools (X-ray, ProstaScint scan, MRI, transrectal ultrasonography, CT scan, etc.) and the levels of PSA came back to normal.

Example 10Method for Treating Lung Carcinoid Tumours

[0368] A 66 year old woman presents with lung carcinoid tumours. To treat and/or to prevent metastasis she receives an IV injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain and a bFGF-1 peptide), optionally in combination with chemotherapy. Within 4 weeks a significant decrease in the size of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 1 month later and 4 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, bronchoscopy, etc.) or using the usual blood tests recommended for this cancer.

Example 11Method for Treating Bladder Cancer

[0369] A 56 year old man presents with a stage II bladder cancer. To treat and/or to prevent metastasis he receives a direct injection of a polypeptide of the present invention (eg. an IgA protease, a tetanus neurotoxin translocation domain and an IGF-1 peptide), optionally in combination with chemotherapy. Within 2 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 4 weeks later no tumour is observable anymore with the usual detection tools (colonoscopy, CT scan, PET scan, etc.).

Example 12Method for Treating Small Cell Lung Cancer

[0370] A 79 year old man is diagnosed with a stage I small cell lung cancer. To treat and/or to prevent metastasis he receives a injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain and a neuregulin ERBB3 peptide), optionally in combination with chemotherapy. Within 3 weeks a significant decrease in size of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 5 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, MRI, PET scanning, Radionuclide imaging, bronchoscopy, etc.) or using the usual blood tests recommended for this cancer.

Example 13Method for Treating Prostate Cancer

[0371] A 72 year old man is diagnosed with a stage II prostate cancer. To treat and/or to prevent metastasis he receives a intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and an IGF-1 peptide), optionally in combination with androgen deprivative treatment. Within 10 days a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 2 months later and 5 weeks later no tumour is observable anymore with the usual detection tools (X-ray, ProstaScint scan, MRI, transrectal ultrasonography, CT scan, etc.) and the levels of PSA came back to normal.

Example 14Method for Treating Cervical Cancer

[0372] A 60 year old woman diagnosed with cervical cancer at a limited stage is treated with surgery. To improve the effects of the treatment and to prevent metastasis she receives an intravenous injection of a polypeptide of the present invention (eg. botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain, a GS20 linker, and a somatostatin-14 peptide). Within 6 weeks no re-appearance of the tumour is observed. The treatment is repeated 3 months later and 8 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, MRI, PET scanning, Radionuclide imaging, bronchoscopy, etc.) or using the usual blood tests recommended for this cancer.

Example 15Method for Treating Leukaemia

[0373] A 42 year old man is diagnosed with a stage II Hairy cell leukaemia cancer. To treat and/or to prevent metastasis he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain, a GS20 linker, and a FGF-1 peptide), optionally in combination with chemotherapy. Within 10 days significant reduction in tumour burden is observed without appearance of metastasis. The treatment is repeated 1 month later and 3 weeks later no tumour is observable anymore with the usual detection tools (MRI, complete blood count, etc).

Example 16Method for Treating Small Cell Lung Cancer

[0374] A 56 year old man is diagnosed with a small cell lung cancer at an extensive stage. To treat and/or to prevent metastasis elsewhere he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain and an EGF peptide), optionally in combination with chemotherapy and radiation therapy. Within 3 weeks a significant shrinkage of the tumour and a diminution in size of the metastasis is observed without appearance of new metastasis elsewhere. The treatment is repeated twice after 2 months and 5 months. The patients died 11 months later, 6 months later than expected with this type of treatment and this stage of the disease.

Example 17Method for Treating Pancreatic Cancer

[0375] A 48 year old woman is diagnosed with pancreatic cancer at an advanced stage. She receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain and an EGF peptide), optionally in combination with the chemotherapeutic agent, Gemcitabine. Within 3 weeks a significant reduction in primary tumour growth and a diminution in size of the metastases are observed without appearance of new metastasis elsewhere. The treatment is repeated twice after 2 months and 5 months. The patient died 12 months later, 6 months later than expected with this type of treatment and this stage of the disease.

Example 18Method for Treating Metastatic Bone Cancer

[0376] A 71 year old man is diagnosed with a stage IV prostate cancer. To treat metastatic growth in his bone he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain and a TGF-beta peptide), optionally in combination with external beam radiation plus hormone therapy. Within 4 weeks a significant shrinkage of the tumour at the metastatic sites is observed without appearance of metastasis elsewhere. The treatment is repeated 2 and 4 months later. After 6 months no detectable increase in tumour burden is observable anymore with the usual detection tools (X-ray, ProstaScint scan, MRI, CT scan, etc.).

Example 19Method for Treating Metastatic Small Cell Lung Cancer in the Brain

[0377] A 65 year old man diagnosed with a small cell lung cancer at an advanced stage also presents with multiple brain metastases. To treat, a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain, and a SDF-1 peptide) is delivered into his brain by convection enhanced delivery, optionally in combination with chemotherapy and whole brain radiation. Within 4 weeks significant shrinkage of the metastatic tumour sites is observed without appearance of metastasis elsewhere. The treatment is repeated at 2 months and 8 weeks later no metastatic tumour is observable anymore with the usual detection tools (X-rays, CT scan, MRI, PET scanning, Radionuclide imaging, etc.) or using the usual blood tests recommended for this cancer.

Example 20Method for Treating Bowel Cancer

[0378] A 76 year old man is diagnosed with a stage II small bowel cancer. To treatment and/or to prevent metastasis he receives a direct injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease and translocation domain and an EGF peptide), optionally in combination with chemotherapy. Within 4 weeks a significant shrinkage of the tumour is observed without appearance of metastasis elsewhere and surgery is then realized to remove the tumour.

Example 21Method for Treating Chronic Lymphocytic Leukaemia

[0379] A 54 year old man is diagnosed with relapsed chronic lymphocytic leukaemia. To treat, he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type B neurotoxin protease, a botulinum type B neurotoxin translocation domain and an LIF-1 peptide), optionally in combination with chemotherapy. Within 2 weeks a significant reduction in tumour cell count in the patient's blood is observed which remains stable for an extended period of time as determined by microscopic examination and flow cytometric analysis of the patient's blood

Example 22Method for Treating Liver Cancer

[0380] A 55 year old woman is diagnosed with a stage IV hepatic cancer. To treat, she receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain and a TGF-alpha peptide), optionally in combination with chemotherapy. Within 2 weeks a significant shrinkage of the tumour is observed on PET-CT imaging studies along with a reduction of alpha-fetoprotein in the blood, which was elevated prior to treatment.

Example 23Method for Treating Hodgkin's Lymphoma

[0381] A 24 year old man is diagnosed with stage IVA Hodgkin's lymphoma. To treat, he receives repeated intravenous injections of a polypeptide of the present invention (eg. a botulinum type B neurotoxin protease, a botulinum type B neurotoxin translocation domain and an VEGF peptide), optionally in combination with chemotherapy. Within 2 weeks a significant reduction in tumour volume is observed and tumour blood flow using the usual detection tools (MRI, CT scans) and leads to complete remission within 2 months.

Example 24Method for Treating Renal Cancer

[0382] A 54 year old man is diagnosed with advanced renal cell carcinoma. To treat, he receives an intravenous administration of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type C neurotoxin translocation domain and a VEGF peptide), optionally in combination with the anti-angiogenic therapeutic, Sunitinib, a small molecule tyrosine kinase inhibitor (TKI). Within 14 days a significant shrinkage of the tumour over and above that expected with TKI alone is observed using the usual detection tools (CT, MRI scans, blood tests).

Example 25Method for Treating Skin Cancer

[0383] A 31 year old woman is diagnosed with a facial melanoma. To treat, she receives direct administration of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and a VEGF peptide), optionally in combination with chemotherapy and immunotherapy. Within 21 days a significant shrinkage of the tumour over and above that expected with chemo- and immunotherapy alone is observed, allowing surgical resection with clear but much reduced margins.

Example 26Method for Treating Oropharyngeal Cancer

[0384] A 63 year old man is diagnosed with a stage III head and neck cancer. To treat, he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type D neurotoxin translocation domain and a TGF-alpha peptide), optionally in combination with radiotherapy and chemotherapy. Within 4 weeks a significant improvement in the patient's ability to swallow food is apparent which is maintained longer than would be expected with the combination therapy alone.

Example 27Method for Treating Myeloma Cancer

[0385] A 73 year old woman diagnosed with multiple myeloma associated osteolytic bone lesions and hypercalcemia is treated with the standard chemotherapy. To improve the effects of the treatments she receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain and an IL-6 peptide). Within 4 weeks a significant reduction in the size of bone lesions and severity of hypercalcemia is observed without appearance of new lesions elsewhere using the usual detection tools (MRI, X-ray, blood tests).

Example 28Method for Treating Soft Tissue Sarcoma Cancer

[0386] A 51 year old woman diagnosed with a fibrosarcoma of the leg is treated with radiation therapy in an attempt to reduce tumour size prior to surgical resection. To improve the effects of the radiation treatment she receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and a bFGF peptide). Within 14 days a significant shrinkage of the tumour over and above that expected with radiation therapy alone is observed, allowing surgical resection with clear margins.

Example 29Method for Treating Gastric Cancer

[0387] A 84 year old man diagnosed with advanced gastric cancer and unable to undergo surgical resection is treated with radiation therapy to relieve tumour associated blockage. To improve the effects of the treatment he receives multiple direct injections of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain, a GS20 linker, and a GRP peptide). Within 5 days a significant shrinkage of the tumour is observed with the usual detection tools (gastroscopic examination and CT scan). The treatment is repeated 1 month later and 4 months later tumour blockage has not recurred.

Example 30Method for Treating Testicular Cancer

[0388] A 32 year old man is diagnosed with a stage I seminoma cancer. To treat and/or to prevent recurrence he receives a intravenous injection of a polypeptide of the present invention (eg. a botulinum type D neurotoxin protease, a botulinum type C neurotoxin translocation domain, a GS20 linker, and a VEGF peptide), optionally in combination with chemotherapy. Within 14 days a significant shrinkage of the tumour is observed. The treatment is repeated 1 month later and 6 weeks later no tumour is observable anymore with the usual detection tools (blood tests and CT scans).

Example 31Method for Treating Uterine Cancer

[0389] A 76 year old woman diagnosed with a stage IIA endometrial cancer is treated with usual chemotherapy and radiotherapy. To improve the effects of the treatment and to prevent metastasis she receives a direct injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease and translocation domain and an EGF peptide). Within 6 weeks a significant shrinkage of the tumour is observed on direct visualization of the uterine cavity by hysteroscopy without appearance of metastasis elsewhere.

Example 32Method for Treating Karposi Sarcoma

[0390] A 48 year old man is diagnosed with acquired immunity deficiency syndrome (AIDS) and presents with multiple Karposi sarcoma lesions. To treat, he receives an intravenous injection of a polypeptide of the present invention (eg. a botulinum type A neurotoxin protease, a botulinum type A neurotoxin translocation domain and an IL-6 peptide), optionally in combination with interferon alpha. Within 3 weeks a significant reduction in lesion size is observed without appearance of new lesions elsewhere. The treatment is repeated twice after 1 month and 3 months which effectively stops the progression of the Kaposi sarcoma.

Example 33Method for Treating Primary Brain Cancer

[0391] A 45 year old woman is diagnosed with glioblastoma. To treat and/or to prevent further metastasis she receives an intracranial application of a polypeptide of the present invention (eg. a botulinum type C neurotoxin protease, a botulinum type C neurotoxin translocation domain and a VEGF peptide), optionally in combination with chemotherapy. Within 4 weeks a significant decrease in the size of the tumour is observed without appearance of metastasis elsewhere. The treatment is repeated 1 month later and 4 weeks later no tumour is observable anymore with the usual detection tools (X-rays, CT scan, etc.).

Example 34Method for Treating Rectal Cancer

[0392] A 77 year old man diagnosed with a stage II rectal cancer is treated with a polypeptide of the present invention (eg. botulinum type A neurotoxin protease and an anti-EGFR antibody F(ab).sub.2 fragment), optionally incombination with radiotherapy and surgery. He receives localised injections of polypeptide (eg. 3 days in advance of a standard regiment of fractionated radiation) and within 2 weeks a significant shrinkage of the tumour is observed which enables complete surgical resection. 12 months later no tumour recurrence is observable with the usual detection tools (colonoscopy, CT scan, PET scan, etc.) and the level of carcinoembryonic antigen (CEA) returned to the normal.

Example 35Assessment of Proliferation Changes, Inhibition of Cellular Secretion and Concomitant SNARE Cleavage after Treatment of In Vitro Cultured Renal Cancer Cell Lines with a Polypeptide of the Present Invention

Methods:

Proliferation Analysis:

[0393] An appropriate volume containing 1000 cells of a suitable renal cancer cell line, for example A498, ACHN or 786-0, are seeded into the wells of a 96-well cell culture plate in a suitable growth medium supplemented with 10% Foetal Bovine Serum and incubated at 37 C. in a humidified atmosphere with 5% CO.sub.2. Cells are allowed to adhere overnight after which treatment with an EGF-liganded LHA molecule, such as an LHA molecule with a C-terminal presented EGF ligand, is initiated. After 24 hours, the treatment media are removed, cell monolayers are washed to remove traces of LHA-EGF and fresh medium applied to cells. After a further 24 hours a conventional calorimetric proliferation assay (based on the determination of the cleavage of the tetrazolium salt WST-1 to formazan by cellular enzymes) is performed. Specifically, WST-1 is added to the culture medium for 4 hours after which the optical density at 440 nm is determined for each treatment. FIGS. 50-52 demonstrate inhibition of in vitro proliferation from a renal cell carcinoma line by an EGF-LHA fusion.

Cellular Secretion Analysis:

[0394] 10,000 cells of a suitable renal cancer cell line (for example 786-0, A498 or ACHN) are seeded in the wells of a 24 well plate in an appropriate culture medium containing 10% foetal bovine serum. Plates are incubated overnight in an incubator at 37 C. in a humidified atmosphere with 5% CO.sub.2 to allow cells to adhere. Cell cultures are then treated with an appropriate polypeptide of the present invention which targets a specific receptor on the cells of interest (for the cell lines detailed above an appropriate molecule would be an EGF-liganded LHA as these 3 cell lines all express EGF receptors, FIG. 48). After 24 hours, the treatment medium is removed, cell monolayers refed with fresh culture medium and 24-48 hours later samples of the culture medium are removed to fresh microfuge tubes which are subsequently spun at 1500 rpm for 5 minutes to remove floating cells. The resultant supernatants are removed to fresh tubes and aliquots then used for the quantification of specific analytes (for example VEGF, TNF-alpha) by enzyme-linked immunosorbent assay or ELISA (FIG. 53). An example of analysis of inhibition of a secretion (GRP) from a small cell lung carcinoma cell line is provided in FIG. 54.

Western Blot Analysis to Detect SNARE Cleavage Due to Cellular Uptake

[0395] of Polypeptides of the Present Invention: Cell cultures are treated with polypeptides as detailed above for the analysis of cellular secretions. After removal of culture medium for subsequent ELISA analysis, cell monolayers in each well are washed three times with phosphate-buffered saline and protein extracts prepared by cellular lysis in standard Laemmli sample buffer. Cellular protein are separated on 12% SDS-polyacrylamide gels and electrophoretically transferred to a nitrocellulose membrane. After blocking of the membrane, primary antisera are used to probe for the SNARE protein of interest and detection of full length and/or cleaved forms is enabled by a peroxidase-conjugated anti-species IgG. In the case of treatment of the renal cell line 786-0 with an LHA-EGF molecule, cleavage of the SNARE SNAP-25 is observed (see FIG. 49).

TABLE-US-00019 SEQIDNOs 1.DNAsequenceofLH.sub.N/A ggatccATGGAGTTCGTTAACAAACAGTTCAACTATAAAGACCCAGTTAACGGTGTTGACATTGCTTAC ATCAAAATCCCGAACGCTGGCCAGATGCAGCCGGTAAAGGCATTCAAAATCCACAACAAAATCTGGGTT ATCCCGGAACGTGATACCTTTACTAACCCGGAAGAAGGTGACCTGAACCCGCCACCGGAAGCGAAACAG GTGCCGGTATCTTACTATGACTCCACCTACCTGTCTACCGATAACGAAAAGGACAACTACCTGAAAGGT GTTACTAAACTGTTCGAGCGTATTTACTCCACCGACCTGGGCCGTATGCTGCTGACTAGCATCGTTCGC GGTATCCCGTTCTGGGGCGGTTCTACCATCGATACCGAACTGAAAGTAATCGACACTAACTGCATCAAC GTTATTCAGCCGGACGGTTCCTATCGTTCCGAAGAACTGAACCTGGTGATCATCGGCCCGTCTGCTGAT ATCATCCAGTTCGAGTGTCTGAGCTTTGGTCACGAAGTTCTGAACCTCACCCGTAACGGCTACGGTTCC ACTCAGTACATCCGTTTCTCTCCGGACTTCACCTTCGGTTTTGAAGAATCCCTGGAAGTAGACACGAAC CCACTGCTGGGCGCTGGTAAATTCGCAACTGATCCTGCGGTTACCCTGGCTCACGAACTGATTCATGCA GGCCACCGCCTGTACGGTATCGCCATCAATCCGAACCGTGTCTTCAAAGTTAACACCAACGCGTATTAC GAGATGTCCGGTCTGGAAGTTAGCTTCGAAGAACTGCGTACTTTTGGCGGTCACGACGCTAAATTCATC GACTCTCTGCAAGAAAACGAGTTCCGTCTGTACTACTATAACAAGTTCAAAGATATCGCATCCACCCTG AACAAAGCGAAATCCATCGTGGGTACCACTGCTTCTCTCCAGTACATGAAGAACGTTTTTAAAGAAAAA TACCTGCTCAGCGAAGACACCTCCGGCAAATTCTCTGTAGACAAGTTGAAATTCGATAAACTTTACAAA ATGCTGACTGAAATTTACACCGAAGACAACTTCGTTAAGTTCTTTAAAGTTCTGAACCGCAAAACCTAT CTGAACTTCGACAAGGCAGTATTCAAAATCAACATCGTGCCGAAAGTTAACTACACTATCTACGATGGT TTCAACCTGCGTAACACCAACCTGGCTGCTAATTTTAACGGCCAGAACACGGAAATCAACAACATGAAC TTCACAAAACTGAAAAACTTCACTGGTCTGTTCGAGTTTTACAAGCTGCTGTGCGTCGACGGCATCATT ACCTCCAAAACTAAATCTGACGATGACGATAAAAACAAAGCGCTGAACCTGCAGTGTATCAAGGTTAAC AACTGGGATTTATTCTTCAGCCCGAGTGAAGACAACTTCACCAACGACCTGAACAAAGGTGAAGAAATC ACCTCAGATACTAACATCGAAGCAGCCGAAGAAAACATCTCGCTGGACCTGATCCAGCAGTACTACCTG ACCTTTAATTTCGACAACGAGCCGGAAAACATTTCTATCGAAAACCTGAGCTCTGATATCATCGGCCAG CTGGAACTGATGCCGAACATCGAACGTTTCCCAAACGGTAAAAAGTACGAGCTGGACAAATATACCATG TTCCACTACCTGCGCGCGCAGGAATTTGAACACGGCAAATCCCGTATCGCACTGACTAACTCCGTTAAC GAAGCTCTGCTCAACCCGTCCCGTGTATACACCTTCTTCTCTAGCGACTACGTGAAAAAGGTCAACAAA GCGACTGAAGCTGCAATGTTCTTGGGTTGGGTTGAACAGCTTGTTTATGATTTTACCGACGAGACGTCC GAAGTATCTACTACCGACAAAATTGCGGATATCACTATCATCATCCCGTACATCGGTCCGGCTCTGAAC ATTGGCAACATGCTGTACAAAGACGACTTCGTTGGCGCACTGATCTTCTCCGGTGCGGTGATCCTGCTG GAGTTCATCCCGGAAATCGCCATCCCGGTACTGGGCACCTTTGCTCTGGTTTCTTACATTGCAAACAAG GTTCTGACTGTACAAACCATCGACAACGCGCTGAGCAAACGTAACGAAAAATGGGATGAAGTTTACAAA TATATCGTGACCAACTGGCTGGCTAAGGTTAATACTCAGATCGACCTCATCCGCAAAAAAATGAAAGAA GCACTGGAAAACCAGGCGGAAGCTACCAAGGCAATCATTAACTACCAGTACAACCAGTACACCGAGGAA GAAAAAAACAACATCAACTTCAACATCGACGATCTGTCCTCTAAACTGAACGAATCCATCAACAAAGCT ATGATCAACATCAACAAGTTCCTGAACCAGTGCTCTGTAAGCTATCTGATGAACTCCATGATCCCGTAC GGTGTTAAACGTCTGGAGGACTTCGATGCGTCTCTGAAAGACGCCCTGCTGAAATACATTTACGACAAC CGTGGCACTCTGATCGGTCAGGTTGATCGTCTGAAGGACAAAGTGAACAATACCTTATCGACCGACATC CCTTTTCAGCTCAGTAAATATGTCGATAACCAACGCCTTTTGTCCACTctagaataatgaaagctt 2.DNAsequenceofLH.sub.N/B GGATCCATGCCGGTTACCATCAACAACTTCAACTACAACGACCCGATCGACAACAACAACATCATTATG ATGGAACCGCCGTTCGCACGTGGTACCGGACGTTACTACAAGGCTTTTAAGATCACCGACCGTATCTGG ATCATCCCGGAACGTTACACCTTCGGTTACAAACCTGAGGACTTCAACAAGAGTAGCGGGATTTTCAAT CGTGACGTCTGCGAGTACTATGATCCAGATTATCTGAATACCAACGATAAGAAGAACATATTCCTTCAG ACTATGATTAAACTCTTCAACCGTATCAAAAGCAAACCGCTCGGTGAAAAACTCCTCGAAATGATTATC AACGGTATCCCGTACCTCGGTGACCGTCGTGTCCCGCTTGAAGAGTTCAACACCAACATCGCAAGCGTC ACCGTCAACAAACTCATCAGCAACCCAGGTGAAGTCGAACGTAAAAAAGGTATCTTCGCAAACCTCATC ATCTTCGGTCCGGGTCCGGTCCTCAACGAAAACGAAACCATCGACATCGGTATCCAGAACCACTTCGCA AGCCGTGAAGGTTTCGGTGGTATCATGCAGATGAAATTCTGCCCGGAATACGTCAGTGTCTTCAACAAC GTCCAGGAAAACAAAGGTGCAAGCATCTTCAACCGTCGTGGTTACTTCAGCGACCCGGCACTCATCCTC ATGCATGAACTCATCCACGTCCTCCACGGTCTCTACGGTATCAAAGTTGACGACCTCCCGATCGTCCCG AACGAGAAGAAATTCTTCATGCAGAGCACCGACGCAATCCAGGCTGAGGAACTCTACACCTTCGGTGGC CAAGACCCAAGTATCATAACCCCGTCCACCGACAAAAGCATCTACGACAAAGTCCTCCAGAACTTCAGG GGTATCGTGGACAGACTCAACAAAGTCCTCGTCTGCATCAGCGACCCGAACATCAATATCAACATATAC AAGAACAAGTTCAAAGACAAGTACAAATTCGTCGAGGACAGCGAAGGCAAATACAGCATCGACGTAGAA AGTTTCGACAAGCTCTACAAAAGCCTCATGTTCGGTTTCACCGAAACCAACATCGCCGAGAACTACAAG ATCAAGACAAGGGCAAGTTACTTCAGCGACAGCCTCCCGCCTGTCAAAATCAAGAACCTCTTAGACAAC GAGATTTACACAATTGAAGAGGGCTTCAACATCAGTGACAAAGACATGGAGAAGGAATACAGAGGTCAG AACAAGGCTATCAACAAACAGGCATACGAGGAGATCAGCAAAGAACACCTCGCAGTCTACAAGATCCAG ATGTGCGTCGACGGCATCATTACCTCCAAAACTAAATCTGACGATGACGATAAAAACAAAGCGCTGAAC CTGCAGTGCATCGACGTTGACAACGAAGACCTGTTCTTCATCGCTGACAAAAACAGCTTCAGTGACGAC CTGAGCAAAAACGAACGTATCGAATACAACACCCAGAGCAACTACATCGAAAACGACTTCCCGATCAAC GAACTGATCCTGGACACCGACCTGATAAGTAAAATCGAACTGCCGAGCGAAAACACCGAAAGTCTGACC GACTTCAACGTTGACGTTCCGGTTTACGAAAAACAGCCGGCTATCAAGAAAATCTTCACCGACGAAAAC ACCATCTTCCAGTACCTGTACAGCCAGACCTTCCCGCTGGACATCCGTGACATCAGTCTGACCAGCAGT TTCGACGACGCTCTGCTGTTCAGCAACAAAGTTTACAGTTTCTTCAGCATGGACTACATCAAAACCGCT AACAAAGTTGTTGAAGCAGGGCTGTTCGCTGGTTGGGTTAAACAGATCGTTAACGACTTCGTTATCGAA GCTAACAAAAGCAACACTATGGACAAAATCGCTGACATCAGTCTGATCGTTCCGTACATCGGTCTGGCT CTGAACGTTGGTAACGAAACCGCTAAAGGTAACTTTGAAAACGCTTTCGAGATCGCTGGTGCAAGCATC CTGCTGGAGTTCATCCCGGAACTGCTGATCCCGGTTGTTGGTGCTTTCCTGCTGGAAAGTTACATCGAC AACAAAAACAAGATCATCAAAACCATCGACAACGCTCTGACCAAACGTAACGAAAAATGGAGTGATATG TACGGTCTGATCGTTGCTCAGTGGCTGAGCACCGTCAACACCCAGTTCTACACCATCAAAGAAGGTATG TACAAAGCTCTGAACTACCAGGCTCAGGCTCTGGAAGAGATCATCAAATACCGTTACAACATCTACAGT GAGAAGGAAAAGAGTAACATCAACATCGACTTCAACGACATCAACAGCAAACTGAACGAAGGTATCAAC CAGGCTATCGACAACATCAACAACTTCATCAACGGTTGCAGTGTTAGCTACCTGATGAAGAAGATGATC CCGCTGGCTGTTGAAAAACTGCTGGACTTCGACAACACCCTGAAAAAGAACCTGCTGAACTACATCGAC GAAAACAAGCTGTACCTGATCGGTAGTGCTGAATACGAAAAAAGTAAAGTGAACAAATACCTGAAGACC ATCATGCCGTTCGACCTGAGTATCTACACCAACGACACCATCCTGATCGAAATGTTCAACAAATACAAC TCtctagaataatgaaagctt 3.DNAsequenceofLH.sub.N/C ggatccATGCCGATCACCATCAACAACTTCAACTACAGCGATCCGGTGGATAACAAAAACATCCTGTAC CTGGATACCCATCTGAATACCCTGGCGAACGAACCGGAAAAAGCGTTTCGTATCACCGGCAACATTTGG GTTATTCCGGATCGTTTTAGCCGTAACAGCAACCCGAATCTGAATAAACCGCCGCGTGTTACCAGCCCG AAAAGCGGTTATTACGATCCGAACTATCTGAGCACCGATAGCGATAAAGATACCTTCCTGAAAGAAATC ATCAAACTGTTCAAACGCATCAACAGCCGTGAAATTGGCGAAGAACTGATCTATCGCCTGAGCACCGAT ATTCCGTTTCCGGGCAACAACAACACCCCGATCAACACCTTTGATTTCGATGTGGATTTCAACAGCGTT GATGTTAAAACCCGCCAGGGTAACAATTGGGTGAAAACCGGCAGCATTAACCCGAGCGTGATTATTACC GGTCCGCGCGAAAACATTATTGATCCGGAAACCAGCACCTTTAAACTGACCAACAACACCTTTGCGGCG CAGGAAGGTTTTGGCGCGCTGAGCATTATTAGCATTAGCCCGCGCTTTATGCTGACCTATAGCAACGCG ACCAACGATGTTGGTGAAGGCCGTTTCAGCAAAAGCGAATTTTGCATGGACCCGATCCTGATCCTGATG CATGAACTGAACCATGCGATGCATAACCTGTATGGCATCGCGATTCCGAACGATCAGACCATTAGCAGC GTGACCAGCAACATCTTTTACAGCCAGTACAACGTGAAACTGGAATATGCGGAAATCTATGCGTTTGGC GGTCCGACCATTGATCTGATTCCGAAAAGCGCGCGCAAATACTTCGAAGAAAAAGCGCTGGATTACTAT CGCAGCATTGCGAAACGTCTGAACAGCATTACCACCGCGAATCCGAGCAGCTTCAACAAATATATCGGC GAATATAAACAGAAACTGATCCGCAAATATCGCTTTGTGGTGGAAAGCAGCGGCGAAGTTACCGTTAAC CGCAATAAATTCGTGGAACTGTACAACGAACTGACCCAGATCTTCACCGAATTTAACTATGCGAAAATC TATAACGTGCAGAACCGTAAAATCTACCTGAGCAACGTGTATACCCCGGTGACCGCGAATATTCTGGAT GATAACGTGTACGATATCCAGAACGGCTTTAACATCCCGAAAAGCAACCTGAACGTTCTGTTTATGGGC CAGAACCTGAGCCGTAATCCGGCGCTGCGTAAAGTGAACCCGGAAAACATGCTGTACCTGTTCACCAAA TTTTGCGTCGACGCGATTGATGGTCGTAGCCTGTACAACAAAACCCTGCAGTGTCGTGAACTGCTGGTG AAAAACACCGATCTGCCGTTTATTGGCGATATCAGCGATGTGAAAACCGATATCTTCCTGCGCAAAGAT ATCAACGAAGAAACCGAAGTGATCTACTACCCGGATAACGTGAGCGTTGATCAGGTGATCCTGAGCAAA AACACCAGCGAACATGGTCAGCTGGATCTGCTGTATCCGAGCATTGATAGCGAAAGCGAAATTCTGCCG GGCGAAAACCAGGTGTTTTACGATAACCGTACCCAGAACGTGGATTACCTGAACAGCTATTACTACCTG GAAAGCCAGAAACTGAGCGATAACGTGGAAGATTTTACCTTTACCCGCAGCATTGAAGAAGCGCTGGAT AACAGCGCGAAAGTTTACACCTATTTTCCGACCCTGGCGAACAAAGTTAATGCGGGTGTTCAGGGCGGT CTGTTTCTGATGTGGGCGAACGATGTGGTGGAAGATTTCACCACCAACATCCTGCGTAAAGATACCCTG GATAAAATCAGCGATGTTAGCGCGATTATTCCGTATATTGGTCCGGCGCTGAACATTAGCAATAGCGTG CGTCGTGGCAATTTTACCGAAGCGTTTGCGGTTACCGGTGTGACCATTCTGCTGGAAGCGTTTCCGGAA TTTACCATTCCGGCGCTGGGTGCGTTTGTGATCTATAGCAAAGTGCAGGAACGCAACGAAATCATCAAA ACCATCGATAACTGCCTGGAACAGCGTATTAAACGCTGGAAAGATAGCTATGAATGGATGATGGGCACC TGGCTGAGCCGTATTATCACCCAGTTCAACAACATCAGCTACCAGATGTACGATAGCCTGAACTATCAG GCGGGTGCGATTAAAGCGAAAATCGATCTGGAATACAAAAAATACAGCGGCAGCGATAAAGAAAACATC AAAAGCCAGGTTGAAAACCTGAAAAACAGCCTGGATGTGAAAATTAGCGAAGCGATGAATAACATCAAC AAATTCATCCGCGAATGCAGCGTGACCTACCTGTTCAAAAACATGCTGCCGAAAGTGATCGATGAACTG AACGAATTTGATCGCAACACCAAAGCGAAACTGATCAACCTGATCGATAGCCACAACATTATTCTGGTG GGCGAAGTGGATAAACTGAAAGCGAAAGTTAACAACAGCTTCCAGAACACCATCCCGTTTAACATCTTC AGCTATACCAACAACAGCCTGCTGAAAGATATCATCAACGAATACTTCAAtctagaataatgaaagctt 4.DNAsequenceofLH.sub.N/D ggatccATGACGTGGCCAGTTAAGGATTTCAACTACTCAGATCCTGTAAATGACAACGATATTCTGTAC CTTCGCATTCCACAAAATAAACTGATCACCACACCAGTCAAAGCATTCATGATTACTCAAAACATTTGG GTCATTCCAGAACGCTTTTCTAGTGACACAAATCCGAGTTTATCTAAACCTCCGCGTCCGACGTCCAAA TATCAGAGCTATTACGATCCCTCATATCTCAGTACGGACGAACAAAAAGATACTTTCCTTAAAGGTATC ATTAAACTGTTTAAGCGTATTAATGAGCGCGATATCGGGAAAAAGTTGATTAATTATCTTGTTGTGGGT TCCCCGTTCATGGGCGATAGCTCTACCCCCGAAGACACTTTTGATTTTACCCGTCATACGACAAACATC GCGGTAGAGAAGTTTGAGAACGGATCGTGGAAAGTCACAAACATCATTACACCTAGCGTCTTAATTTTT GGTCCGCTGCCAAACATCTTAGATTATACAGCCAGCCTGACTTTGCAGGGGCAACAGTCGAATCCGAGT TTCGAAGGTTTTGGTACCCTGAGCATTCTGAAAGTTGCCCCGGAATTTCTGCTCACTTTTTCAGATGTC ACCAGCAACCAGAGCTCAGCAGTATTAGGAAAGTCAATTTTTTGCATGGACCCGGTTATTGCACTGATG CACGAACTGACGCACTCTCTGCATCAACTGTATGGGATCAACATCCCCAGTGACAAACGTATTCGTCCC CAGGTGTCTGAAGGATTTTTCTCACAGGATGGGCCGAACGTCCAGTTCGAAGAGTTGTATACTTTCGGA GGCCTGGACGTAGAGATCATTCCCCAGATTGAGCGCAGTCAGCTGCGTGAGAAGGCATTGGGCCATTAT AAGGATATTGCAAAACGCCTGAATAACATTAACAAAACGATTCCATCTTCGTGGATCTCGAATATTGAT AAATATAAGAAAATTTTTAGCGAGAAATATAATTTTGATAAAGATAATACAGGTAACTTTGTGGTTAAC ATTGACAAATTCAACTCCCTTTACAGTGATTTGACGAATGTAATGAGCGAAGTTGTGTATAGTTCCCAA TACAACGTTAAGAATCGTACCCATTACTTCTCTCGTCACTACCTGCCGGTTTTCGCGAACATCCTTGAC GATAATATTTACACTATTCGTGACGGCTTTAACTTGACCAACAAGGGCTTCAATATTGAAAATTCAGGC CAGAACATTGAACGCAACCCGGCCTTGCAGAAACTGTCGAGTGAATCCGTGGTTGACCTGTTTACCAAA GTCTGCGTCGACAAAAGCGAAGAGAAGCTGTACGATGACGATGACAAAGATCGTTGGGGATCGTCCCTG CAGTGTATTAAAGTGAAAAACAATCGGCTGCCTTATGTAGCAGATAAAGATAGCATTAGTCAGGAGATT TTCGAAAATAAAATTATCACTGACGAAACCAATGTTCAGAATTATTCAGATAAATTTTCACTGGACGAA AGCATCTTAGATGGCCAAGTTCCGATTAACCCGGAAATTGTTGATCCGTTACTGCCGAACGTGAATATG GAACCGTTAAACCTCCCTGGCGAAGAGATCGTATTTTATGATGACATTACGAAATATGTGGACTACCTT AATTCTTATTACTATTTGGAAAGCCAGAAACTGTCCAATAACGTGGAAAACATTACTCTGACCACAAGC GTGGAAGAGGCTTTAGGCTACTCAAATAAGATTTATACCTTCCTCCCGTCGCTGGCGGAAAAAGTAAAT AAAGGTGTGCAGGCTGGTCTGTTCCTCAACTGGGCGAATGAAGTTGTCGAAGACTTTACCACGAATATT ATGAAAAAGGATACCCTGGATAAAATCTCCGACGTCTCGGTTATTATCCCATATATTGGCCCTGCGTTA AATATCGGTAATAGTGCGCTGCGGGGGAATTTTAACCAGGCCTTTGCTACCGCGGGCGTCGCGTTCCTC CTGGAGGGCTTTCCTGAATTTACTATCCCGGCGCTCGGTGTTTTTACATTTTACTCTTCCATCCAGGAG CGTGAGAAAATTATCAAAACCATCGAAAACTGCCTGGAGCAGCGGGTGAAACGCTGGAAAGATTCTTAT CAATGGATGGTGTCAAACTGGTTATCTCGCATCACGACCCAATTCAACCATATTAATTACCAGATGTAT GATAGTCTGTCGTACCAAGCTGACGCCATTAAAGCCAAAATTGATCTGGAATATAAAAAGTACTCTGGT AGCGATAAGGAGAACATCAAAAGCCAGGTGGAGAACCTTAAGAATAGTCTGGATGTGAAAATCTCTGAA GCTATGAATAACATTAACAAATTCATTCGTGAATGTTCGGTGACGTACCTGTTCAAGAATATGCTGCCA AAAGTTATTGATGAACTGAATAAATTTGATCTGCGTACCAAAACCGAACTTATCAACCTCATCGACTCC CACAACATTATCCTTGTGGGCGAAGTGGATCGTCTGAAGGCCAAAGTAAACGAGAGCTTTGAAAATACG ATGCCGTTTAATATTTTTTCATATACCAATAACTCCTTGCTGAAAGATATCATCAATGAATATTTCAAT ctagattaataagctt 5.DNAsequenceoftheCT-CST29linker GGATCCGTCGACCTGCAGGGTCTAGAAGGCGGTGGCGGTAGCGGCGGTGGCGGTAGCGGCGGTGGCGGT AGCGGCGGTGGCGGTAGCGCACTAGTGCAGGAAAGACCTCCATTACAACAACCTCCACATCGCGATAAG AAACCATGTAAGAATTTCTTTTGGAAAACATTTAGCAGTTGCAAATGATAAAAGCTT 6.DNAsequenceoftheLHD-CT-CST29fusion GGATCCATGACGTGGCCAGTTAAGGATTTCAACTACTCAGATCCTGTAAATGACAACGATATTCTGTAC CTTCGCATTCCACAAAATAAACTGATCACCACACCAGTCAAAGCATTCATGATTACTCAAAACATTTGG GTCATTCCAGAACGCTTTTCTAGTGACACAAATCCGAGTTTATCTAAACCTCCGCGTCCGACGTCCAAA TATCAGAGCTATTACGATCCCTCATATCTCAGTACGGACGAACAAAAAGATACTTTCCTTAAAGGTATC ATTAAACTGTTTAAGCGTATTAATGAGCGCGATATCGGGAAAAAGTTGATTAATTATCTTGTTGTGGGT TCCCCGTTCATGGGCGATAGCTCTACCCCCGAAGACACTTTTGATTTTACCCGTCATACGACAAACATC GCGGTAGAGAAGTTTGAGAACGGATCGTGGAAAGTCACAAACATCATTACACCTAGCGTCTTAATTTTT GGTCCGCTGCCAAACATCTTAGATTATACAGCCAGCCTGACTTTGCAGGGGCAACAGTCGAATCCGAGT TTCGAAGGTTTTGGTACCCTGAGCATTCTGAAAGTTGCCCCGGAATTTCTGCTCACTTTTTCAGATGTC ACCAGCAACCAGAGCTCAGCAGTATTAGGAAAGTCAATTTTTTGCATGGACCCGGTTATTGCACTGATG CACGAACTGACGCACTCTCTGCATCAACTGTATGGGATCAACATCCCCAGTGACAAACGTATTCGTCCC CAGGTGTCTGAAGGATTTTTCTCACAGGATGGGCCGAACGTCCAGTTCGAAGAGTTGTATACTTTCGGA GGCCTGGACGTAGAGATCATTCCCCAGATTGAGCGCAGTCAGCTGCGTGAGAAGGCATTGGGCCATTAT AAGGATATTGCAAAACGCCTGAATAACATTAACAAAACGATTCCATCTTCGTGGATCTCGAATATTGAT AAATATAAGAAAATTTTTAGCGAGAAATATAATTTTGATAAAGATAATACAGGTAACTTTGTGGTTAAC ATTGACAAATTCAACTCCCTTTACAGTGATTTGACGAATGTAATGAGCGAAGTTGTGTATAGTTCCCAA TACAACGTTAAGAATCGTACCCATTACTTCTCTCGTCACTACCTGCCGGTTTTCGCGAACATCCTTGAC GATAATATTTACACTATTCGTGACGGCTTTAACTTGACCAACAAGGGCTTCAATATTGAAAATTCAGGC CAGAACATTGAACGCAACCCGGCCTTGCAGAAACTGTCGAGTGAATCCGTGGTTGACCTGTTTACCAAA GTCTGCGTCGACAAAAGCGAAGAGAAGCTGTACGATGACGATGACAAAGATCGTTGGGGATCGTCCCTG CAGTGTATTAAAGTGAAAAACAATCGGCTGCCTTATGTAGCAGATAAAGATAGCATTAGTCAGGAGATT TTCGAAAATAAAATTATCACTGACGAAACCAATGTTCAGAATTATTCAGATAAATTTTCACTGGACGAA AGCATCTTAGATGGCCAAGTTCCGATTAACCCGGAAATTGTTGATCCGTTACTGCCGAACGTGAATATG GAACCGTTAAACCTCCCTGGCGAAGAGATCGTATTTTATGATGACATTACGAAATATGTGGACTACCTT AATTCTTATTACTATTTGGAAAGCCAGAAACTGTCCAATAACGTGGAAAACATTACTCTGACCACAAGC GTGGAAGAGGCTTTAGGCTACTCAAATAAGATTTATACCTTCCTCCCGTCGCTGGCGGAAAAAGTAAAT AAAGGTGTGCAGGCTGGTCTGTTCCTCAACTGGGCGAATGAAGTTGTCGAAGACTTTACCACGAATATT ATGAAAAAGGATACCCTGGATAAAATCTCCGACGTCTCGGTTATTATCCCATATATTGGCCCTGCGTTA AATATCGGTAATAGTGCGCTGCGGGGGAATTTTAACCAGGCCTTTGCTACCGCGGGCGTCGCGTTCCTC CTGGAGGGCTTTCCTGAATTTACTATCCCGGCGCTCGGTGTTTTTACATTTTACTCTTCCATCCAGGAG CGTGAGAAAATTATCAAAACCATCGAAAACTGCCTGGAGCAGCGGGTGAAACGCTGGAAAGATTCTTAT CAATGGATGGTGTCAAACTGGTTATCTCGCATCACGACCCAATTCAACCATATTAATTACCAGATGTAT GATAGTCTGTCGTACCAAGCTGACGCCATTAAAGCCAAAATTGATCTGGAATATAAAAAGTACTCTGGT AGCGATAAGGAGAACATCAAAAGCCAGGTGGAGAACCTTAAGAATAGTCTGGATGTGAAAATCTCTGAA GCTATGAATAACATTAACAAATTCATTCGTGAATGTTCGGTGACGTACCTGTTCAAGAATATGCTGCCA AAAGTTATTGATGAACTGAATAAATTTGATCTGCGTACCAAAACCGAACTTATCAACCTCATCGACTCC CACAACATTATCCTTGTGGGCGAAGTGGATCGTCTGAAGGCCAAAGTAAACGAGAGCTTTGAAAATACG ATGCCGTTTAATATTTTTTCATATACCAATAACTCCTTGCTGAAAGATATCATCAATGAATATTTCAAT CTAGAAGGCGGTGGCGGTAGCGGCGGTGGCGGTAGCGGCGGTGGCGGTAGCGCACTAGTGCAGGAAAGA CCTCCATTACAACAACCTCCACATCGCGATAAGAAACCATGTAAGAATTTCTTTTGGAAAACATTTAGC AGTTGCAAAtaataagctt 7.ProteinsequenceoftheLHD-CT-CST29fusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDK FNSLYSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVQERPPL QQPPHRDKKPCKNFFWKTFSSCK 8.DNAsequenceoftheCP-EGFlinker ggatccGTCGACaacaacaataacaacaacaataacaacaacgacgatgacgataaaAATTCAGATAGC GAATGTCCACTTAGTCACGACGGGTACTGTTTGCATGATGGTGTGTGTATGTATATAGAAGCACTAGAC AAATACGCTTGCAATTGCGTAGTTGGCTATATAGGAGAGCGATGCCAATATAGAGATCTGAAGTGGTGG GAGTTAAGGGCAgaagcggcagccaaagaagcagccgctaaggcgctgcagagtctagaataataagct t 9.DNAsequenceoftheLHA-CP-EGFfusion ggatccATGGAGTTCGTTAACAAACAGTTCAACTATAAAGACCCAGTTAACGGTGTTGACATTGCTTAC ATCAAAATCCCGAACGCTGGCCAGATGCAGCCGGTAAAGGCATTCAAAATCCACAACAAAATCTGGGTT ATCCCGGAACGTGATACCTTTACTAACCCGGAAGAAGGTGACCTGAACCCGCCACCGGAAGCGAAACAG GTGCCGGTATCTTACTATGACTCCACCTACCTGTCTACCGATAACGAAAAGGACAACTACCTGAAAGGT GTTACTAAACTGTTCGAGCGTATTTACTCCACCGACCTGGGCCGTATGCTGCTGACTAGCATCGTTCGC GGTATCCCGTTCTGGGGCGGTTCTACCATCGATACCGAACTGAAAGTAATCGACACTAACTGCATCAAC GTTATTCAGCCGGACGGTTCCTATCGTTCCGAAGAACTGAACCTGGTGATCATCGGCCCGTCTGCTGAT ATCATCCAGTTCGAGTGTCTGAGCTTTGGTCACGAAGTTCTGAACCTCACCCGTAACGGCTACGGTTCC ACTCAGTACATCCGTTTCTCTCCGGACTTCACCTTCGGTTTTGAAGAATCCCTGGAAGTAGACACGAAC CCACTGCTGGGCGCTGGTAAATTCGCAACTGATCCTGCGGTTACCCTGGCTCACGAACTGATTCATGCA GGCCACCGCCTGTACGGTATCGCCATCAATCCGAACCGTGTCTTCAAAGTTAACACCAACGCGTATTAC GAGATGTCCGGTCTGGAAGTTAGCTTCGAAGAACTGCGTACTTTTGGCGGTCACGACGCTAAATTCATC GACTCTCTGCAAGAAAACGAGTTCCGTCTGTACTACTATAACAAGTTCAAAGATATCGCATCCACCCTG AACAAAGCGAAATCCATCGTGGGTACCACTGCTTCTCTCCAGTACATGAAGAACGTTTTTAAAGAAAAA TACCTGCTCAGCGAAGACACCTCCGGCAAATTCTCTGTAGACAAGTTGAAATTCGATAAACTTTACAAA ATGCTGACTGAAATTTACACCGAAGACAACTTCGTTAAGTTCTTTAAAGTTCTGAACCGCAAAACCTAT CTGAACTTCGACAAGGCAGTATTCAAAATCAACATCGTGCCGAAAGTTAACTACACTATCTACGATGGT TTCAACCTGCGTAACACCAACCTGGCTGCTAATTTTAACGGCCAGAACACGGAAATCAACAACATGAAC TTCACAAAACTGAAAAACTTCACTGGTCTGTTCGAGTTTTACAAGCTGCTGTGCGTCGACaacaacaat aacaacaacaataacaacaacgacgatgacgataaaAATTCAGATAGCGAATGTCCACTTAGTCACGAC GGGTACTGTTTGCATGATGGTGTGTGTATGTATATAGAAGCACTAGACAAATACGCTTGCAATTGCGTA GTTGGCTATATAGGAGAGCGATGCCAATATAGAGATCTGAAGTGGTGGGAGTTAAGGGCAgaagcggca gccaaagaagcagccgctaaggcgCTGCAGTGTATCAAGGTTAACAACTGGGATTTATTCTTCAGCCCG AGTGAAGACAACTTCACCAACGACCTGAACAAAGGTGAAGAAATCACCTCAGATACTAACATCGAAGCA GCCGAAGAAAACATCTCGCTGGACCTGATCCAGCAGTACTACCTGACCTTTAATTTCGACAACGAGCCG GAAAACATTTCTATCGAAAACCTGAGCTCTGATATCATCGGCCAGCTGGAACTGATGCCGAACATCGAA CGTTTCCCAAACGGTAAAAAGTACGAGCTGGACAAATATACCATGTTCCACTACCTGCGCGCGCAGGAA TTTGAACACGGCAAATCCCGTATCGCACTGACTAACTCCGTTAACGAAGCTCTGCTCAACCCGTCCCGT GTATACACCTTCTTCTCTAGCGACTACGTGAAAAAGGTCAACAAAGCGACTGAAGCTGCAATGTTCTTG GGTTGGGTTGAACAGCTTGTTTATGATTTTACCGACGAGACGTCCGAAGTATCTACTACCGACAAAATT GCGGATATCACTATCATCATCCCGTACATCGGTCCGGCTCTGAACATTGGCAACATGCTGTACAAAGAC GACTTCGTTGGCGCACTGATCTTCTCCGGTGCGGTGATCCTGCTGGAGTTCATCCCGGAAATCGCCATC CCGGTACTGGGCACCTTTGCTCTGGTTTCTTACATTGCAAACAAGGTTCTGACTGTACAAACCATCGAC AACGCGCTGAGCAAACGTAACGAAAAATGGGATGAAGTTTACAAATATATCGTGACCAACTGGCTGGCT AAGGTTAATACTCAGATCGACCTCATCCGCAAAAAAATGAAAGAAGCACTGGAAAACCAGGCGGAAGCT ACCAAGGCAATCATTAACTACCAGTACAACCAGTACACCGAGGAAGAAAAAAACAACATCAACTTCAAC ATCGACGATCTGTCCTCTAAACTGAACGAATCCATCAACAAAGCTATGATCAACATCAACAAGTTCCTG AACCAGTGCTCTGTAAGCTATCTGATGAACTCCATGATCCCGTACGGTGTTAAACGTCTGGAGGACTTC GATGCGTCTCTGAAAGACGCCCTGCTGAAATACATTTACGACAACCGTGGCACTCTGATCGGTCAGGTT GATCGTCTGAAGGACAAAGTGAACAATACCTTATCGACCGACATCCCTTTTCAGCTCAGTAAATATGTC GATAACCAACGCCTTTTGTCCACTctagaataatgaaagctt 10.ProteinsequenceoftheLHA-CP-EGFfusion EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDNNNNNNNNNNDDDDKNSDSECPLSHDGYCLHDGVCMYIEALDKYACNCVVGY IGERCQYRDLKWWELRAEAAAKEAAAKALQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSDTNIEAAEE NISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHYLRAQEFEH GKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVSTTDKIADI TIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNAL SKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDD LSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRL KDKVNNTLSTDIPFQLSKYVDNQRLLST 11.ProteinsequenceofLH.sub.N/A EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLST 12.ProteinsequenceofLH.sub.N/B PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDEEKLYDDDDKDRWGSSLQCIDVDNEDLFFIADKNSFSDDLSKNER IEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYL YSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANKSNT MDAIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLESYIDNKNKII KTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSN INIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENKLYL IGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNS 13.ProteinsequenceofLH.sub.N/C PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFN 14.ProteinsequenceofLH.sub.N/D TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDK FNSLYSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFN 15.SynthesisedGnRHpeptide pGlu-His-Trp-Ser-Tyr-Gly-Cys-Arg-Pro-Gly-NH2 16.ProteinsequenceoftheLHB-CT-SST28fusion PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDGIITSKTKSDDDDKNKALNLQCIDVDNEDLFFIADKNSFSDDLSK NERIEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIF QYLYSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANK SNTMDKIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLESYIDNKN KIIKTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKE KSNINIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENK LYLIGSAEYEKSKVNKYLKTIMPFDLSIYINDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALDSANSN PAMAPRERKAGCKNFFWKTFTSC 17.ProteinsequenceoftheLHA-CP-SST28fusion EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDIFINPEEGDLNPPPEAKQVPV SYYDSTYLSIDNEKDNYLKGVIKLFERIYSIDLGRMLLTSIVRGIPFWGGSTIDTELKVIDINCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRIFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGITASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNINLAANFNGQNTEINNMNFIK LKNFTGLFEFYKLLCVDGIITSKIKSDDDDKSANSNPAMAPRERKAGCKNFFWKIFTSCALAGGGGSGG GGSGGGGSALVLQCIKVNNWDLFFSPSEDNFINDLNKGEEITSDINIEAAEENISLDLIQQYYLIFNFD NEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHYLRAQEFEHGKSRIALINSVNEALLN PSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFIDETSEVSTIDKIADITIIIPYIGPALNIGNML YKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLIVQTIDNALSKRNEKWDEVYKYIVIN WLAKVNIQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESINKAMININ KFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGILIGQVDRLKDKVNNTLSTDIPFQLS KYVDNQRLLST 18.ProteinsequenceoftheLHD-CT-EGFfusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDINPSLSKPPRPTSKYQS YYDPSYLSTDEQKDIFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDIFDFTRHTTNIAVE KFENGSWKVINIIIPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLIFSDVISN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNIGNFVVNIDK FNSLYSDLINVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLINKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDEINVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLITSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DILDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFIFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRIKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVNSDSEC PLSHDGYCLHDGVCMYIEALDKYACNCVVGYIGERCQYRDLKWWELR 19.ProteinsequenceoftheLHD-CT-VIPfusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDINPSLSKPPRPTSKYQS YYDPSYLSTDEQKDIFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDIFDFTRHTTNIAVE KFENGSWKVINIIIPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLIFSDVISN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNIGNFVVNIDK FNSLYSDLINVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLINKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDEINVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLITSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DILDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFIFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRIKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVHSDAVF TDNYTRLRKQMAVKKYLNSILN 20.ProteinsequenceoftheLHC-CT-IGF1fusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVISPKSG YYDPNYLSTDSDKDIFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKIGSINPSVIITGPRENIIDPETSTFKLINNTFAAQEGFGALSIISISPRFMLTYSNAIND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVISNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVIVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNIDLPFIGDISDVKIDIFLRKDINE ETEVIYYPDNVSVDQVILSKNISEHGQLDLLYPSIDSESEILPGENQVFYDNRIQNVDYLNSYYYLESQ KLSDNVEDFIFIRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDILDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNIKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVGPETLCGAELVD ALQFVCGDRGFYFNKPIGYGSSSRRAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA 21.ProteinsequenceoftheLHD-CT-IGF1fusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDINPSLSKPPRPTSKYQS YYDPSYLSTDEQKDIFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDIFDFTRHTTNIAVE KFENGSWKVINIIIPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLIFSDVISN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNIGNFVVNIDK FNSLYSDLINVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLINKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDEINVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLITSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DILDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFIFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRIKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVGPETLC GAELVDALQFVCGDRGFYFNKPIGYGSSSRRAPQTGIVDECCFRSCDLRRLEMYCAPLKPAKSA 22.ProteinsequenceoftheLHC-CT-VIPfusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVISPKSG YYDPNYLSTDSDKDIFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKIGSINPSVIITGPRENIIDPETSTFKLINNTFAAQEGFGALSIISISPRFMLTYSNAIND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVISNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVIVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNIDLPFIGDISDVKIDIFLRKDINE ETEVIYYPDNVSVDQVILSKNISEHGQLDLLYPSIDSESEILPGENQVFYDNRIQNVDYLNSYYYLESQ KLSDNVEDFIFIRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDILDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNIKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVHSDAVFIDNYTR LRKQMAVKKYLNSILN 23.ProteinsequenceoftheLHC-CT-GnRHfusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVISPKSG YYDPNYLSTDSDKDIFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKIGSINPSVIITGPRENIIDPETSTFKLINNTFAAQEGFGALSIISISPRFMLTYSNAIND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVISNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVIVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNIDLPFIGDISDVKIDIFLRKDINE ETEVIYYPDNVSVDQVILSKNISEHGQLDLLYPSIDSESEILPGENQVFYDNRIQNVDYLNSYYYLESQ KLSDNVEDFIFIRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDILDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNIKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVMKPIQKLLAGLI LLTWCVEGCSSQHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQRFECITHQPRSPLRDLKGALES LIEEETGQKKI 24.ProteinsequenceoftheLHD-CT-GnRHfusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDINPSLSKPPRPTSKYQS YYDPSYLSTDEQKDIFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDIFDFTRHTTNIAVE KFENGSWKVINIIIPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLIFSDVISN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNIGNFVVNIDK FNSLYSDLINVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLINKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDEINVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLITSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DILDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFIFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRIKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSGGGGSALVM KPIQKLLAGLILLTWCVEGCSSQHWSYGLRPGGKRDAENLIDSFQEIVKEVGQLAETQRFECITHQPRS PLRDLKGALESLIEEETGQKKI 25.ProteinsequenceoftheLHD-CT-GRPfusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDINPSLSKPPRPTSKYQS YYDPSYLSTDEQKDIFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDIFDFTRHTTNIAVE KFENGSWKVINIIIPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLIFSDVISN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNIGNFVVNIDK FNSLYSDLINVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLINKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDEINVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLITSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DILDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFIFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRIKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVGNHWAV GHLM 26.ProteinsequenceoftheLHB-CT-GRPfusion PVTINNFNYNDPIDNNNIIMMEPPFARGIGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNINDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNINIASVIVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIIIPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDEEKLYDDDDKDRWGSSLQCIDVDNEDLFFIADKNSFSDDLSKNER IEYNTQSNYIENDFPINELILDIDLISKIELPSENTESLIDFNVDVPVYEKQPAIKKIFTDENTIFQYL YSQTFPLDIRDISLISSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANKSNT MDAIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLESYIDNKNKII KTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSN INIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNILKKNLLNYIDENKLYL IGSAEYEKSKVNKYLKTIMPFDLSITENDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALVGNHWAVGH LM 27.ProteinsequenceoftheLHC-CT-LIFfusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVISPKSG YYDPNYLSTDSDKDIFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKIGSINPSVIITGPRENIIDPETSTFKLINNTFAAQEGFGALSIISISPRFMLTYSNAIND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVISNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVIVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNIDLPFIGDISDVKIDIFLRKDINE ETEVIYYPDNVSVDQVILSKNISEHGQLDLLYPSIDSESEILPGENQVFYDNRIQNVDYLNSYYYLESQ KLSDNVEDFIFIRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDILDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNIKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVSPLPITPVNATC AIRHPCHNNLMNQIRSQLAQLNGSANALFILYYTAQGEPFPNNLDKLCGPNVIDFPPFHANGTEKAKLV ELYRIVVYLGTSLGNITRDQKILNPSALSLHSKLNATADILRGLLSNVLCRLCSKYHVGHVDVTYGPDT SGKDVFQKKKLGCQLLGKYKQIIAVLAQAF 28.ProteinsequenceoftheLHB-CP-LIFfusion PVTINNFNYNDPIDNNNIIMMEPPFARGIGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNINDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNINIASVIVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIIIPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDNNNNNNNNNNDDDDKSPLPITPVNATCAIRHPCHNNLMNQIRSQL AQLNGSANALFILYYTAQGEPFPNNLDKLCGPNVIDFPPFHANGTEKAKLVELYRIVVYLGTSLGNITR DQKILNPSALSLHSKLNATADILRGLLSNVLCRLCSKYHVGHVDVTYGPDTSGKDVFQKKKLGCQLLGK YKQIIAVLAQAFAEAAAKEAAAKALQCIDVDNEDLFFIADKNSFSDDLSKNERIEYNTQSNYIENDFPI NELILDIDLISKIELPSENTESLIDFNVDVPVYEKQPAIKKIFTDENTIFQYLYSQTFPLDIRDISLTS SFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANKSNIMDKIADISLIVPYIGL ALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLESYIDNKNKIIKTIDNALTKRNEKWSD MYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSNINIDFNDINSKLNEGI NQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNILKKNLLNYIDENKLYLIGSAEYEKSKVNKYLK TIMPFDLSIYINDTILIEMFNKYNS 29.ProteinsequenceoftheLHC-CT-FGF1fusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVISPKSG YYDPNYLSTDSDKDIFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKIGSINPSVIITGPRENIIDPETSTFKLINNTFAAQEGFGALSIISISPRFMLTYSNAIND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVISNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVIVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNIDLPFIGDISDVKIDIFLRKDINE ETEVIYYPDNVSVDQVILSKNISEHGQLDLLYPSIDSESEILPGENQVFYDNRIQNVDYLNSYYYLESQ KLSDNVEDFIFIRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDILDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNIKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALVMFNLPPGNYKKP KLLYCSNGGHFLRILPDGIVDGIRDRSDQHIQLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQTPN EECLFLERLEENHYNTYISKKHAEKNWFVGLKKNGSCKRGPRTHYGQKAILFLPLPVSSD 30.ProteinsequenceoftheLHA-CP-FGF1fusion EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDIFINPEEGDLNPPPEAKQVPV SYYDSTYLSIDNEKDNYLKGVIKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDINCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRITGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGITASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNINLAANFNGQNTEINNMNFIK LKNFTGLFEFYKLLCVDGIITSKIKSDDDDKMFNLPPGNYKKPKLLYCSNGGHFLRILPDGIVDGIRDR SDQHIQLQLSAESVGEVYIKSTETGQYLAMDITGLLYGSQTPNEECLFLERLEENHYNTYISKKHAEKN WFVGLKKNGSCKRGPRTHYGQKAILFLPLPVSSDGGGGSGGGGSGGGGSALVLQCIKVNNWDLFFSPSE DNFTNDLNKGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERF PNGKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGW VEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPV LGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATK AIINYQYNQYTEEEKNNINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDA SLKDALLKYIYDNRGTLIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLST 31.ProteinsequenceoftheLHA-CT-FGF9fusion EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTLEGGGGSGGGGSGGGGSGGGGSALVDHLGQSEA GGLPRGPAVTDLDHLKGILRRRQLYCRTGFHLEIFPNGTIQGTRKDHSRFGILEFISIAVGLVSIRGVD SGLYLGMNEKGELYGSEKLTQECVFREQFEENWYNTYSSNLYKHVDTGRRYYVALNKDGTPREGTRTKR HQKFTHFLPRPVDPDKVPELYKDILSQS 32.ProteinsequenceoftheLHC-CP-FGF9fusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDNNNNNNNNNNDDDDKDHLGQSEAGGLPRGPAVTDLDHLKGILRRR QLYCRTGFHLEIFPNGTIQGTRKDHSRFGILEFISIAVGLVSIRGVDSGLYLGMNEKGELYGSEKLTQE CVFREQFEENWYNTYSSNLYKHVDTGRRYYVALNKDGTPREGTRTKRHQKFTHFLPRPVDPDKVPELYK DILSQSAEAAAKEAAAKALQCRELLVKNTDLPFIGDISDVKTDIFLRKDINEETEVIYYPDNVSVDQVI LSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQKLSDNVEDFTFTRSIEE ALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDTLDKISDVSAIIPYIGPALNIS NSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTIDNCLEQRIKRWKDSYEWM MGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQVENLKNSLDVKISEAMN NINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEVDKLKAKVNNSFQNTIPF NIFSYTNNSLLKDIINEYFN 33.ProteinsequenceoftheIgA-HNtet-CT-SST14Fusion ESNQPEKNGTATKPENSGNTTSENGQTEPEKKLELRNVSDIELYSQTNGTYRQHVSLDGIPENTDTYFV KVKSSAFKDVYIPVASITEEKRNGQSVYKITAKAEKLQQELENKYVDNFTFYLDKKAKEENTNFTSFSN LVKAINQNPSGTYHLAASLNANEVELGPDERSYIKDTFTGRLIGEKDGKNYAIYNLKKPLFENLSGATV EKLSLKNVAISGKNDIGSLANEATNGTKIKQVHVDGCVDGIITSKTKSDDDDKNKALNLQCIKIKNEDL TFIAEKNSFSEEPFQDEIVSYNTKNKPLNFNYSLDKIIVDYNLQSKITLPNDRTTPVTKGIPYAPEYKS NAASTIEIHNIDDNTIYQYLYAQKSPTTLQRITMTNSVDDALINSTKIYSYFPSVISKVNQGAQGILFL QWVRDIIDDFTNESSQKTTIDKISDVSTIVPYIGPALNIVKQGYEGNFIGALETTGVVLLLEYIPEITL PVIAALSIAESSTQKEKIIKTIDNFLEKRYEKWIEVYKLVKAKWLGTVNTQFQKRSYQMYRSLEYQVDA IKKIIDYEYKIYSGPDKEQIADEINNLKNKLEEKANKAMININIFMRESSRSFLVNQMINEAKKQLLEF DTQSKNILMQYIKANSKFIGITELKKLESKINKVFSTPIPFSYSKNLDCWVDNEEDIDVLEGGGGSGGG GSGGGGSALVAGCKNFFWKTFTSC 34.ProteinsequenceoftheIgA-HNtet-CP-55T14fusion ESNQPEKNGTATKPENSGNTTSENGQTEPEKKLELRNVSDIELYSQTNGTYRQHVSLDGIPENTDTYFV KVKSSAFKDVYIPVASITEEKRNGQSVYKITAKAEKLQQELENKYVDNFTFYLDKKAKEENTNFTSFSN LVKAINQNPSGTYHLAASLNANEVELGPDERSYIKDTFTGRLIGEKDGKNYAIYNLKKPLFENLSGATV EKLSLKNVAISGKNDIGSLANEATNGTKIKQVHVDGCVDGIITSKTKSDDDDKAGCKNFFWKTFTSCAL AGGGGSGGGGSGGGGSALALQCIKIKNEDLTFIAEKNSFSEEPFQDEIVSYNTKNKPLNFNYSLDKIIV DYNLQSKITLPNDRTTPVTKGIPYAPEYKSNAASTIEIHNIDDNTIYQYLYAQKSPTTLQRITMTNSVD DALINSTKIYSYFPSVISKVNQGAQGILFLQWVRDIIDDFTNESSQKTTIDKISDVSTIVPYIGPALNI VKQGYEGNFIGALETTGVVLLLEYIPEITLPVIAALSIAESSTQKEKIIKTIDNFLEKRYEKWIEVYKL VKAKWLGTVNTQFQKRSYQMYRSLEYQVDAIKKIIDYEYKIYSGPDKEQIADEINNLKNKLEEKANKAM ININIFMRESSRSFLVNQMINEAKKQLLEFDTQSKNILMQYIKANSKFIGITELKKLESKINKVFSTPI PFSYSKNLDCWVDNEEDIDV 35.ProteinsequenceoftheLHA-CT-SST14fusion EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTLEGGGGSGGGGSGGGGSALVAGCKNFFWKTFTS C 36.ProteinsequenceoftheLHA-CT-EGFv3fusion EFVNKQFNYKDPVNGVDIAYIKIPNAGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPV SYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQ PDGSYRSEELNLVIIGPSADIIQFECLSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLL GAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSL QENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLT EIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTK LKNFTGLFEFYKLLCVDGIITSKTKSDDDDKNKALNLQCIKVNNWDLFFSPSEDNFTNDLNKGEEITSD TNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHY LRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVS TTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLT VQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKN NINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGT LIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTLEGGGGSGGGGSGGGGSGGGGSALVDNSDPKCP LSHEGYCLNDGVCMYIGTLDRYACNCVVGYVGERCQYRDLKLAELR 37.ProteinsequenceoftheLHE-CT-IL6fusion PKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTTPQDFHPPTSLKNGDSSYYD PNYLQSDEEKDRFLKIVTKIFNRINNNLSGGILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNG SQHILLPNVIIMGAEPDLFETNSSNISLRNNYMPSNHGFGSIAIVTFSPEYSFRFNDNSINEFIQDPAL TLMHELIHSLHGLYGAKGITTTCIITQQQNPLITNRKGINIEEFLTFGGNDLNIITVAQYNDIYTNLLN DYRKIASKLSKVQVSNPQLNPYKDIFQEKYGLDKDASGIYSVNINKFDDILKKLYSFTEFDLATKFQVK CRETYIGQYKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIIKPITGRGLVKKIIRFCVDG IITSKTKSLIEGRNKALNLQCIEINNGELFFVASENSYNDDNINTPKEIDDTVTSNNNYENDLDQVILN FNSESAPGLSDEKLNLTIQNDAYIPKYDSNGTSDIEQHDVNELNVFFYLDAQKVPEGENNVNLTSSIDT ALLEQPKIYTFFSSEFINNVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGLALNI GNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKNKVIKAINNALKERDEKWKEV YSFIVSNWMTKINTQFNKRKEQMYQALQNQVNAIKTIIESKYNSYTLEEKNELTNKYDIKQIENELNQK VSIAMNNIDRFLTESSISYLMKIINEVKINKLREYDENVKTYLLNYIIQHGSILGESQQELNSMVTDTL NNSIPFKLSSYTDDKILISYFNKFFKLEGGGGSGGGGSGGGGSGGGGSALVPPGEDSKDVAAPHRQPLT SSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIIT GLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWL QDMTTHLILRSFKEFLQSSLRALRQM 38.ProteinsequenceoftheLHB-CT-IL8fusion PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDGIITSKTKSLIEGRNKALNLQCIDVDNEDLFFIADKNSFSDDLSK NERIEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIF QYLYSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANK SNTMDKIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLESYIDNKN KIIKTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKE KSNINIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENK LYLIGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALVAKELR CQCIKTYSKPFHPKFIKELRVIESGPHCANTEIIVKLSDGRELCLDPKENWVQRVVEKFLKRAENS 39.ProteinsequenceoftheLHF-CP-GRAN4fusion PVAINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTNPSDFDPPASLKNGSS AYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGKVLLQEISYAKPYLGNDHTPIDEFSPVTRTTSVNI KLSTNVESSMLLNLLVLGAGPDIFESCCYPVRKLIDPDVVYDPSNYGFGSINIVTFSPEYEYTFNDISG GHNSSTESFIADPAISLAHELIHALHGLYGARGVTYEETIEVKQAPLMIAEKPIRLEEFLTFGGQDLNI ITSAMKEKIYNNLLANYEKIATRLSEVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYK KLYSFTESDLANKFKVKARNTYFIKYEFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQSIKLNPKIIDS IPDKGLVEKIVKFAVENNNNNNNNNNLGCVDGIITSKTKSLIEGRDVKCDMEVSCPDGYTCCRLQSGAW GCCPFTQAVCCEDHIHCCPAGFTCDTQKGTCEALAGGGGSGGGGSGGGGSALVLQCIEVNNSELFFVAS ESSYNENDINTPKEIDDTTNLNNNYENNLDEVILDYNSQTIPQISNIENLNTLVQDNSYVPEYDSNGTS EIEEYDVVDFNVFFYLHAQKVPEGETNISLTSSIDTALLEESKDIFFSSEFIDTINKPVNAALFIDWIS KVIRDFTTEATQKSTVDKIADISLIVPYVGLALNIIIEAEKGNFEEAFELLGVGILLEFVPELTIPVIL VFTIKSYIDSYENKNKAIKAINNSLIEREAKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQNQVDAI KTAIEYKYNNYTSDEKNRLESEYNINNIEEELNKKVSLAMKNIERFMTESSISYLMKLINEAKVGKLKK YDNHVKSDLLNYILDHRSILGEQTNELSDLVTSTLNSSIPFELSSYTNDKILIIYFNRLYKT 40.ProteinsequenceoftheLHD-CP-TGFafusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDK FNSLYSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDGGGGSGGGGSADDDDKVVSHFNDCPDSHTQFCFHGTCRFLVQEDK PACVCHSGYVGARCEHADLLALAGGGGSGGGGSGGGGSALALQCIKVKNNRLPYVADKDSISQEIFENK IITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSYY YLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKKD TLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREKI IKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDKE NIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNII LVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFN 41.ProteinsequenceoftheLHD-CP-TGFbfusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDK FNSLYSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDGGGGSGGGGSADDDDKALDTNYCFSSTEKNCCVRQLYIDFRKDLG WKWIHEPKGYHANFCLGPCPYIWSLDTQYSKVLALYNQHNPGASAAPCCVPQALEPLPIVYYVGRKPKV EQLSNMIVRSCKCSALAGGGGSGGGGSGGGGSALALQCIKVKNNRLPYVADKDSISQEIFENKIITDET NVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSYYYLESQK LSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKKDTLDKIS DVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREKIIKTIEN CLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDKENIKSQV ENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNIILVGEVD RLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFN 42.ProteinsequenceoftheLHB-CT-TNFafusion PVTINNFNYNDPIDNNNIIMMEPPFARGTGRYYKAFKITDRIWIIPERYTFGYKPEDFNKSSGIFNRDV CEYYDPDYLNTNDKKNIFLQTMIKLFNRIKSKPLGEKLLEMIINGIPYLGDRRVPLEEFNTNIASVTVN KLISNPGEVERKKGIFANLIIFGPGPVLNENETIDIGIQNHFASREGFGGIMQMKFCPEYVSVFNNVQE NKGASIFNRRGYFSDPALILMHELIHVLHGLYGIKVDDLPIVPNEKKFFMQSTDAIQAEELYTFGGQDP SIITPSTDKSIYDKVLQNFRGIVDRLNKVLVCISDPNININIYKNKFKDKYKFVEDSEGKYSIDVESFD KLYKSLMFGFTETNIAENYKIKTRASYFSDSLPPVKIKNLLDNEIYTIEEGFNISDKDMEKEYRGQNKA INKQAYEEISKEHLAVYKIQMCVDEEKLYDDDDKDRWGSSLQCIDVDNEDLFFIADKNSFSDDLSKNER IEYNTQSNYIENDFPINELILDTDLISKIELPSENTESLTDFNVDVPVYEKQPAIKKIFTDENTIFQYL YSQTFPLDIRDISLTSSFDDALLFSNKVYSFFSMDYIKTANKVVEAGLFAGWVKQIVNDFVIEANKSNT MDAIADISLIVPYIGLALNVGNETAKGNFENAFEIAGASILLEFIPELLIPVVGAFLLESYIDNKNKII KTIDNALTKRNEKWSDMYGLIVAQWLSTVNTQFYTIKEGMYKALNYQAQALEEIIKYRYNIYSEKEKSN INIDFNDINSKLNEGINQAIDNINNFINGCSVSYLMKKMIPLAVEKLLDFDNTLKKNLLNYIDENKLYL IGSAEYEKSKVNKYLKTIMPFDLSIYTNDTILIEMFNKYNSLEGGGGSGGGGSGGGGSALVRSSSRTPS DKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTH TISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAES GQVYFGIIAL 43.ProteinsequenceoftheLHD-CT-SDF1fusion TWPVKDFNYSDPVNDNDILYLRIPQNKLITTPVKAFMITQNIWVIPERFSSDTNPSLSKPPRPTSKYQS YYDPSYLSTDEQKDTFLKGIIKLFKRINERDIGKKLINYLVVGSPFMGDSSTPEDTFDFTRHTTNIAVE KFENGSWKVTNIITPSVLIFGPLPNILDYTASLTLQGQQSNPSFEGFGTLSILKVAPEFLLTFSDVTSN QSSAVLGKSIFCMDPVIALMHELTHSLHQLYGINIPSDKRIRPQVSEGFFSQDGPNVQFEELYTFGGLD VEIIPQIERSQLREKALGHYKDIAKRLNNINKTIPSSWISNIDKYKKIFSEKYNFDKDNTGNFVVNIDK FNSLYSDLTNVMSEVVYSSQYNVKNRTHYFSRHYLPVFANILDDNIYTIRDGFNLTNKGFNIENSGQNI ERNPALQKLSSESVVDLFTKVCVDKSEEKLYDDDDKDRWGSSLQCIKVKNNRLPYVADKDSISQEIFEN KIITDETNVQNYSDKFSLDESILDGQVPINPEIVDPLLPNVNMEPLNLPGEEIVFYDDITKYVDYLNSY YYLESQKLSNNVENITLTTSVEEALGYSNKIYTFLPSLAEKVNKGVQAGLFLNWANEVVEDFTTNIMKK DTLDKISDVSVIIPYIGPALNIGNSALRGNFNQAFATAGVAFLLEGFPEFTIPALGVFTFYSSIQEREK IIKTIENCLEQRVKRWKDSYQWMVSNWLSRITTQFNHINYQMYDSLSYQADAIKAKIDLEYKKYSGSDK ENIKSQVENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNKFDLRTKTELINLIDSHNI ILVGEVDRLKAKVNESFENTMPFNIFSYTNNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALKPVSLSY RCPCRFFESHVARANVKHLKILNTPNCALQIVARLKNNNRQVCIDPKLKWIQEYLEKALNKRFKM 44.ProteinsequenceoftheLHC-CT-VEGFfusion PITINNFNYSDPVDNKNILYLDTHLNTLANEPEKAFRITGNIWVIPDRFSRNSNPNLNKPPRVTSPKSG YYDPNYLSTDSDKDTFLKEIIKLFKRINSREIGEELIYRLSTDIPFPGNNNTPINTFDFDVDFNSVDVK TRQGNNWVKTGSINPSVIITGPRENIIDPETSTFKLTNNTFAAQEGFGALSIISISPRFMLTYSNATND VGEGRFSKSEFCMDPILILMHELNHAMHNLYGIAIPNDQTISSVTSNIFYSQYNVKLEYAEIYAFGGPT IDLIPKSARKYFEEKALDYYRSIAKRLNSITTANPSSFNKYIGEYKQKLIRKYRFVVESSGEVTVNRNK FVELYNELTQIFTEFNYAKIYNVQNRKIYLSNVYTPVTANILDDNVYDIQNGFNIPKSNLNVLFMGQNL SRNPALRKVNPENMLYLFTKFCVDAIDGRSLYNKTLQCRELLVKNTDLPFIGDISDVKTDIFLRKDINE ETEVIYYPDNVSVDQVILSKNTSEHGQLDLLYPSIDSESEILPGENQVFYDNRTQNVDYLNSYYYLESQ KLSDNVEDFTFTRSIEEALDNSAKVYTYFPTLANKVNAGVQGGLFLMWANDVVEDFTTNILRKDTLDKI SDVSAIIPYIGPALNISNSVRRGNFTEAFAVTGVTILLEAFPEFTIPALGAFVIYSKVQERNEIIKTID NCLEQRIKRWKDSYEWMMGTWLSRIITQFNNISYQMYDSLNYQAGAIKAKIDLEYKKYSGSDKENIKSQ VENLKNSLDVKISEAMNNINKFIRECSVTYLFKNMLPKVIDELNEFDRNTKAKLINLIDSHNIILVGEV DKLKAKVNNSFQNTIPFNIFSYINNSLLKDIINEYFNLEGGGGSGGGGSGGGGSALAPMAEGGGQNHHE VVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMR IKPHQGQHIGEMSFLQHNKCECRPKKDRARQEKKSVRGKGKGQKRKRKKSRYKSWSVYVGARCCLMPWS LPGPHPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLELNERTCRCDKPRR