RU2812113C2 - ANTIBODIES AGAINST GUCY2c AND THEIR USE - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: following is presented: an antibody that specifically binds to guanylate cyclase C (GUCY2c), bispecific antibodies that specifically bind to GUCY2c and CD3, methods of treating GUCY2c-related diseases in a patient, nucleic acid molecules, expression vectors containing a polynucleotide, host cells for obtaining antibodies and compositions for the treatment of diseases associated with GUCY2c. The methods of producing the antibody and bispecific antibody are also disclosed.

EFFECT: obtaining new antibodies with high affinity for GUCY2 and CD3.

36 cl, 38 dwg, 49 tbl, 35 ex

Description

LIST OF SEQUENCES

This application is filed electronically using EFS-Web and includes an electronically filed sequence listing in .txt format. The .txt file contains a sequence list named "PC72377-PRV2_Sequence_Listing_ST25_05142019.txt" created on May 14, 2019 and is 738 KB in size. The sequence listing contained in this .txt file is part of the description and is incorporated herein by reference in its entirety.

Field of invention

The present invention relates to antibodies, for example full-length antibodies or antigen-binding fragments thereof, that specifically bind to GUCY2c (guanylate cyclase C) and/or CD3 (cluster of differentiation 3). The present invention further relates to bispecific antibodies that specifically bind to CD3 and a tumor cell antigen (eg, bispecific antibodies that specifically bind to CD3 and GUCY2c). The present invention also relates to related molecules, for example, nucleic acids encoding such antibodies or bispecific antibodies, compositions and related methods, for example, methods for producing and purifying such antibodies and bispecific antibodies, and their use in diagnostics and therapy.

BACKGROUND OF THE ART

Malignancy is the leading cause of death worldwide, accounting for more than 7 million deaths each year. Mortality from malignant neoplasms is almost always associated with the spread of primary tumors to distant foci, forming metastases, which ultimately leads to death. This is especially true for gastrointestinal malignancies, including adenocarcinoma of the esophagus, stomach, colon, and rectum. Colorectal cancer (CRC) remains the fourth most diagnosed malignancy and the second leading cause of cancer death in the United States (Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin., 66:7-30, 2016). Worldwide, colorectal cancer accounts for 1.2 million new cases and 600,000 deaths per year (Brenner H, Kloor M, Pox CP. Colorectal cancer. Lancet, 383:1490-502, 2014).

Guanylate cyclase C (GUCY2c) (also known as STAR, ST receptor, GUC2C, GUCY2C, GC-C, and GCC) is a cell surface transmembrane receptor involved in the maintenance of intestinal fluid, electrolyte homeostasis, and cell proliferation (Carrithers et al., Proc Natl Acad Sci USA 100: 3018-3020, 2003; Mann et al., Biochem Biophys Res Commun 239: 463-466, 1997; Pitari et al., Proc Natl Acad Sci USA 100: 2695-2699, 2003); GenBank accession number NM.sub.-004963, and GenPept accession number NP-004954). This function is mediated by the binding of guanylin (Wiegand et al. FEBS Lett. 311:150-154, 1992) and uroguanylin (Hamra et al. Proc Natl Acad Sci USA 9(22):10464-10468, 1993). GUCY2c is also a receptor for thermostable enterotoxin (ST), a peptide produced by E. coli as well as other infectious organisms (Rao, MC Ciba Found. Symp. 112:74-93, 1985; Knoop FC and Owens, MJ Pharmacol. Toxicol Methods 28:67-72, 1992). ST binding to GUCY2c activates a signaling cascade leading to intestinal diseases such as diarrhea.

GUCY2c has been characterized as a protein involved in the development of malignancies, including colorectal cancer, pancreatic cancer, gastric cancer, liver cancer and esophageal cancer (Carrithers et al., Dis Colon Rectum 39:171-181, 1996; Buc et al. Eur J Cancer 41: 1618-1627, 2005; Carrithers et al., Gastroenterology 107: 1653-1661, 1994; Urbanski et al., Biochem Biophys Acta 1245: 29-36, 1995).

As a cell surface protein, GUCY2c can serve as a therapeutic target for receptor-binding proteins such as antibodies or ligands. GUCY2c is expressed on the apical side of the epithelial cells lining the mucosa of the small intestine, colon and rectum (Carrithers et al., Dis Colon Rectum 39: 171-181, 1996). GUCY2c expression persists after malignant transformation of intestinal epithelial cells, with expression in all primary and metastatic colorectal tumors (Carrithers et al., 1996; Buc et al.; Carrithers et al., 1994). GUCY2c expression has also been detected in esophageal cells diagnosed in Barrett's esophagus, esophageal cancer and gastric cancer.

There remains a need for molecules and/or compositions that can specifically target and specifically bind to metastatic colorectal cancer cells. There is a need for improved methods of treating individuals who are believed to be suffering from colorectal cancer, particularly individuals who are believed to be suffering from metastatic colorectal cancer cells.

Various strategies have been developed to generate bispecific antibodies that can recruit T cells to mediate tumor cell killing. Such bispecific antibodies can form a bridge between a tumor cell and an effector cell of the human immune system (NK cells, T cells, monocytes, macrophages or granulocytes), thereby allowing specific killing of the tumor cell. Although such bispecific antibodies have been proven to be preferable for therapeutic use, for example, for therapeutic concepts in the treatment of tumors, there remains a need for bispecific antibodies that will target GUCY2c-positive tumor cells with high activity and specificity. Such a molecule would be useful as a therapeutic agent in the case of gastrointestinal malignancies, in the treatment of malignancies and in the detection of GUCY2c on cells.

SUMMARY OF THE INVENTION

The present invention relates to antibodies, including bispecific antibodies, where the antibodies specifically bind to GUCY2c. The invention further relates to bispecific antibodies that can bind to GUCY2c and CD3.

In one aspect, the present invention provides an antibody that specifically binds guanylate cyclase C (GUCY2c), wherein the antibody comprises: (a) a heavy chain variable region (VH) comprising a complementarity determining region VH one (CDR1 VH), a complementarity determining region VH two (CDR2 VH) and complementarity determining region VH three (CDR3 VH) of the VH sequence shown in SEQ ID NO: 11, 19, 26, 33, 41, 48, 52, 57, 60, 62, 64, 65, 67, 69 , 71 or 73; and/or (b) a light chain variable region (VL) comprising a VL complementarity determining region one (VL CDR1), a VL complementarity determining region two (VL CDR2) and a VL complementarity determining region three (VL CDR3) of the VL sequence set forth in SEQ ID NO: 92, 100, 104, 106, 112, 119, 125, 129, 134, 136, 137, 138, 140, 143, 145, 147, 150, 152, 156, 158, 160, 162, 166, 170 , 171, 172, 173, 174 or 175.

In one embodiment, the present invention provides an antibody that specifically binds to GUCY2c, wherein the antibody comprises: (a) a VH region comprising (i) a VH CDR1 comprising the sequence SEQ ID NO: 12, 20, 27, 34, 42, 74 , 257, 258, 259, 260 or 261; (ii) CDR2 VH containing the sequence SEQ ID NO: 13, 21, 28, 35, 43, 53, 66, 68, 70, 72, 75, 262, 263, 264, 265, 266 or 267; and (iii) CDR3 VH containing the sequence SEQ ID NO: 14, 22, 29, 36 or 44; and/or (b) a VL region containing (i) a VL CDR1 containing the sequence SEQ ID NO: 93, 101, 105, 107, 113, 120, 148, 153, 163, or 167; (ii) CDR2 VL containing the sequence SEQ ID NO: 78, 94, 102, 108, 114, 141, 144, 146, 149, 151, 157, 159, 161, 164 or 168; and (iii) CDR3 VL containing the sequence SEQ ID NO: 95, 109, 115, 121, 142, 154, 165 or 169.

In one embodiment, the present invention provides an antibody wherein: (a) the VH region comprises (i) a VH CDR1 comprising the sequence SEQ ID NO: 74; (ii) CDR2 VH containing the sequence SEQ ID NO: 75; and (iii) CDR3 VH containing the sequence SEQ ID NO: 29; and/or (b) the VL region contains (i) a VL CDR1 containing the sequence of SEQ ID NO: 148; (ii) CDR2 VL containing the sequence SEQ ID NO: 149; and (iii) CDR3 VL containing the sequence SEQ ID NO: 142.

In another embodiment, the present invention provides an antibody wherein: (a) the VH region contains SEQ ID NO: 11, 19, 26, 33, 41, 48, 52, 57, 60, 62, 64, 65, 67, 69, 71 or 73; and/or (b) the VL region contains SEQ ID NO: 92, 100, 104, 106, 112, 119, 125, 129, 134, 136, 137, 138, 140, 143, 145, 147, 150, 152, 156 , 158, 160, 162, 166, 170, 171, 172, 173, 174 or 175.

In yet another embodiment, the present invention provides an antibody wherein the VH region comprises the sequence SEQ ID NO: 73; and wherein the VL region contains the sequence SEQ ID NO: 147.

In one embodiment, the present invention provides an antibody wherein the VH region comprises the sequence SEQ ID NO: 73 or a variant thereof with one or more conservative amino acid substitutions at residues not found in the VH region; and/or wherein the VL region contains the sequence SEQ ID NO: 147 or a variant thereof with one or more conservative amino acid substitutions in amino acids not found in the VL region.

In one embodiment, the present invention relates to an antibody that binds to GUCY2c and contains: CDR1 VL against GUCY2c containing amino acids having the sequence RASESV-X _L1.30 -X _L1.30a -YG-X _L1.30d -SLLQ; CDR2 VL against GUCY2c, containing amino acids having the sequence shown in SEQ ID NO: 149; CDR3 VL against GUCY2c, containing amino acids having the sequence shown in SEQ ID NO: 142; CDR1 VH against GUCY2c, containing amino acids having the sequence GFTFS-X _H1.31 -X _H1.32 -WMH; CDR2 VH against GUCY2c, containing amino acids having the sequence EIK- X _H2.52A -X _H2.53 -X _H2.54 -X _H2.55 -X _H2.56 -X _H2.57 -NVHEKFKD; and CDR3 VH against GUCY2c, containing amino acids having the sequence TX _H3.96 -X _H3.97 -X _H3.98 -X _H3.99 -X _H3.100 -GX _H3.100B -WF-X _H3.100E -X _{H3 .101} -V, where each of X _L1.30 , X _L1.30a , X _L1.30d , X H1.31, X _H1.32 , X _H2.52A , X _H2.53 , _{X H2.54 ,} X _{H2 .55} , X _H2.56 , X _H2.57 , X _H3.96 , X _H3.97 , X _H3.98 , X _H3.99 , X _H3.100 , X _H3.100B , X _H3.100E and X _{H3 .101} independently represents the amino acid residue from columns 4 and 5 of tables 42A and 42B.

In one aspect, X _L1.30 is D, N or S; X _L1.30a is Y, W or I; X _L1.30d is T, S or H; X _H1.31 is S, R, W, Y, A, H, P, Y, T, N, K, D, G or V; X _H1.32 is Y, R, L, T, K, P, I, N, M, V or S; X _H2.52A is P, T or V; X _H2.53 is S, A, L or R; X _H2.54 is N, T, R, H, K, M, S, A, Y, T or I; X _H2.55 is E, R, K, N, Y, G, L, A, M, S, H, D or Q; X _H2.56 is L, W, Y, F, V, I, N or H; X _H2.57 is T, M, S, L, N, Q or V; X _H3.96 is I, F or K; X _H3.97 is T, V, L, I, M, F, Y or A; X _H3.98 is T, N, R, G, L or I; X _H3.99 is T, K, L, W, A, S, M, P, N or R; X _H3.100 is E, G, A, H, S, D, T, R, Q, K, Y, L or M; X _H3.100B is Y or H; X _H3.100E is F or L; and X _H3.101 is D, Y, E or S.

In one embodiment, the present invention provides an antibody, wherein the antibody is selected from the group consisting of a Fab fragment, a Fab' fragment, an F(ab') ₂ fragment, an Fd fragment, an Fv fragment, a single chain Fv fragment ( scFv), disulfide bond stabilized Fv fragment (dsFv), single domain antibody (dAb), monoclonal antibody, chimeric antibody, bispecific antibody, trispecific antibody, polyspecific antibody, bispecific heterodimeric diabody, bispecific heterodimeric IgG, polyclonal antibody, labeled antibody, humanized antibody , human antibodies and their fragments.

In another embodiment, the antibody of the present invention further comprises a human or humanized VH scaffold and a human or humanized VL scaffold. In some embodiments, the VH framework comprises the sequence SEQ ID NO: 5, 6, 7, 8, 15, 16, 17, 18, 23, 24, 25, 30, 31, 32, 37, 38, 39, 40, 45, 46, 47, 49, 50, 51, 54, 55, 56, 58, 59, 61 or 63; and/or the VL framework contains the sequence SEQ ID NO: 80, 81, 82, 83, 86, 87, 88, 89, 96, 97, 98, 99, 103, 110, 111, 116, 117, 118, 122, 123 , 124, 126, 127, 128, 130, 131, 132, 133, 135, 139 or 155.

In another embodiment, the present invention provides an isolated human monoclonal antibody that binds to an epitope on the extracellular domain of GUCY2c, wherein the epitope comprises at least one amino acid residue selected from amino acid residues R73, S74, S75, T76, E78, G79, L80, L82 , L83, R84 or I86 SEQ ID NO: 406.

In one aspect, the epitope comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten amino acid residues selected from amino acid residues R73, S74, S75, T76, E78, G79, L80, L82, L83, R84 or I86 SEQ ID NO: 406. In another aspect, the epitope comprises amino acid residues R73, S74, S75, T76, E78 , G79, L80, L82, L83, R84, or I86 SEQ ID NO: 406. In a further aspect, the epitope comprises amino acids having the sequence set forth in SEQ ID NO: 406. In a further aspect, the epitope is a functional epitope.

In another aspect, the isolated human antibody and the amino acid contacts of the GUCY2c epitope are within 3.8 angstroms as determined by crystallography. For purposes of the invention, the term "within 3.8 angstroms" means that the contacts are 3.8 angstroms or less.

In one aspect, the antibody of the present invention is a bispecific antibody. In another aspect, the present invention provides a bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain.

In one embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-GUCY2c VL (anti-GUCY2c VL) and anti-CD3 VH (anti-CD3 VH), and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in a direction from N-terminus to C-terminus: (i) domain 2 containing anti-CD3 VL (anti-CD3 VL) and anti-GUCY2c VH (anti-GUCY2c VH), and (ii) a second heterodimerization promoting domain; where VL against GUCY2c and VH against GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

In another embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-CD3 VL and anti-GUCY2c VH, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-GUCY2c VL and anti-CD3 VH, and (ii) a second heterodimerization promoting domain; where VL against GUCY2c and VH against GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

In another embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-GUCY2c VH and anti-CD3 VL, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-CD3 VH and anti-GUCY2c VL, and (ii) a second heterodimerization promoting domain; where VH against GUCY2c and VL against GUCY2c form a domain that specifically binds to GUCY2c; and VH anti-CD3 and VL anti-CD3 form a domain that specifically binds to CD3.

In a further embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-CD3 VH and anti-GUCY2c VL, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain comprises, in a direction from N-terminus to C-terminus: (i) domain 2 containing anti-GUCY2c VH and anti-CD3 VL, and (ii) a second heterodimerization promoting domain, wherein anti-GUCY2c VL and anti-CD3 VH GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

In some embodiments of the above, each of the first heterodimerization promoting domain and the second heterodimerization promoting domain comprises a CH2 domain and a CH3 domain, wherein the amino acid sequence of each of the CH2 domain and/or the CH3 domain is modified to regulate heterodimerization and/or stabilize the bispecific antibody.

In some such embodiments, the amino acid sequence of the CH2 domain and/or the CH3 domain comprises at least one amino acid modification wherein: (a) the CH3 domain of the first heterodimerization promoting domain forms an overhang; and (b) the CH3 domain of the second heterodimerization promoting domain forms a depression.

In another such embodiment, the CH3 domain of the first heterodimerization promoting domain contains mutations Y349C and/or T366W; and the CH3 domain of the second heterodimerization promoting domain contains mutations S354C, T366S, L368A and/or Y407V (numbered according to the EU index).

In a specific embodiment described above, the first heterodimerization promoting domain contains the sequence SEQ ID NO: 188; wherein the second heterodimerization promoting domain contains the sequence SEQ ID NO: 189.

In additional embodiments, the anti-GUCY2c VL and the anti-CD3 VH are connected via a glycine-serine linker; and VL against CD3 and VH against GUCY2c are connected via a glycine-serine linker. In some such embodiments, the glycine-serine linker is linker 1 comprising the sequence SEQ ID NO: 190. In other embodiments, the glycine-serine linker is linker 2 containing the sequence SEQ ID NO: 191.

In additional embodiments of the above, domain 1 is covalently linked to the first heterodimerization promoting domain via a cysteine linker; and domain 2 is covalently linked to the second heterodimerization promoting domain via a cysteine linker; wherein the cysteine linker contains at least five amino acids. In some such embodiments, the cysteine linker is linker 3 comprising the sequence SEQ ID NO: 192.

In some embodiments, the first polypeptide chain is covalently linked to the second polypeptide chain through at least one disulfide bond. In some such embodiments, at least one disulfide bond is formed between linker 3 of the first polypeptide chain and linker 3 of the second polypeptide chain. In another such embodiment, at least one disulfide bond is formed between the first heterodimerization promoting domain and the second heterodimerization promoting domain. In specific embodiments, each disulfide bond is formed by linking two cysteine residues.

In additional embodiments of any of the above, the bispecific antibody of the present invention comprises (a) an anti-GUCY2c VH CDR1 comprising the sequence SEQ ID NO: 12, 20, 27, 34, 42, 74, 257, 258, 259, 260, or 261; (b) CDR2 VH against GUCY2c containing the sequence SEQ ID NO: 13, 21, 28, 35, 43, 53, 66, 68, 70, 72, 75, 262, 263, 264, 265, 266 or 267; (c) CDR3 VH against GUCY2c containing the sequence SEQ ID NO: 14, 22, 29, 36 or 44; (d) CDR1 VL against GUCY2c containing the sequence SEQ ID NO: 93, 101, 105, 107, 113, 120, 148, 153, 163 or 167; (e) CDR2 VL against GUCY2c containing the sequence SEQ ID NO: 78, 94, 102, 108, 114, 141, 144, 146, 149, 151, 157, 159, 161, 164 or 168; and (f) CDR3 VL against GUCY2c containing the sequence SEQ ID NO: 95, 109, 115, 121, 142, 154, 165 or 169.

In a specific embodiment, the bispecific antibody of the present invention comprises (a) an anti-GUCY2c VH CDR1 comprising the sequence SEQ ID NO: 74 or 259; (b) CDR2 VH against GUCY2c containing the sequence SEQ ID NO: 75 or 267; (c) CDR3 VH against GUCY2c containing the sequence SEQ ID NO: 29; (d) CDR1 VL against GUCY2c containing the sequence SEQ ID NO: 148; (e) CDR2 VL against GUCY2c containing the sequence SEQ ID NO: 149; and (f) CDR3 VL against GUCY2c containing the sequence SEQ ID NO: 142.

In specific embodiments, the bispecific antibody comprises: (a) an anti-GUCY2c VH region comprising the sequence SEQ ID NO: 73; and (b) an anti-GUCY2c VL region containing the sequence shown in SEQ ID NO: 147.

In additional embodiments, the present invention provides bispecific antibodies comprising (a) an anti-CD3 VH CDR1 comprising the sequence SEQ ID NO: 2, 268, or 277; (b) CDR2 VH anti-CD3 containing the sequence SEQ ID NO: 3, 10, 269 or 270; (c) CDR3 VH anti-CD3 containing the sequence SEQ ID NO: 4; (d) VL anti-CD3 CDR1 containing the sequence SEQ ID NO: 77, 85, 91, 278, 279 or 280; (e) CDR2 VL anti-CD3 containing SEQ ID NO: 78, or 281; and (f) an anti-CD3 VL CDR3 comprising the sequence SEQ ID NO: 79.

In a specific embodiment, the present invention provides a bispecific antibody comprising (a) an anti-CD3 VH CDR1 comprising the sequence SEQ ID NO: 2 or 268; (b) CDR2 VH anti-CD3 containing the sequence SEQ ID NO: 10 or 270; (c) CDR3 VH anti-CD3 containing the sequence SEQ ID NO: 4; (d) VL anti-CD3 CDR1 containing the sequence SEQ ID NO: 91; (e) VL anti-CD3 CDR2 containing the sequence SEQ ID NO: 78; and (f) an anti-CD3 VL CDR3 comprising the sequence SEQ ID NO: 79.

In one embodiment, the present invention provides a bispecific antibody comprising: (a) an anti-CD3 VH region comprising the sequence SEQ ID NO: 1, 9 or 273; and (b) an anti-CD3 VL region comprising the sequence SEQ ID NO: 76, 84, 90, 274, 275 or 276. In a specific embodiment, the bispecific antibody of the present invention comprises: (a) an anti-CD3 VH region comprising the sequence SEQ ID NO: 9; and (b) an anti-CD3 VL region comprising the sequence SEQ ID NO: 90.

In one embodiment, the present invention relates to a bispecific antibody comprising: CDR1 VL against GUCY2c, containing amino acids having the sequence RASESV-X _L1.30 -X _L1.30a -YG-X _L1.30d -SLLQ; CDR2 VL against GUCY2c, containing amino acids having the sequence shown in SEQ ID NO: 149; CDR3 VL against GUCY2c, containing amino acids having the sequence shown in SEQ ID NO: 142; CDR1 VH against GUCY2c, containing amino acids having the sequence GFTFS-X _H1.31 -X _H1.32 -WMH; CDR2 VH against GUCY2c, containing amino acids having the sequence EIK-X _H2.52A -X _H2.53 -X _H2.54 -X _H2.55 -X _H2.56 -X _H2.57 -NVHEKFKD; and CDR3 VH against GUCY2c, containing amino acids having the sequence TX _H3.96 -X _H3.97 -X _H3.98 -X _H3.99 -X _H3.100 -GX _H3.100B -WF-X _H3.100E -X _{H3 .101} -V, where each of X _L1.30 , X _L1.30a , X _L1.30d , X H1.31, X _H1.32 , X _H2.52A , X _H2.53 , _{X H2.54 ,} X _{H2 .55} , X _H2.56 , X _H2.57 , X _H3.96 , X _H3.97 , X _H3.98 , X _H3.99 , X _H3.100 , X _H3.100B , X _H3.100E and X _{H3 .101} independently represents an amino acid residue according to columns 4 and 5 of tables 42A and 42B.

In one aspect, X _L1.30 is D, N or S; X _L1.30a is Y, W or I; X _L1.30d is T, S or H; X _H1.31 is S, R, W, Y, A, H, P, Y, T, N, K, D, G or V; X _H1.32 is Y, R, L, T, K, P, I, N, M, V or S; X _H2.52A is P, T or V; X _H2.53 is S, A, L or R; X _H2.54 is N, T, R, H, K, M, S, A, Y, T or I; X _H2.55 is E, R, K, N, Y, G, L, A, M, S, H, D or Q; X _H2.56 is L, W, Y, F, V, I, N or H; X _H2.57 is T, M, S, L, N, Q or V; X _H3.96 is I, F or K; X _H3.97 is T, V, L, I, M, F, Y or A; X _H3.98 is T, N, R, G, L or I; X _H3.99 is T, K, L, W, A, S, M, P, N or R; X _H3.100 is E, G, A, H, S, D, T, R, Q, K, Y, L or M; X _H3.100B is Y or H; X _H3.100E is F or L; and X _H3.101 is D, Y, E or S.

In another embodiment, the present invention provides a bispecific antibody that binds to an epitope on the extracellular domain of GUCY2c, wherein the epitope contains at least one amino acid residue selected from amino acid residues R73, S74, S75, T76, E78, G79, L80, L82, L83 , R84 or I86 SEQ ID NO: 406.

In one aspect, the epitope comprises at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten amino acid residues selected from amino acid residues R73, S74, S75, T76, E78, G79, L80, L82, L83, R84 or I86 SEQ ID NO: 406. In another aspect, the epitope comprises amino acid residues R73, S74, S75, T76, E78 , G79, L80, L82, L83, R84, or I86 SEQ ID NO: 406. In a further aspect, the epitope comprises amino acids having the sequence set forth in SEQ ID NO: 406. In a further aspect, the epitope is a functional epitope.

In another aspect, the isolated human antibody and the amino acid contacts of the GUCY2c epitope are within 3.8 angstroms as determined by crystallography.

In one aspect, the present invention provides a bispecific antibody that specifically binds to GUCY2c and competes for binding with the bispecific antibody provided herein.

In one aspect, the present invention provides a bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain, and wherein: (a) the first polypeptide chain contains the following regions in the following order in the direction from N- end to C-terminus: VL anti-GUCY2c antibody (VL anti-GUCY2c) (SEQ ID NO: 147) - linker 1 (SEQ ID NO: 190) - VH anti-CD3 antibody (VH anti-CD3) (SEQ ID NO: 9) - linker 3 (SEQ ID NO: 192) - first heterodimerization promoting domain (SEQ ID NO: 188); and (b) the second polypeptide chain contains the following regions in the following order in the direction from N-terminus to C-terminus: anti-CD3 VL (anti-CD3 VL) (SEQ ID NO: 90) - linker 2 (SEQ ID NO: 191) - VH antibodies against GUCY2c (VH against GUCY2c) (SEQ ID NO: 73) - linker 3 (SEQ ID NO: 192) - second heterodimerization promoting domain (SEQ ID NO: 189); where VL against GUCY2c and VH against GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3; wherein the linker 3 of the first polypeptide chain and the linker 3 of the second polypeptide chain are covalently linked to each other by two disulfide bonds; wherein each of the first heterodimerization promoting domain and the second heterodimerization promoting domain comprises a CH2 domain and a CH3 domain; wherein the CH3 domain of the first heterodimerization promoting domain forms a protrusion, and the CH3 domain of the second heterodimerization promoting domain forms a recess; wherein at least one disulfide bond is formed between the CH3 domain of the first heterodimerization promoting domain and the CH3 domain of the second heterodimerization promoting domain.

In some embodiments, the present invention provides a bispecific antibody further comprising a human or humanized VH scaffold and a human or humanized VL scaffold. In some such embodiments, the bispecific antibody is a humanized antibody. In specific embodiments, the VH framework contains the sequence SEQ ID NO: 5, 6, 7, 8, 15, 16, 17, 18, 23, 24, 25, 30, 31, 32, 37, 38, 39, 40, 45, 46, 47, 49, 50, 51, 54, 55, 56, 58, 59, 61 or 63; and/or the VL framework contains the sequence SEQ ID NO: 80, 81, 82, 83, 86, 87, 88, 89, 96, 97, 98, 99, 103, 110, 111, 116, 117, 118, 122, 123 , 124, 126, 127, 128, 130, 131, 132, 133, 135, 139 or 155.

In one aspect, the bispecific antibody of the present invention specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain is produced by an expression vector with ATCC accession number PTA-124944 and the second polypeptide chain is produced by using an expression vector with ATCC registration number PTA-124943.

In specific embodiments, the antibody or bispecific antibody of the present invention (a) binds to the extracellular domain of human GUCY2c; (b) demonstrates an increased half-life in serum or tumor from 30 min to 100 days; and/or (c) exhibits a lower EC ₅₀ value of 0.0001 nM to 100 nM in the presence of increased levels of GUCY2c expression or increased levels of receptor density.

The present invention further relates to a pharmaceutical composition containing a therapeutically effective amount of an antibody or bispecific antibody provided herein and a pharmaceutically acceptable carrier.

In another aspect, the present invention provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient an anti-GUCY2c antibody of the present invention. The present invention also provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient a bispecific antibody of the present invention. The present invention further provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient a pharmaceutical composition containing an anti-GUCY2c antibody or a bispecific antibody as provided herein. In some such embodiments, the GUCY2c-associated disorder is a malignancy. In specific embodiments, the malignancy is a malignancy of the digestive system selected from the group consisting of malignancy of the esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile ducts, and pancreas.

The present invention further provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient a bispecific antibody as provided herein, or a pharmaceutical composition containing a bispecific antibody as provided herein, wherein the cytolytic T cell response is activated.

In one aspect, the present invention relates to an antibody, bispecific antibody, or pharmaceutical composition provided herein for use in therapy. The present invention further relates to an antibody or bispecific antibody provided herein for use in the manufacture of a medicament for use in therapy. In some embodiments, the therapy is a treatment for a GUCY2c-associated disorder. In specific embodiments, the GUCY2c-associated disorder is a malignancy. In some embodiments, the cancer is a digestive system cancer selected from the group consisting of cancer of the esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile duct, and pancreas. In specific embodiments, the therapy activates a cytolytic T cell response.

In one aspect, the present invention relates to a polynucleotide encoding an antibody or bispecific antibody as described herein. In another embodiment, the present invention relates to a vector containing the polynucleotides presented in the present description. In yet another embodiment, the present invention relates to a host cell containing the vectors presented in the present description. In some such embodiments, the host cell recombinantly produces an antibody or bispecific antibody provided herein. In specific embodiments, the host cell is selected from the group consisting of bacterial cell lines, mammalian cell lines, insect cell lines, and yeast cell lines. In a specific embodiment, the mammalian cell line is a CHO cell line. In one embodiment, the antibody or bispecific antibody is produced using a cell-free in vitro protein synthesis system.

In one aspect, the present invention relates to a method for producing an anti-GUCY2c antibody or a bispecific antibody as provided herein, comprising culturing a host cell under conditions leading to the production of an anti-GUCY2c antibody or a bispecific antibody as provided herein, and purifying the antibody or bispecific antibody. antibodies from the culture supernatant.

In another aspect, the present invention relates to the use of an anti-GUCY2c antibody, bispecific antibody, pharmaceutical composition, polynucleotide, vector or host cell provided herein in the manufacture of a medicament for the treatment of a GUCY2c-associated disorder.

In another embodiment, the present invention provides a composition comprising bispecific antibodies of the invention and a second therapeutic agent.

In another embodiment, the present invention provides a composition comprising bispecific antibodies of the invention and an antidiarrheal agent.

In one aspect, the antidiarrheal agent includes, but is not limited to, bismuth subgallate, Lactobacillus acidophilus , Saccharomyces boulardii , loperamide/simethicone, atropine/diphenoxylate, atropine/difenoxin, Saccharomyces boulardii lyophilisate, Lactobacillus acidophilus , loperamide, bismuth subsalicylate, Lactobacillus acidophilus / Lactobacillus bulgaricus , Lactobacillus rhamnosus , attapulgite, crofelemer, fluoroquinolone, antibiotic or octreotide.

In another aspect, the present invention relates to the use of a composition comprising a bispecific anti-GUCY2c antibody and an antidiarrheal agent as provided herein in the manufacture of a medicament for the treatment of a GUCY2c-associated disorder.

Other embodiments will be apparent from the following detailed description. Where aspects or embodiments of the invention are described in terms of the Markush group or other grouping of alternatives, the present invention relates not only to the entire group as a whole, but also to each member of the group individually and all possible subgroups of the main group, as well as to the main group , in which one or more members of the group are missing. The present invention also provides for the specific exclusion of one or more of any members of the groups in the invention described in the application.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1A, 1B, 1C and 1D show schematics of four alternative representations of anti-CD3-GUCY2c bispecific antibodies containing a first heterodimerization-promoting domain and a second heterodimerization-promoting domain containing an Fc chain optimized for knob-to-valley binding.

Figure 2 shows the binding of bispecific antibodies (A) GUCY2C-0247 and (B) GUCY2C-1608 to T84 tumor cells using a flow cytometry-based assay.

Figure 3 shows a flow cytometry-based assay to determine the binding of bispecific antibodies (A) GUCY2C-0247 and (B) GUCY2C-1608 to naïve human T cells.

Figure 4A shows GUCY2C-1608 and GUCY2C-0247 recruiting naïve human T cells to induce killing of T84 tumor cells. Figure 4B shows GUCY2C-1608-mediated cytotoxic T cell activity observed in the GUCY2c-expressing tumor cell line, LS1034. Figure 4C shows GUCY2C-1608-mediated cytotoxic T cell activity observed in the GUCY2c-expressing tumor cell line LS174T. Figure 4D shows that no activity was observed using GUCY2C-1608 in a GUCY2c-negative cell line.

Figures 5A-5F show in vitro analysis of cytokine release induced following GUCY2C-1608-mediated recruitment of naïve human T cells to GUCY2c-expressing T84 cells. Luminex-based assay revealed up-regulation of human IFN-gamma (Figure 5A), IL-10 (Figure 5B), IL-2 (Figure 5C), IL-4 (Figure 5D), IL-6 (Figure 5E) and TNF -alpha (Figure 5F).

Figure 6 shows the dose-dependent inhibition of tumor growth by GUCY2C-0247 in a tumor xenograft of the T84 colorectal carcinoma cell line in an adoptive transfer model.

Figure 7 shows dose-dependent inhibition of tumor growth by GUCY2C-0247 in a tumor xenograft of the colorectal carcinoma cell line HT55 in an adoptive transfer model.

Figure 8 shows dose-dependent inhibition of tumor growth by GUCY2C-1608 in a tumor xenograft of the colorectal carcinoma cell line HT55 in an adoptive transfer model.

Figure 9 shows dose-dependent inhibition of tumor growth by bispecific antibodies against GUCY2c, GUCY2C-0247 and GUCY2C-1608, in a tumor xenograft of the colorectal cancer cell line LS1034 in an adoptive transfer model.

Figure 10 shows inhibition of tumor growth by bispecific antibodies against GUCY2c, GUCY2C-0074, GUCY2C-0098 and GUCY2C-0105, in a tumor xenograft of the colorectal cancer cell line LS1034 in an adoptive transfer model.

Figure 11 shows the dose-dependent inhibition of tumor growth by GUCY2C-0098 in a colorectal carcinoma patient-derived tumor xenograft, PDX-CRX-11201, in an adoptive transfer model.

Figure 12 shows dose-dependent inhibition of tumor growth by GUCY2C-1608 in a colorectal carcinoma patient-derived tumor xenograft, PDX-CRX-11201, in an adoptive transfer model.

Figure 13 shows the inhibition of tumor growth by GUCY2C-0240 in a colorectal carcinoma patient-derived tumor xenograft, PDX-CRX-11201, in an adoptive transfer model.

Figure 14 shows inhibition of tumor growth by GUCY2C-0098 in colorectal carcinoma patient-derived tumor xenograft PDX-CRX-12213 in an adoptive transfer model.

Figure 15 shows inhibition of tumor growth by GUCY2C-0247 in a colorectal carcinoma patient-derived tumor xenograft, PDX-CRX-24225, in an adoptive transfer model.

Figure 16 shows the characterization of the cGMP-inducing and neutralizing abilities of the anti-GUCY2c-CD3 bispecific antibody in T84 cells.

Figure 17 shows LC/MS analysis of GUCY2C-1608 after purification.

Figure 18 shows the T cell-mediated cytotoxicity of the anti-GUCY2C bispecific antibody with anti-CD3 variants. Bispecific antibodies against GUCY2c with different CD3 variants recruit naïve human T cells to induce killing of T84 tumor cells.

Figure 19 shows a DELFIA ELISA competition assay for the ECD peptide GUCY2c.

Figure 20 shows a competitive DELFIA ELISA using the MS20 antibody.

Figure 21 shows details of the binding region between the GUCY2c peptide and the GUCY2C-1608 scFv peptide by crystallography.

Figure 22 shows the crystal structure of the two antigen binding sites on the anti-GUCY2c-CD3 bispecific antibody, showing that they are separated by approximately 70 Å and are on opposite sides of the molecule, directly opposite each other.

Figure 23 shows the results of combination studies using anti-VEGF antibody and axitinib, demonstrating additive tumor growth inhibition in combination with the bispecific anti-GUCY2c-CD3 antibody.

Figure 24 shows the results of combination studies using anti-PD1 antibody demonstrating additive tumor growth inhibition in combination with anti-GUCY2c-CD3 bispecific antibody.

Figure 25 shows the results of combination studies using anti-PD-L1 antibody demonstrating additive tumor growth inhibition in combination with anti-GUCY2c-CD3 bispecific antibody.

DETAILED DESCRIPTION OF THE INVENTION

The invention provided herein provides antibodies that specifically bind to GUCY2c (eg, human GUCY2c, mouse GUCY2c, rat GUCY2c, cynomolgus GUCY2c). In addition, the invention provided herein provides bispecific antibodies that specifically bind to CD3 (eg, human CD3) and tumor antigen (eg, GUCY2c). The invention also relates to polynucleotides encoding these antibodies, compositions containing these antibodies, and methods for producing and using these antibodies. The invention further provides methods for treating a condition associated with GUCY2c expression in an individual, such as a cancer, using antibodies (eg, an anti-GUCY2 antibody, an anti-CD3 antibody, or a bispecific antibody) as provided herein.

General methods

The practice of the present invention, unless otherwise indicated, uses generally accepted methods of molecular biology (including recombinant methods), microbiology, cell biology, biochemistry and immunology known to those skilled in the art. Such methods are fully described in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993-1998) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology (D. M. Weir and C. C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos, eds., 1987); Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction (Mullis et al., eds., 1994); Current Protocols in Immunology (J.E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D. Catty., ed., IRL Press, 1988-1989); Monoclonal antibodies: a practical approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using antibodies: a laboratory manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995). In some cases, terms with generally accepted understandable meaning is defined herein for clarity and/or reference, and the inclusion of such definitions herein should not necessarily be construed as substantially different from the meaning generally generally understood in the art.

The present invention will now be described in detail with reference to the following definitions and examples. All patents and publications, including all sequences described in such patents and publications, referred to herein are expressly incorporated by reference herein.

Definitions

In general, unless otherwise indicated, all technical terms, symbols and other scientific terms or terminology used in the present description should have the meaning commonly understood by specialists in the field to which the present invention relates. For example, the term "and/or" as used in a phrase such as "A and/or B" herein is intended to include A and B; A or B; A (separately); and B (separately). Likewise, the term “and/or” as used in a phrase such as “A, B and/or C” is intended to include each of the following embodiments: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (separately); B (separately) and C (separately).

As used herein, terms in the singular include the corresponding plural unless the context clearly indicates otherwise.

Within the scope of the invention, numeric ranges include numbers defining a range.

The term "about" or the like, when preceded by a list of numeric values or range, refers to each individual value in the list or range independently, as if each individual value in the list or range was immediately preceded by that term. Terms mean that the meanings to which they refer are exact, close or similar to them. For example, in some embodiments, the term "about" a particular value may mean a value that is 99%, 95%, or 90% of that value. For example, the expression "about 100" includes 99 and 101 and all values in between (eg, 99.1, 99.2, 99.3, 99.4, 99.5, etc.). As another example, if the temperature is 70°C, "about" 70°C could be 69°C, 66°C, or 63°C. It should be understood that these are examples only.

As used herein, the term "within 3.8 angstroms" means that the contacts are 3.8 angstroms or less, as determined by crystallography.

Within the scope of the invention, nucleic acids are written from left to right in the 5'-3' direction; amino acid sequences are written from left to right in amino- to carboxy-terminal orientation, respectively. Practitioners can find definitions and technical terms in Sambrook et al., 1989, and Ausubel FM et al., 1993. It should be understood that the present invention is not limited to the specific methodology, methods and reagents described, as they can be varied.

The terms "polypeptide", "oligopeptide", "peptide" and "protein" are used interchangeably herein to refer to amino acid chains of any length, preferably relatively short (eg, 10-100 amino acids). The chain may be linear or branched, may contain modified amino acids and/or may be interrupted by non-amino acids. The terms also include an amino acid chain modified naturally or by intervention; for example, through disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification such as conjugation to a tag moiety. Also included in the definition are, for example, polypeptides containing one or more amino acid analogues (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art. It should be understood that the polypeptides may be in the form of individual chains or associated chains. Preferably, mammalian polypeptides (polypeptides originally obtained from a mammalian body) are used, more preferably directly secreted into the medium.

An "antibody" is an immunoglobulin molecule capable of specifically binding to a target, such as a carbohydrate, polynucleotide, lipid, polypeptide, etc., through at least one antigen recognition site located in the variable region of the immunoglobulin molecule. As used herein, the term includes polyclonal antibody, monoclonal antibody, chimeric antibody, bispecific antibody, dual-specific antibody, bifunctional antibody, trispecific antibody, multispecific antibody, bispecific heterodimeric diabody, bispecific heterodimeric IgG, tagged antibody, humanized antibody, human antibody, and fragments thereof (such as Fab, Fab', F(ab') ₂ , Fv), single chain (ScFv) and domain antibodies (including, for example, shark and camelid antibodies), antibody-containing fusion proteins, and any other modified configuration of an immunoglobulin molecule, containing an antigen recognition site. An antibody includes an antibody of any class such as IgG, IgA or IgM (or a subclass thereof), and the antibody may not belong to any particular class. Depending on the amino acid sequence of the constant region of the heavy chains of antibodies, immunoglobulins can be assigned to different classes. There are five main classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and some of them can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant regions corresponding to different classes of immunoglobulins are named alpha, delta, epsilon, gamma and mu, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known. The present invention also includes “antibody analogues,” other non-antibody protein scaffolds, such as fusion proteins and/or immunoconjugates that use CDRs to promote specific antigen binding. The antibodies of the invention can be obtained from any species, including, but not limited to, mouse, human, camel, llama, fish, shark, goat, rabbit, chicken and cow.

The term "antibody" further includes immunoglobulin molecules containing four polypeptide chains, two heavy chains (H) and two light chains (L), linked together by disulfide bonds, as well as multimers thereof (eg, IgM). Each heavy chain contains a heavy chain variable region (abbreviated herein as VR HC or VH) and a heavy chain constant region. The term "variable region" of an antibody refers to the variable region of an antibody light chain or the variable region of an antibody heavy chain, alone or in combination. The heavy chain constant region contains three domains, CH1, CH2 and CH3. The CH1 and CH2 domains are connected by a hinge region. Each light chain contains a light chain variable region (abbreviated herein as VR LC or VL) and a light chain constant region. The light chain constant region contains one domain (CL1). The VH and VL regions can be further divided into regions of hypervariability, referred to as complementarity determining regions (CDRs), interspersed with regions that are more conserved, referred to as framework regions (FR). Each VH and VL consists of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In various embodiments, FR antibodies against CD3 (or an antigen-binding portion thereof) may be identical to human germline sequences or naturally or artificially modified. The consensus amino acid sequence can be determined by parallel analysis of two or more CDRs. The CDRs on each chain are held in close proximity by the FR and, together with the CDRs on the other chain, participate in the formation of the antigen-binding region of the antibody.

As used herein, the term "antigen-binding fragment" or "antibody fragment" or "antigen-binding moiety" refers to one or more antibody fragments that retain the ability to specifically bind an antigen. Examples of binding fragments included in the term "antigen binding fragment" of an antibody include (i) an antibody heavy chain variable domain (VH), and/or an antibody light chain variable domain (VL), or a VH/VL pair derived from full-length antibodies or fragments antibodies such as VH domain and/or VL domain; (ii) Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH1 domains; (iii) a Fab' fragment that is essentially a Fab with part of the hinge region (eg, Fundamental Immunology, Paul ed., 3.sup.rd ed.1993); (iv) F(ab') ₂ fragment, a bivalent fragment containing two Fab fragments connected by a disulfide bridge in the hinge region; (v) Fd fragment consisting of VH and CH1 domains; (vi) Fv fragment consisting of the VL and VH domains of one arm of the antibody; (vii) single-chain F fragment (scFv), a single protein chain in which the VL and VH regions are paired to form monovalent molecules (eg, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) ) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) disulfide-stabilized Fv moiety (dsFv), an Fv with an engineered intermolecular disulfide bond to stabilize the VH-VL pair; (ix) a single variable domain antibody (sdAb or dAb) (eg, Ward et al., (1989) Nature 341:544-546) consisting of a heavy chain variable domain and lacking a light chain; (x) complementarity determining region (CDR) and any derivatives thereof.

Within the scope of the invention, an "antigen binding fragment" of an antibody may comprise at least one variable domain. The variable domain can be of any size or amino acid composition and will typically contain at least one CDR adjacent or in frame with one or more framework sequences. Within the scope of the invention, an "antigen-binding antibody fragment" may comprise a homodimer or heterodimer (or other multimer) of any of the variable region and constant domain configurations set forth below in non-covalent association with each other and/or with one or more monomeric VH or VL regions (eg , via disulfide bonds). For example, the variable region may be dimeric and contain VH-VH, VH-VL or VL-VL dimers. Variable and constant domain configurations that can be found in the antigen binding fragment of the antibody of the present invention include: VH-CH1; VH-CH2; VH-CH3; VH-CH1-CH2; VH-CH1-CH2-CH3; VH-CH2-CH3; VH-VL-CL, VH-VL-CH1, VH-VL-CH2; VH-CL; VL-CH1; VL-CH2; VL-CH3; VL-CH1-CH2; VL-CH1-CH2-CH3; VL-CH2-CH3 and VL-CL. Variable regions and constant domains can be linked to each other directly or via a full or partial hinge or linker region. The hinge region may consist of at least 2 (eg, 5, 10, 15, 20, 40, 60 or more) amino acids, resulting in flexible or semi-flexible binding between adjacent variable regions and/or constant domains in a single polypeptide molecule.

As used herein, a “binding domain” comprises any region of a polypeptide (eg, an antibody) that is responsible for selective binding to a molecule of interest (eg, an antigen, ligand, receptor, substrate or inhibitor). Examples of binding domains include an antibody variable region, a receptor binding domain, a ligand binding domain, and an enzymatic domain.

As used herein, the term “human acceptor framework” is a framework comprising the amino acid sequence of a variable light chain (VL) framework or a variable heavy chain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. A human acceptor framework "derived from" a human immunoglobulin framework or a human consensus framework may contain the same amino acid sequence or may contain modifications of the amino acid sequence. In some embodiments, the number of amino acid modifications is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the human VL acceptor framework is sequence identical to a human immunoglobulin VL framework sequence or a human consensus framework sequence.

As used herein, the term "affinity mature" antibody refers to an antibody with one or more modifications in one or more variable regions, including CDRs and FRs, compared to a parent antibody not having such modifications, and where such modifications result in an improvement in the antibody's affinity for antigen.

Within the scope of the invention, the term "Fc region", "Fc domain", "Fc chain" or similar terms are used to define the C-terminal region of the IgG heavy chain. The Fc region of IgG contains two constant domains, CH2 and CH3. The CH2 domain of the human IgG Fc region is generally located from amino acid 231 to amino acid 340 according to the EU index numbering system or from amino acid 244 to amino acid 360 according to the Kabat numbering system. The CH3 domain of the human IgG Fc region is generally located from amino acid 341 to amino acid 447 according to the EU index numbering system or from amino acid 361 to amino acid 478 according to the Kabat numbering system. The CH2 domain of the human IgG Fc region (also referred to as the "Cγ 2" domain) is unique in that it is not closely paired with another domain. Instead, two N-linked branched carbohydrate chains are located between the two CH2 domains of intact native IgG. The Fc region may contain native Fc sequences or Fc sequence variants. Although the boundaries of the immunoglobulin heavy chain Fc sequence may vary, the human IgG heavy chain Fc sequence is generally defined as ranging from the amino acid residue at approximately Cys226 or approximately Pro230 to the carboxy terminus of the Fc sequence. Unless otherwise indicated herein, numbering of amino acid residues in the Fc region or constant region is carried out in accordance with the EU numbering system, also referred to as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

In some embodiments, the Fc chain begins at the hinge region immediately upstream of the papain cleavage site and ends at the C terminus of the antibody. Thus, the complete Fc chain contains at least a hinge domain, a CH2 domain, and a CH3 domain. In some embodiments, the Fc chain comprises at least one of: a hinge domain (eg, an upper, middle, and/or lower hinge region), a CH2 domain, a CH3 domain, a CH4 domain, or a variant, portion, or fragment thereof. In some embodiments, the Fc domain comprises a complete Fc chain (ie, a hinge domain, a CH2 domain, and a CH3 domain). In some embodiments, the Fc chain comprises a hinge domain (or a portion thereof) fused to a CH3 domain (or a portion thereof). In some embodiments, the Fc chain comprises a CH2 domain (or a portion thereof) fused to a CH3 domain (or a portion thereof). In some embodiments, the Fc chain consists of or a portion of a CH3 domain. In some embodiments, the Fc chain consists of a hinge domain (or a portion thereof) and a CH3 domain (or a portion thereof). In some embodiments, the Fc chain consists of a CH2 domain (or a portion thereof) and a CH3 domain. In some embodiments, the Fc chain consists of a hinge domain (or a portion thereof) and a CH2 domain (or a portion thereof). In some embodiments, the Fc chain lacks at least a portion of the CH2 domain (eg, all or part of the CH2 domain). The term "Fc chain" generally refers to a polypeptide containing all or part of the Fc chain of an immunoglobulin heavy chain. This includes, by way of non-limiting examples, polypeptides containing the entire CH1 domain, hinge domain, CH2 and/or CH3 domains, as well as fragments of such peptides containing only, for example, the hinge domain, CH2 domain and CH3 domain. The Fc chain can be derived from an immunoglobulin of any species and/or subtype, including, but not limited to, a human IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibody. The Fc domain includes native Fc molecules and variant Fc molecules. As with Fc variants and native Fcs, the term “Fc chain” includes molecules in monomeric or multimeric form, whether cleaved from the whole antibody or produced by other means. In some embodiments, the Fc chain contains the carboxy-terminal portions of both heavy chains held together by disulfide bonds. In some embodiments, the Fc chain consists of a CH2 domain and a CH3 domain.

In this area, the terms "Fc receptor" and "FcR" are used to describe a receptor that binds to the Fc region of an antibody. A preferred FcR is a native human FcR sequence. In addition, a preferred FcR is an IgG antibody-binding FcR (gamma receptor) and includes receptors of the FcγRI, FcγRII and FcγRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences, differing mainly in their cytoplasmic domains. For a review of FcR, see Ravetch and Kinet, Ann. Rev. Immunol., 9:457-92, 1991; Capel et al., Immunomethods, 4:25-34, 1994; and de Haas et al., J. Lab. Clin. Med., 126:330-41, 1995. The term “FcR” also includes the neonatal FcRn receptor, which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol., 117:587, 1976; and Kim et al. , J. Immunol., 24:249, 1994).

A “native Fc region sequence” or “wild-type Fc region” contains an amino acid sequence identical to the amino acid sequence of the Fc region found in nature. The term "wild type" human IgG Fc refers to the amino acid sequence that occurs naturally in the human population. Of course, just as the Fc sequence can vary slightly among individuals, one or more changes can be made to the wild type sequence and it will still be within the scope of the invention. For example, the Fc region may contain additional changes not relevant to the present invention, such as a mutation in a glycosylation site, the inclusion of an unnatural amino acid, or a knob-to-groove mutation.

An "Fc region variant" or "Fc chain variant" contains an amino acid sequence that differs from the native Fc region sequence by at least one amino acid modification, but retains at least one effector function of the native Fc region sequence. In some embodiments, the Fc chain variant has at least one amino acid substitution relative to the native Fc chain or Fc region sequence of the parent polypeptide, for example, from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in the native sequence of the Fc chain or the Fc chain of the parent polypeptide. As used herein, the Fc chain variant will preferably have at least about 80% sequence identity to the native Fc chain sequence and/or the Fc chain of the parent polypeptide, and most preferably at least about 90% sequence identity, more preferably, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% sequence identity.

As used herein, the term “effector functions” refers to the biological activities attributable to the Fc chain (either the native Fc chain sequence or the amino acid sequence of a variant Fc chain) of an antibody and varies depending on the isotype of the antibody. Examples of antibody effector functions include: C1q binding and complement-mediated cytotoxicity; Fc receptor binding; antibody-dependent cellular cytotoxicity (ADCC); phagocytosis; negative regulation of cell surface receptors (eg, B cell receptor) and B cell activation. Such effector functions typically require the combination of an Fc chain with a binding domain (eg, an antibody variable region) and can be assessed using various assays known in the art for assessing such antibody effector functions. An example of measuring effector function is by Fcγ3 and/or C1q binding.

The effector functions of antibodies are determined by the sequences of the Fc chain; this chain is also the part recognized by Fc receptors (FcRs) found on some cell types.

In some embodiments, the Fc polypeptide comprises part or all of a wild-type hinge region sequence (typically at the N terminus). In some embodiments, the Fc polypeptide does not contain a functional hinge sequence or a wild-type hinge sequence.

As used herein, the term “hinge region,” “hinge sequence,” and variations thereof include the meaning known in the art, illustrated, for example, in Janeway et al., ImmunoBiology: the immune system in health and disease, Elsevier Science Ltd., NY (4th ed., 1999); Bloom et al., Protein Science, 6:407-415, 1997; and Humphreys et al., J. Immunol. Methods, 209:193-202, 1997.

As used herein, the term "immunoglobulin-like hinge region", "immunoglobulin-like hinge sequence" or variants thereof refers to the hinge region and hinge sequence of an immunoglobulin-like or antibody-like molecule (eg, immunoadhesins). In some embodiments, the immunoglobulin-like hinge region can be derived from any IgG1, IgG2, IgG3, or IgG4 subtype, or from IgA, IgE, IgD, or IgM, including chimeric forms thereof, such as an IgG1/2 chimeric hinge region.

In some embodiments, the hinge region can be derived from the human IgG1 subtype from amino acid 216 to amino acid 230 according to the EU index numbering system, or from amino acid 226 to amino acid 243 according to the Kabat numbering system. In some embodiments, the sequence may be EPKSCDKTHTCPPCP (SEQ ID NO: 186). Those skilled in the art may differ in their understanding of the specific amino acids corresponding to the various domains of the IgG molecule. Thus, the N-terminus or C-terminus of the domains described above may be longer or shorter by 1, 2, 3, 4, 5, 6, 7, 8, 9 or even 10 amino acids.

In some embodiments, the hinge region may be mutated by one or more amino acids. In some embodiments, the hinge region may be shortened and comprise only a portion of the full hinge region. In some embodiments, the hinge region may comprise only the last 5 amino acids of the hinge region.

Within the scope of the invention, the terms "linked", "fused", "fusion", "covalently linked", "covalently linked" and "genetically fused" are used interchangeably. These terms refer to the joining of two or more elements or components by any means, including chemical conjugation or recombinant methods. As used herein, the term “covalently linked” means that the moieties are directly covalently linked to each other or indirectly covalently linked to each other by an intermediate residue or residues, such as a linker peptide or fragment. In a preferred embodiment, the fragments are covalently fused. One type of covalent bond is a peptide bond. Chemical conjugation methods (eg, using heterobifunctional cross-linkers) are known in the art. The fused fragments can also be subjected to genetic fusion. As used herein, the term “genetically fused” or “genetically linked” refers to the co-linear covalent bonding or joining of two or more proteins, polypeptides or fragments thereof through their separate peptide backbones, through the genetic expression of a single polynucleotide molecule encoding those proteins, polypeptides or fragments. This genetic fusion results in the expression of a single contiguous genetic sequence. The preferred genetic fusion is carried out in frame, i.e. two or more open reading frames (ORFs) are fused to form a contiguous longer ORF in a manner that maintains the correct reading frame of the original ORFs. Thus, the resulting recombinant fusion protein is a single polypeptide containing two or more protein segments corresponding to the polypeptides encoded by the original ORFs (the segments are not normally connected in this way in nature). In this case, a single polypeptide is cleaved during processing to form dimeric molecules containing two polypeptide chains.

As used herein, the term “modification” refers to the substitution, insertion and/or deletion of an amino acid in a polypeptide sequence, a change in a moiety chemically linked to a protein, or a modification of the function of a protein, such as an antibody. For example, the modification may be an altered function of the antibody or an altered carbohydrate structure attached to the protein. As used herein, the term "amino acid modification" refers to a mutation, insertion, or deletion of one or more amino acid residues in an antibody. The term "amino acid mutation" means the replacement of at least one existing amino acid residue with another different amino acid residue (eg, amino acid residue substitution). The term "amino acid deletion" means the removal of at least one amino acid residue at a predetermined position in the amino acid sequence. For example, the L234A mutation means that the amino acid residue lysine at position 234 in the Fc region of the antibody is replaced by the amino acid residue alanine (lysine alanine substitution) (numbering according to the EU index). The term "naturally occurring amino acid residue" means an amino acid residue from the group consisting of alanine (three-letter symbol: Ala, single-letter symbol: A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine (Cys, C), glutamine (Gln, Q), glutamic acid (Glu, E), glycine (Gly, G), histidine (His, H), isoleucine (Ile, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine ( Tyr, Y) and valine (Val, V).

As used herein, a “conservative amino acid substitution” is a substitution in which an amino acid residue is replaced with an amino acid residue having a similar side chain. In this region, families of amino acid residues that have similar side chains are identified. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., asparagine, glutamine, serine, threonine, tyrosine, cysteine) , nonpolar side chains (e.g., glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine , tryptophan, histidine).

For purposes of the invention, a "nonessential" amino acid residue is one that can be changed relative to the wild-type sequence of a binding agent, eg an antibody, without eliminating or significantly altering the biological activity, whereas a change in an "essential" amino acid residue results in such a change. In an antibody, an essential amino acid residue may be a specificity determining residue (SDR).

The term "agent" is used herein to refer to a biological macromolecule, an extract from biological materials, a mixture of biological macromolecules, a chemical compound, a mixture of chemical compounds, and/or a mixture of chemical compounds and biological macromolecules. The term "therapeutic agent" refers to an agent having biological activity.

As used herein, the term "monoclonal antibody" refers to an antibody derived from a population of substantially homogeneous antibodies, i.e. The individual antibodies that make up the population are identical, except for possible natural mutations that may be present in minute quantities. Monoclonal antibodies are highly specific and directed against a single antigenic site. Additionally, unlike polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against the same determinant on an antigen. The designation "monoclonal" indicates the nature of the antibody as being derived from an essentially homogeneous population of antibodies, and should not be construed as requiring the antibody to be produced by any particular method. For example, monoclonal antibodies for use in the present invention can be produced by the hybridoma method first described in Kohler and Milstein, Nature 256:495, 1975, or by recombinant DNA methods as described in US Pat. No. 4,816,567. Monoclonal antibodies can also be isolated from phage libraries , obtained by the method described, for example, in McCafferty et al., Nature 348:552-554, 1990.

Antibodies of the present invention may be "humanized antibodies". As used herein, the term “humanized” antibody refers to non-human forms of antibodies (e.g., mouse, rat, rabbit, non-human primate, or other mammal) that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab ', F(ab') ₂ or other antigen-binding antibody subsequences) containing one or more amino acid residues incorporated therein from a non-human source. These non-human amino acid residues are often referred to as “imported” residues, typically derived from an “imported” variable domain. The imported residue, sequence or antibody has the desired affinity and/or specificity or other desired biological activity of the antibody as described herein.

Preferably, the humanized antibodies are human immunoglobulins (recipient antibody) in which residues from the complementarity determining region (CDR) of the recipient are replaced with CDR residues from a non-human species (donor antibody), such as mouse, rat or rabbit, having the desired specificity, affinity and capacity . In some cases, human immunoglobulin Fv framework region (FR) residues are replaced with corresponding non-human residues. In addition, the humanized antibody may contain residues not found in either the recipient antibody or the imported CDR or framework sequences, but included to further improve and optimize the properties of the antibodies. In general, a humanized antibody will contain substantially all of at least one and typically two variable regions, in which all or substantially all of the CDR regions correspond to the CDR of a non-human immunoglobulin, and all or, at Essentially, all of the FR regions are regions from the human immunoglobulin consensus sequence. The humanized antibody will optimally also comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically from a human immunoglobulin. Preferred are antibodies having Fc chains modified as described in WO 99/58572. Other forms of humanized antibodies have one or more CDRs (CDR L1, CDR L2, CDR L3, CDR H1, CDR H2, or CDR H3) modified from the parent antibody, also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody. As used herein, the term “humanized” is intended to include deimmunized antibodies.

As used herein, the term “human antibody” means an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and/or produced by any of the methods for producing human antibodies known to those skilled in the art or described herein. Thus, for purposes of the invention, the term “human antibody” is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (eg, mutations introduced by random or site-specific mutagenesis in vitro or somatic mutation in vivo ), for example, in the CDR, and in particular, CDR3. This definition of human antibody includes antibodies containing at least one human heavy chain polypeptide or at least one human light chain polypeptide. One such example is an antibody containing mouse light chain and human heavy chain polypeptides. Human antibodies can be obtained by various methods known in this field. In one embodiment, the human antibody is selected from a phage library, wherein the phage library expresses human antibodies (Vaughan et al., Nature Biotechnology, 14:309-314, 1996; Sheets et al., Proc. Natl. Acad. Sci. (USA ) 95:6157-6162, 1998; Hoogenboom and Winter, J. Mol. Biol., 227:381, 1991; Marks et al., J. Mol. Biol., 222:581, 1991). Human antibodies can also be obtained by immunizing animals into which human immunoglobulin loci have been transgenically inserted in place of endogenous loci, for example mice in which endogenous immunoglobulin genes are partially or completely inactivated. This approach is described in US Pat. Nos. 5,545,807, 5,545,806, 5,569,825, 5,625,126, 5,633,425, and 5,661,016. Alternatively, a human antibody can be produced by immortalizing human B lymphocytes that produce antibody against a target antigen (such B lymphocytes can be isolated from an individual or cloning individual cells using cDNA, or they can be immunized in vitro ). See, for example, Cole et al. Monoclonal antibodies and Cancer Therapy, Alan R. Liss, p. 77, 1985; Boerner et al., J. Immunol., 147(1):86-95, 1991; and US Patent No. 5750373.

The human antibodies of the present invention can exist in at least two forms due to the heterogeneity of the hinge region. For example, the immunoglobulin molecule contains a stable four-chain construct of approximately 150-160 kDa in which the dimers are held together by interchain disulfide bonds between the heavy chains. Alternatively, the dimers are not linked by interchain disulfide bonds, and a molecule of approximately 75-80 kDa is formed, consisting of covalently linked light and heavy chains (half-antibody).

For the purposes of the invention, the term "recombinant human antibody" is intended to include all human antibodies produced, expressed, generated or isolated by recombinant methods, such as antibodies expressed using a recombinant expression vector with which a host cell is transfected (described below), antibodies , isolated from a recombinant combinatorial library of human antibodies (described below), antibodies isolated from an animal (for example, a mouse) that is transgenic for human immunoglobulin genes (see, for example, Taylor et al. (1992) Nucl. Acids Res. 20: 6287-6295), or antibodies produced, expressed, created or isolated by any other means involving splicing human immunoglobulin gene sequences with other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, if an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis), and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, although derived from and related to human germline VH and VL sequences, may not naturally exist in the human germline antibody repertoire in vivo .

Antibodies of the invention may be "chimeric" antibodies in which a portion of the heavy and/or light chain is identical or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular class or subclass of antibodies, while the remainder of the chains are identical or homologous to corresponding sequences in antibodies derived from other species or belonging to another class or subclass of antibodies, as well as fragments of such antibodies, provided that they exhibit the desired biological activity (US patent No. 4816567; and Morrison et al., Proc. Natl. Acad Sci USA 81:6851-6855, 1984). Chimeric antibodies of interest herein include primatized antibodies containing antigen binding variable region sequences derived from a non-human primate (eg, Old World monkeys, great apes, etc.) and human constant region sequences.

A "monovalent antibody" contains one antigen-binding site per molecule (eg, IgG or Fab). In some cases, a monovalent antibody may have more than one antigen-binding site, but the binding sites are for different antigens.

The term "monospecific antibody" refers to an antibody or antibody preparation containing two identical antigen-binding sites per molecule (eg, IgG) such that the two binding sites bind to an identical epitope on the antigen. Thus, they compete with each other for binding to the same antigen molecule. This term includes "monoclonal antibody" or "monoclonal antibody composition". Most antibodies found in nature are monospecific. In some cases, the monospecific antibody may be a monovalent antibody (eg, Fab).

A "bivalent antibody" contains two antigen-binding sites per molecule (eg, IgG). In some cases, the two binding sites have the same antigen specificity. However, bivalent antibodies can be bispecific.

As used herein, a "bispecific antibody", "dual specific antibody", "bifunctional antibody", "heteromultimer", "heteromultimeric complex", "bispecific heterodimeric diabody" or "heteromultimeric polypeptide" is a molecule comprising at least a first polypeptide and a second polypeptide, wherein the second polypeptide differs in amino acid sequence from the first polypeptide by at least one amino acid residue. In some cases, a bispecific antibody is an artificial hybrid antibody having two different heavy chain regions and a light chain region. Preferably, the bispecific antibody has binding specificity for at least two different ligands, antigens or binding sites. Thus, bispecific antibodies can bind simultaneously to two different antigens. The two antigen-binding regions of a bispecific antibody bind to two different epitopes, which may be on the same or different protein targets, such as a tumor target.

As used herein, the term "target antigen", "target cell antigen", "tumor antigen" or "tumor-specific antigen" refers to an antigenic determinant present on the surface of a target cell, for example a cell in a tumor, such as a malignant cell or cell tumor stroma.

The terms "mutation load" are used interchangeably herein. Tumor mutational load is a measure of the number of mutations in the tumor genome, defined as the total number of mutations per coding region of the tumor genome. Considerable variability in mutational load is observed among tumor types, ranging from just a few mutations to thousands of mutations (Alexandrov LB et al., Nature 2013;500(7463):415-421; Lawrence MS et al., Nature 2013;499:214- 218; Vogelstein B et al., Science, 2013;339:1546-1558).

Within the scope of the invention, a first polypeptide chain and a second polypeptide chain of a "bispecific antibody" comprise at least one antibody VL and one antibody VH or fragment thereof, wherein both antibody binding domains are contained in a single polypeptide chain, and wherein the VL and VH regions in each polypeptide chain derived from different antibodies.

A bispecific antibody, a dual-specific antibody, a bifunctional antibody, a heteromultimer, a heteromultimeric complex, a bispecific heterodimeric diabody, or a heteromultimeric polypeptide can form higher order tertiary structures where other polypeptides are present in addition to the first and second polypeptide. The heteromultimer polypeptides can interact with each other through non-peptide covalent bonds (eg, disulfide bonds) and/or non-covalent interactions (eg, hydrogen bonds, ionic bonds, van der Waals forces and/or hydrophobic interactions).

A bispecific antibody, dual-specific antibody, bifunctional antibody, heteromultimer, heteromultimeric complex, bispecific heterodimeric diabody, or heteromultimeric polypeptide can be prepared by designing sFv fragments with short linkers (eg, approximately 3-10 residues) between the VH and VL regions such that achieve interchain rather than intrachain pairing of V regions, which leads to the formation of a bivalent fragment, i.e. fragment having two antigen-binding sites. Bispecific antibodies can be prepared from full-length antibodies or antibody fragments (eg, F(ab′) ₂ bispecific antibodies). Diabodies are described in more detail, for example, in EP404,097; WO93/11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448, 1993. Bispecific antibodies are heterodimers of two “crossover” sFv fragments, in which the VH and VL regions of the two antibodies are on different polypeptide chains.

As used herein, the term "isolated antibody" means an antibody that has been identified and separated and/or isolated from at least one component from its natural environment. For example, an antibody isolated or removed from at least one component of the body or from a tissue or cell in which the antibody exists or is naturally produced is an “isolated antibody” for purposes of the present invention. The isolated antibody also includes the antibody in situ in the recombinant cell. Isolated antibodies are antibodies that have been subjected to at least one purification or isolation step. In some embodiments, the isolated antibody may be substantially free of other cellular material and/or chemicals.

As used herein, the term “linked” or “linkage” refers to a direct connection via a peptide bond between a first and second amino acid sequence, or a connection comprising a third amino acid sequence linked by a peptide bond to the first and second amino acid sequences. For example, a linker connected by a peptide bond to the C-terminus of one amino acid sequence and the N-terminus of another amino acid sequence.

As used herein, the term “linker” refers to an amino acid sequence of two or more amino acids in length. The linker may consist of neutral polar or non-polar amino acids. The linker may be, for example, 2 to 100 amino acids in length, for example, 2 to 50 amino acids in length, for example, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 amino acids in length . The linker may be "cleavable", for example, by self-cleavage or enzymatic or chemical cleavage. Cleavage sites in amino acid sequences and enzymes and chemicals that cleave at such sites are well known in the art and are also presented herein.

As used herein, the term "disulfide bond" or "cysteine-cysteine disulfide bond" refers to a covalent interaction between two cysteines in which the sulfur atoms of the cysteines are oxidized to form a disulfide bond. The average binding energy for a disulfide bond is approximately 60 kcal/mol compared to 1-2 kcal/mol for a hydrogen bond. In the context of the present invention, disulfide bond-forming cysteines are located in the framework regions of the single chain antibody and serve to stabilize the conformation of the antibody. Cysteine residues can be inserted, for example, by site-directed mutagenesis in such a way that stabilizing disulfide bonds can be formed in the molecule.

The designation "ridge-to-recess" is similar to the designation "ridge-to-recess" and can be used interchangeably.

The term “protrusion” refers to at least one amino acid side chain protruding from the contact surface of the first polypeptide and thus positioned in a compensatory cavity in the adjacent contact surface (i.e., the contact surface of the second polypeptide) to stabilize the heterodimer, thereby thus promotes the formation of a heterodimer compared to the formation of a homodimer. The protrusion may exist on the original contact surface, or it may be inserted synthetically (eg, by measuring the nucleic acid encoding the contact surface). Typically, the nucleic acid encoding the contact surface of the first polypeptide is modified so that it encodes the protrusion. To achieve this, the nucleic acid encoding at least one original amino acid residue at the contact surface of the first polypeptide is replaced with a nucleic acid encoding at least one imported amino acid residue having a larger side chain volume than the original amino acid residue. The upper limit on the number of original residues that are replaced is the total number of residues in the contact surface of the first polypeptide. Some of the imported residues to form the protrusion are typically natural amino acid residues and are preferably selected from arginine (R), phenylalanine (F), tyrosine (Y) and tryptophan (W).

The protrusion is "located" in the recess, which means that the spatial arrangement of the protrusion and recess on the contact surface of the first polypeptide and the second polypeptide, respectively, and the dimensions of the protrusion and recess are such that the protrusion can be located in the recess without significantly disrupting the normal connection of the first and second polypeptides in the contact surface. Because protrusions such as phenylalanine (F), tyrosine (Y) and tryptophan (W) are generally not perpendicular to the axis of the contact surface, the alignment of a protrusion with the corresponding valley is based on modeling the protrusion/valley pair based on a three-dimensional structure, such as that obtained by X-ray crystallography or nuclear magnetic resonance (NMR). This can be achieved by methods widely used in this field.

The term “groove” refers to at least one amino acid side chain recessed relative to the contact surface of the second polypeptide and thereby receiving a corresponding protrusion on the adjacent contact surface of the first polypeptide. The depression may exist on the original contact surface, or it may be inserted synthetically (eg, by altering the nucleic acid encoding the contact surface). Typically, the nucleic acid encoding the contact surface of the second polypeptide is modified so that it encodes a cavity. To achieve this, the nucleic acid encoding at least one original amino acid residue at the contact surface of the second polypeptide is replaced with DNA encoding at least one “imported” amino acid residue having a smaller side chain volume than the original amino acid residue. The upper limit on the number of original residues that are replaced is the total number of residues at the interface of the second polypeptide. Some of the imported residues for forming the cavity are typically natural amino acid residues and are preferably selected from alanine (A), serine (S), threonine (T) and valine (V).

As used herein, the terms "contact surface", "contact surface residue", "contact surface amino acid", "contact residue" or "contact amino acid" generally refer to any amino acid residue present in a domain that can participate in contacts between a first polypeptide and a second polypeptide.

The "original amino acid" residue is the residue that is replaced by the "imported amino acid" residue, which may have a smaller or larger side chain than the original residue. The imported amino acid residue may be a natural or unnatural amino acid residue, but is preferably the former. "Natural" amino acid residues are residues encoded by the genetic code. The term "unnatural" amino acid residue means a residue that is not encoded by the genetic code, but is capable of covalently binding to adjacent amino acid residues of the polypeptide chain. Examples of unnatural amino acid residues are norleucine, ornithine, norvaline, homoserine and other amino acid residue analogs such as those described, for example, in Ellman et al., Meth. Enzyme. 202:301-336 (1991). In some embodiments, the method of the present invention may include replacing at least one original amino acid residue, but more than one original amino acid residue may be replaced. Typically, no more than the total number of residues at the contact surface of the first or second polypeptide may contain the original amino acid residues being replaced.

As used herein, the term “competes” with respect to an antibody means that the first antibody or antigen-binding portion (or portion) thereof binds to an epitope substantially similar to the binding of the second antibody or antigen-binding portion thereof, such that the result of the first antibody binding to its cognate epitope is detectably reduced in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody. An alternative option, in which the binding of the second antibody to its epitope is also detectably reduced in the presence of the first antibody, may occur, but is not required. Those. the first antibody can inhibit the binding of the second antibody to its epitope without the second antibody inhibiting the binding of the first antibody to its corresponding epitope. However, if each antibody detectably inhibits the binding of another antibody to its cognate epitope or ligand to the same, greater, or lesser extent, the antibodies are said to “cross-compete” with each other for binding to their respective epitopes. Competing and cross-competing antibodies are within the scope of the present invention. Regardless of the mechanism by which such competition or cross-competition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope or portion thereof), those skilled in the art, given the teachings presented herein, will appreciate that such competing and/or cross-competing antibodies, and can be used for the methods presented herein.

As used herein, the term "antibody-dependent cellular cytotoxicity" or "ADCC" refers to a cell-mediated reaction in which non-specific cytotoxic cells expressing Fc receptors (FcR) (eg, natural killer (NK) cells, neutrophils and macrophages) recognize bound antibody on a target cell and then cause lysis of the target cell. The ADCC activity of a molecule of interest can be assessed using an in vitro ADCC assay as described in US Patent Nos. 5,500,362 or 5,821,337. Effector cells that can be used for such assays include peripheral blood mononuclear cells (PBMCs) and NK cells. Alternatively or additionally, the ADCC activity of a molecule of interest can be assessed in vivo , for example, in an animal model as described in Clynes et al., PNAS (USA), 95:652-656, 1998.

As used herein, the term “complement-mediated cytotoxicity” or “CDC” refers to the lysis of a target in the presence of complement. The complement activation pathway is initiated by the binding of the first component of the complement system (C1q) to a molecule (eg, antibody) complexed with a cognate antigen. To assess complement activation, a CDC assay can be performed, for example, as described in Gazzano-Santoro et al., J. Immunol. Methods, 202: 163, 1996.

As used herein, the terms “immunospecifically binds,” “immunospecifically recognizes,” “specifically binds,” “specifically recognizes,” and similar terms refer to molecules, such as binding domains, that specifically bind an antigen (e.g., an epitope or immune complex) and that do not bind specifically with another molecule. A molecule that specifically binds an antigen may bind to other peptides or polypeptides with lower affinity, as determined by assays known in the art, such as immunoassays, BIACORE™, or other assays. Preferably, molecules that specifically bind to the antigen do not cross-react with other proteins.

Suitable "moderate stringency conditions" include pre-wash in a solution of 5x SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridization at 50°C-65°C, 5x SSC, overnight; followed by washing twice at 65°C for 20 minutes with each of 2×, 0.5×, and 0.2× SSC containing 0.1% SDS.

Within the scope of the invention, the “high stringency conditions” are as follows: (1) the wash uses low ionic strength and high temperature, for example, 0.015 M sodium chloride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50° C.; (2) a denaturing agent such as formamide is used during hybridization, e.g. 50% (v/v) formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM buffer sodium phosphate at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42°C; or (3) use 50% formamide, 5x SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5x Denhardt's solution, sonicated salmon milk DNA (50 µg/ml), 0.1% SDS and 10% dextran sulfate at 42°C with washes at 42°C in 0.2x SSC (sodium chloride/sodium citrate) and 50% formamide at 55°C followed by a high stringency wash consisting of 0.1× SSC containing EDTA at 55°C. Those skilled in the art will understand how to adjust temperature, ionic strength, etc. as necessary to adapt to factors such as probe length, etc.

Binding proteins, binding domains, CDRs or antibodies (defined herein broadly) can be identified according to the Kabat, Chothia, Kabat and Chothia, AbM, Contact, North and/or conformational definitions, or any CDR definition method. well known in this field. See, for example, Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th ed. (hypervariable regions); Chothia et al., Nature 342:877-883, 1989 (structural loop structures). The identity of the amino acid residues in a particular antibody that make up the CDR can be determined by methods well known in the art. The AbM CDR determination is a compromise between Kabat and Chothia and uses Oxford Molecular's AbM antibody modeling software (Accelrys®). The "Contact" CDR determination is based on observed antigen contacts as described in MacCallum et al., J. Mol. Biol., 262:732-745, 1996. The “conformational” definition of CDR is based on residues that make an enthalpic contribution to antigen binding (see, for example, Makabe et al., J. Biol. Chem., 283:1156-1166, 2008). North identified canonical CDR conformations using another preferred set of CDR definitions (North et al., J. Mol. Biol. 406: 228-256, 2011). In another approach, referred to herein as “conformational sensing” of CDRs, CDR positions can be identified as residues that contribute enthalpicly to antigen binding (Makabe et al., J Biol. Chem. 283:1156-1166, 2008). In other boundary definitions, the CDRs may not strictly follow one of the approaches described above, but will nonetheless overlap with at least part of the Kabat CDRs, although they may be shorter or longer in light of prediction or experimental results regarding whether that specific residues, or groups of residues, or even entire CDRs, do not significantly affect antigen binding. For purposes of the invention, the term "CDR" may refer to CDRs determined using any approach known in the art, including combinations of approaches. The methods used herein may use CDRs determined using any of these approaches. In the case of any of these embodiments containing more than one CDR, the CDR (or other antibody residue) can be defined in accordance with any of the definitions of Kabat, Chothia, extended definition (combinations of Kabat and Chothia), North, extended definition, AbM, Contact and /or conformational determinations.

Residues in the variable domain are numbered Kabat, which is the numbering system used for heavy chain variable regions or light chain variable regions of antibody compilation. See Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991. Using this numbering system, a particular linear amino acid sequence may contain fewer amino acids or additional amino acids corresponding to a truncation or insertion in the FR or CDR of the variable region. For example, a heavy chain variable region may include a single amino acid insertion (Kabat residue 52a) after H2 residue 52 and inserted residues (e.g., Kabat residues 82a, 82b, and 82c) after heavy chain FR residue 82. Kabat residue numbering can be determined for the specified antibody by aligning regions of homology of the antibody sequence with the “standard” Kabat numbered sequence. Various algorithms are available for Kabat numbering. The algorithm embodied in version 2.3.3 of the Abysis release (www.abysis.org) was used in the present invention to assign Kabat numbering to the variable regions CDR1 VL, CDR2 VL, CDR3 VL, CDR2 VH and CDR3 VH. AbM determination was used for CDR1 VH. Specific positions of amino acid residues in an antibody can also be numbered according to Kabat.

As used herein, the term “phage display library” refers to a population of bacteriophages, each of which contains foreign cDNA recombinantly fused in frame to a surface protein. The phage displays the foreign protein encoded by the cDNA on its surface. After replication in a bacterial host cell, typically E. coli , a phage containing the foreign cDNA of interest is selected for expression of the foreign protein on the phage surface.

The term "epitope" refers to a portion of a molecule that can be recognized, contacted and/or bound by an antibody at one or more antigen-binding regions of the antibody, known as a paratope. One antigen can have more than one epitope. Thus, different antibodies may bind to different regions on the antigen and may have different biological effects. Epitopes often consist of a chemically active surface grouping of molecules, such as amino acids or sugar side chains, and have specific three-dimensional structural characteristics as well as specific charge characteristics. Within the scope of the invention, epitopes can be conformational or linear. A conformational epitope is formed by spatially close amino acids from different segments of a linear polypeptide chain. A linear epitope is an epitope formed by adjacent amino acid residues in a polypeptide chain. In some embodiments, the epitope may include moieties of saccharides, phosphoryl groups, or sulfonyl groups on the antigen.

As used herein, the term "antigenic epitope" is defined as that portion of a polypeptide to which an antibody can specifically bind, as determined by any method well known in the art, for example, conventional immunoassays. A "non-linear epitope" or "conformational epitope" contains non-contiguous polypeptides (or amino acids) in an antigenic protein to which an epitope-specific antibody binds. Once the desired epitope on an antigen has been determined, antibodies to that epitope can be produced, for example, by the methods presented herein.

The terms "specific binding" or "specifically binding" when used in relation to the interaction of an antibody and a protein or peptide refer to an interaction dependent on the presence of a particular structure (ie, antigenic determinant or epitope) on the protein; in other words, the antibody recognizes and binds to a specific protein structure rather than to proteins in general. For example, if an antibody is specific for epitope "A", the presence of protein containing epitope A (or free, unlabeled A) in a reaction mixture containing labeled "A" and antibody will reduce the amount of labeled A bound to the antibody.

In some embodiments, the term “specifically binds” means, for example, that the antibody binds to a protein with a K _D of about 0.1 nM or less, but more typically less than about 1 μM. In some embodiments, the term “specifically binds” means that the antibody binds to the target, sometimes with a K _D of at least about 0.1 μM or less, and in other cases with at least about 0.01 μM or less, and in in other cases, at least about 1 nM or less. Because of the sequence identity between homologous proteins in different species, specific binding may involve an antibody that recognizes a protein in more than one species (eg, human GUCY2c and mouse GUCY2c). Likewise, because of homology in some regions of the polypeptide sequences of different proteins, specific binding may involve an antibody recognizing more than one protein. It should be understood that in some embodiments, an antibody or binding fragment that specifically binds to a first target may or may not bind specifically to a second target. In this regard, "specific binding" does not necessarily require (although it may include) exclusive binding, i.e. binding to a single target. Thus, in some embodiments, an antibody may specifically bind to more than one target. In some embodiments, multiple targets may be bound by the same antigen-binding site on an antibody. For example, in some cases, an antibody may contain two identical antigen-binding sites, each of which specifically binds to the same epitope on two or more proteins. In some alternative embodiments, the antibody may be multispecific and contain at least two antigen binding sites with different specificities. By way of non-limiting example, a bispecific antibody may comprise one antigen binding region that recognizes an epitope on one protein (eg, human CD3) and further comprise a second, different antigen binding region that recognizes a different epitope on a second protein. Typically, but not necessarily, a reference to binding means a specific binding.

An antibody that specifically binds to an antigen may bind to other peptides or polypeptides with lower affinity, as determined by assays known in the art, such as immunoassays, BIACORE ™ or other assays. Preferably, the antibody that specifically binds to the antigen does not cross-react with other proteins.

The terms "nonspecific binding" or "background binding" when used in relation to an antibody-protein or peptide interaction refer to an interaction that is independent of the presence of a specific structure (i.e., the antibody binds to proteins in general rather than to a specific structure such as epitope).

As used herein, the term "k _on " or "k _a " refers to the rate constant of association of an antibody with an antigen. In particular, rate constants (k _on /k _a and k _off /k _d ) and equilibrium dissociation constants are measured using whole antibody (ie, bivalent) and monomeric GUCY2c proteins.

As used herein, the term "k _off " or "k _d " refers to the rate constant for the dissociation of an antibody from an antibody/antigen complex.

As used herein, the term "K _D " refers to the equilibrium dissociation constant of an antibody-antigen interaction.

As used herein, the term “binding affinity” generally refers to the strength of the sum of all non-covalent interactions between one binding site of a molecule (eg, an antibody) and its binding partner (eg, an antigen). Unless otherwise specified, as used herein, the term “binding affinity” refers to the characteristic binding affinity reflecting the 1:1 interaction between members of a binding pair (eg, antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by a dissociation constant ( _KD ). The affinity of a molecule X for its partner Y can generally be represented by a dissociation constant (Kd). For example, Kd may be approximately 200 nM, 150 nM, 100 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 8 nM, 6 nM, 4 nM, 2 nM, 1 nM or less. Affinity can be measured by conventional methods known in the art, including the methods presented herein. Low-affinity antibodies tend to bind to antigen slowly and tend to dissociate quickly, while high-affinity antibodies tend to bind to antigen more quickly and tend to remain bound for a longer period of time. Various methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present invention. In particular, the term "binding affinity" is intended to refer to the rate of dissociation of a particular antigen-antibody interaction. K _D is the ratio of the rate of dissociation, also called the "reverse reaction rate (k _off )", and the rate of association or "forward reaction rate (k _on )". Thus, K _D is equal to k _off /k _on and is expressed as molar concentration (M). It follows that the lower the K _D , the stronger the binding affinity. Thus, K _D 1 μM indicates weak binding affinity compared to K _D 1 nM. Antibody K _D values can be determined by methods well known in the art. One way to determine the K _D of an antibody is by using surface plasmon resonance (SPR), typically using a biosensor system such as the BIACORE™ system. The BIACORE™ kinetic assay involves analyzing the binding and dissociation of antigen from chips with immobilized molecules (eg, molecules containing epitope-binding domains) on their surface. Another way to determine the K _D of an antibody is to use biolayer interferometry, typically using OCTET ^® technology (Octet QK ^e system, ForteBio).

The terms "biologically active", "biological activity" and "biological characteristics" in relation to a bispecific antibody of the present invention, such as an antibody, fragment or derivative thereof, mean having the ability to bind to a biological molecule unless otherwise specified.

As used herein, the terms "nucleic acids" and "nucleotide sequences" include DNA molecules (eg, cDNA or genomic DNA), RNA molecules (eg, mRNA), combinations of DNA and RNA molecules or hybrid DNA/RNA molecules, and analogues of DNA or RNA molecules . Such analogs can be prepared using, for example, nucleotide analogs including, but not limited to, inosine or tritylated bases. Such analogues may also contain DNA or RNA molecules containing modified backbones that impart beneficial properties to the molecules, such as, for example, nuclease resistance or increased ability to penetrate cell membranes. The nucleic acids or nucleotide sequences may be single-stranded, double-stranded, may contain single-stranded and double-stranded portions, and may contain triple-stranded portions, but is preferably double-stranded DNA.

The present invention also relates to polynucleotides encoding the antibodies of the invention, including polypeptides and antibody binding regions. Polynucleotides encoding the molecules of the invention can be prepared and the nucleotide sequence of the polynucleotides determined by any method known in the art.

Polynucleotides encoding antibodies of the present invention may include the following: only the coding sequence for the variant, the coding sequence for the variant, and additional coding sequences such as a functional polypeptide, or a signal or secretory sequence or a protein sequence; a coding sequence for the antibody; and a non-coding sequence, such as introns or 5' and/or 3' non-coding sequence of the antibody coding sequence. The term "antibody-encoding polynucleotide" includes a polynucleotide including additional coding sequence for a variant, but also a polynucleotide including additional coding and/or non-coding sequence. It is known in the art that a polynucleotide sequence optimized for a particular host cell/expression system can be easily derived from the amino acid sequence of the desired protein (see ^GENEART® AG, Regensburg, Germany).

The antibodies and antigen binding fragments thereof of the present invention may have additional conservative amino acid substitutions or nonessential amino acid substitutions that do not have a significant effect on the functions of the polypeptide. Whether a particular replacement will be well tolerated or not, i.e. whether it will negatively affect desired biological properties, such as binding activity, can be determined as described in Bowie, JU et al. Science 247: 1306-1310,1990 or Padlan et al. FASEB J. 9: 133-139, 1995.

As used herein, the term "isolated" refers to material removed from its original environment (eg, the natural environment, if natural). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide separated from some or all of the accompanying materials in a natural system is isolated. Such a polynucleotide may be part of a vector, and/or such a polynucleotide or polypeptide may be part of a composition, such as a mixture, solution or suspension, containing an isolated cell or cultured cell containing the polynucleotide or polypeptide, and will still be isolated in the sense that the vector or the composition is not part of its natural environment.

As used herein, the term "replicon" refers to any genetic element, such as a plasmid, chromosome or virus, that functions as an autonomous polynucleotide replication unit in a cell.

Within the scope of the invention, the term "operably linked" refers to a situation in which the described components are in a relationship that allows them to function in a predetermined manner. For example, a control sequence "operably linked" to a coding sequence is ligated so as to achieve expression of the coding sequence under conditions appropriate or compatible with the control sequence. In general, the term "operably linked" means that the linked DNA sequences are contiguous and, in the case of a secretory leader sequence, contiguous and in frame. However, enhancers may not be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such regions are not available, synthetic oligonucleotide adapters or linkers are used in accordance with standard practice.

As used herein, the term “vector” means a construct that can deliver and preferably express one or more genes or sequences of interest in a host cell. Non-limiting examples of vectors include viral vectors, deproteinized DNA or RNA expression vectors, plasmid, cosmid or phage vectors, cationic condensing agent-associated DNA or RNA expression vectors, liposome-encapsulated DNA or RNA expression vectors, and certain eukaryotic cells such as producer cells.

As used herein, the term "expression control sequence" or "control sequence" refers to a polynucleotide sequence necessary to effect expression of the coding sequence to which it is ligated. The nature of such control sequences differs depending on the host organism. For example, in prokaryotes, such control sequences typically include a promoter, a ribosome binding site, and terminators and, in some cases, enhancers. Thus, the term “control sequence” is intended to include at least all components whose presence is essential for expression, and may also include additional components whose presence is beneficial, such as leader sequences.

"Host cell" includes a single cell or cell culture that can be or is the recipient for vectors for incorporating polynucleotide inserts. Host cells include the progeny of a single host cell, and the progeny need not be completely identical (in morphology or complementary genomic DNA) to the original parent cell due to natural, accidental, or intentional mutation. The host cell includes cells transfected in vivo using the polynucleotides of the present invention.

As used herein, “mammalian cells” includes reference to cells derived from mammals, including humans, rats, mice, guinea pigs, chimpanzees or macaques. Cells can be cultured in vivo or in vitro .

As used herein, the term “purified product” refers to a preparation of a product isolated from the cellular constituents with which the product is normally associated and/or from other cell types that may be present in the sample of interest.

As used herein, the term "substantially pure" refers to a material that is at least 50% pure (i.e. free from contaminants), more preferably at least 90% pure, more preferably at least 95% pure % pure, even more preferably at least 98% pure, and most preferably at least 99% pure.

As used herein, the term “malignancy” or “malignant” refers to a physiological condition in mammals typically characterized by unregulated cell growth, neoplasia, or tumor resulting from abnormal, uncontrolled cell growth. In some aspects, the term “malignancy” refers to a malignant primary tumor that has not metastasized and remains localized. In other aspects, the term “malignant neoplasm” refers to a malignant tumor that invades and destroys adjacent body structures and spreads to distant sites. In some aspects, the cancer is associated with a specific cancer antigen. Non-limiting examples of malignancies include malignancies of the oral cavity and pharynx, the digestive system (eg, esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile ducts and pancreas) or endocrine system. In some embodiments, the digestive system malignancy is cancer of the esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile duct, and pancreas.

As used herein, the term “esophageal cancer” is intended to include a generally accepted medical definition defining esophageal cancer as a medical condition characterized by a malignant growth of cells of the esophagus. Examples of esophageal malignancies include adenocarcinoma, squamous cell carcinoma, choriocarcinoma, lymphoma, sarcoma, and small cell carcinoma.

Gastrointestinal (GI) malignancy is a term for a group of malignancies affecting the digestive system. As used herein, the term "stomach cancer" is intended to include a generally accepted medical definition defining gastric cancer as a medical condition characterized by a malignant growth of cells of the stomach. Specifically, stomach cancer is a disease in which malignant cells form in the lining of the stomach. Stomach cancer can develop in any part of the stomach and can spread throughout the stomach and to other organs, particularly the esophagus, lungs and liver.

As used herein, the term “colorectal cancer” is intended to include a generally accepted medical definition defining colorectal cancer as a medical condition characterized by a malignant growth of cells of the gastrointestinal tract below the small intestine (i.e., the large intestine, including the cecum, ascending colon, transverse colon, descending colon, sigmoid colon and rectum). Moreover, for purposes of the invention, the term “colorectal cancer” is intended to include medical conditions characterized by malignancy arising from cells of the duodenum and small intestine (jejunum and ileum). The definition of colorectal cancer used herein is broader than the generally accepted medical definition, but is presented as such because cells of the duodenum and small intestine also contain GUCY2c.

For the purposes of the invention, the term “malignant cell” or “malignant neoplasm” refers to tumors or tumor cells that are invasive and/or capable of metastasizing, i.e. malignant cells.

As used herein, the term “treat” or “treatment” means an approach to achieve beneficial or desired clinical results. For purposes of the present invention, treatment is defined as administering to an individual, eg a patient, an anti-GUCY2c antibody molecule (eg, a monoclonal anti-GUCY2c antibody or a bispecific or polyspecific anti-GUCY2c antibody). Such administration, for example, can be accomplished by direct administration to the individual or by application to an isolated tissue or cell from the individual, which is then returned to the individual's body. The anti-GUCY2c antibody molecule can be administered alone or in combination with one or more agents. The treatment may be intended to cure, heal, alleviate, relieve, modify, restore, improve, temporarily alleviate, or influence a disorder, symptoms of a disorder, or a predisposition to a disorder, such as a cancer. In some embodiments, the treatment is useful for any one or more of the following: (a) treating, preventing, or improving one or more symptoms of a condition associated with GUCY2c-expressing cancer cells in an individual (for example, a gastrointestinal cancer such such as colorectal cancer (CRC)); (b) inhibiting tumor growth or progression in an individual having a cancer expressing GUCY2c; (c) inhibiting metastasis of cancer cells expressing GUCY2c in an individual having one or more cancer cells expressing GUCY2c; (d) inducing regression (eg, long-term regression) of a tumor expressing GUCY2c; (e) inducing cytotoxic activity in malignant cells expressing GUCY2c; (f) improving progression-free survival in an individual with a GUCY2c-associated disorder; (g) improving overall survival of an individual with a GUCY2c-associated disorder; (h) reducing the use of additional chemotherapeutic or cytotoxic agents in an individual with a GUCY2c-associated disorder; (i) reducing the tumor burden in an individual with a GUCY2c-associated disorder; or (j) blocking the interaction of GUCY2c with other as yet unidentified factors. Without wishing to be bound by any theory, it is believed that the treatment causes inhibition, ablation or destruction of a cell in vitro or in vivo or other reduction in the ability of a cell, including an abnormal cell, to mediate a disorder, for example, a disorder described herein, such as cancer neoplasm.

As used herein, the term "antidiarrheal agent" means a drug that stops or slows down diarrhea. In addition to supportive care, which may include fluid administration along with cessation of lactose, alcohol, and high osmolarity foods, these antidiarrheals may be administered with or in combination with anti-GUCY2c bispecific antibodies. Antidiarrheals include, but are not limited to, bismuth subgallate, Lactobacillus acidophilus , Saccharomyces boulardii , loperamide/simethicone, atropine/diphenoxylate, atropine/difenoxin, Saccharomyces boulardii lyophilisate, Lactobacillus acidophilus , loperamide, bismuth subsalicylate, Lactob acillus acidophilus / Lactobacillus bulgaricus , Lactobacillus rhamnosus , attapulgite, crofelemer, fluoroquinolone, antibiotic or octreotide. See Bensen et al. Recommended Guidelines for the Treatment of Cancer Treatment-Induced Diarrhea. Journal of Clinical Oncology 14: 2918-2926, 2004.

As used herein, the term “subject” is intended to include any animal (eg, mammal), including, but not limited to, humans, non-human primates, rodents, and the like, that will be the recipient of a particular treatment. For example, the individual may be a patient (eg, a human patient or a veterinary patient) having a cancer. Generally, the terms “individual” and “patient” are used interchangeably herein to refer to a person.

For purposes of the invention, the term "non-human animals" includes all non-human vertebrates, for example, non-human mammals and non-mammals, such as non-human primates, sheep, dog, cow, chicken, amphibians, reptiles, mouse, rat, rabbit or goat, etc., unless otherwise stated.

As used herein, the term “pharmaceutically acceptable” refers to a product or compound that has been approved (or approved) by a regulatory agency of the federal or state government or listed in the United States Pharmacopoeia or other generally accepted pharmacopoeias for use in animals, including humans.

As used herein, the terms “pharmaceutically acceptable excipient, carrier or excipient” or “acceptable pharmaceutical carrier” refer to an excipient, carrier or excipient that can be administered to a subject along with at least one antibody of the present invention and that does not interfere with the activity of the antibody. The excipient, carrier or excipient must be non-toxic when administered with the antibody in doses sufficient to achieve a therapeutic effect.

As used herein, the term “improvement” means a reduction or improvement of one or more symptoms compared to the absence of administration of the antibody molecule of the invention. The term "improvement" also includes a decrease in the duration of a symptom.

As used herein, the term “prevention” refers to preventing the recurrence or onset of one or more symptoms of a disorder in an individual as a result of the administration of a prophylactic or therapeutic agent.

As used herein, the terms "effective amount", "therapeutically effective amount", "therapeutically sufficient amount" or "effective dose" refer to any amount of a therapeutic agent that is effective or sufficient after administration of a single or multiple doses to an individual for the prevention, healing, improvement or treating a disease, disorder or side effect, or reducing the rate of progression of the disease or disorder, or prolonging the cure, relief or improvement of the condition of an individual with a disorder presented herein beyond what would be expected in the absence of such treatment. The term also includes amounts effective to enhance normal physiological function. An effective amount may be considered in relation to the administration of one or more therapeutic agents, and a single agent may be considered to be administered in an effective amount if the desired result can be or is achieved concurrently with one or more other agents.

As used herein, the term “growth inhibition” of a tumor or malignancy refers to slowing, preventing or stopping growth and/or metastasis and does not necessarily indicate overall elimination of tumor growth.

Potency is a measure of the activity of a therapeutic agent, expressed in terms of the amount required to achieve an effect at a specified intensity. A highly active agent causes a greater response at low concentrations compared to a lower activity agent causing a lesser response at low concentrations. Activity is a function of affinity and potency. The term "efficacy" refers to the ability of a therapeutic agent to induce a biological response upon binding to a target ligand and the quantitative magnitude of this response. As used herein, the term "half-maximal effective concentration (EC ₅₀ )" refers to the concentration of a therapeutic agent that produces a response that is half the value between baseline and the maximum after a specified exposure time. The therapeutic agent may cause inhibition or stimulation. The EC ₅₀ value is a commonly used value and is used herein as a measure of activity.

Within the scope of the invention, the term "combination treatment" or administration "in combination with" refers to the use of more than one prophylactic and/or therapeutic agent. The use of the term “combination treatment” or “in combination” does not limit the order in which the prophylactic and/or therapeutic agents are administered to the individual with the disorder. In other words, combination treatment can be carried out through separate, sequential or simultaneous treatment with therapeutic agents. In the case of "sequential administration", the first agent administered may cause some physiological effect in the individual when the second agent is administered or becomes active in the individual.

As used herein, the term "simultaneous administration" in relation to the administration of a prophylactic and/or therapeutic agent refers to the administration of the agents in such a way that the individual agents are present in the body of an individual at the same time. Coadministration may be affected by molecules formulated in the same composition or separate compositions administered simultaneously or closely in time. If necessary, sequential administration can be carried out in any order.

As used herein, the term "GUCY2c" refers to mammalian guanylate cyclase C (GUCY2c), preferably human GUCY2c protein. The term "GUCY2c" can be used interchangeably with the term "GUCY2C". The nucleotide sequence of human GUCY2c is described as GenBank accession number: NM.sub.-004963, incorporated herein by reference. The amino acid sequence of human GUCY2c is described as GenBank accession number NP.sub.-004954, incorporated herein by reference.

Typically, the naturally occurring allelic variant has an amino acid sequence that is at least 95%, 97%, or 99% identical to the protein represented by GenBank accession number NP.sub.-004954. The GUCY2c protein has been characterized as a cell surface transmembrane receptor protein and is thought to play a critical role in intestinal fluid maintenance, electrolyte homeostasis, and cell proliferation.

As used herein, the terms “GUCY2c-binding antibody,” “GUCY2c-recognizing antibody,” “anti-GUCY2c antibody,” “anti-GUCY2c antibody molecule” include a molecule that combines at least one binding domain of an antibody (as defined herein) ) with at least one anti-GUCY2c antibody binding domain (as defined herein). The term "anti-GUCY2c antibody molecule of the present invention" refers to antibodies and antigen-binding fragments thereof that interact with or recognize, e.g., bind (eg, bind specifically) GUCY2c, e.g., human GUCY2c, mouse GUCY2c, rat GUCY2c, cynomolgus GUCY2c.

The term "cluster of differentiation 3" or "CD3" refers to a multimeric protein complex historically known as the T3 complex and consisting of four distinct polypeptide chains: epsilon (ε), gamma (γ), delta (δ), zeta (ζ), assembled and functioning as three pairs of dimers (εγ, εδ, ζζ). The CD3 complex serves as a T cell co-receptor bound non-covalently to the T cell receptor (TCR) (Smith-Garvin et al. 2009) and generates an activation signal in T lymphocytes. The CD3 protein complex is a defining feature of T cell lineage, thus anti-CD3 antibodies can be effectively used as T cell markers (Chetty and Gatter, Journal of Pathology, Vol 172, 4, 303-301 (1994)). Anti-CD3 antibodies are known to induce the generation of cytotoxic T cells through activation of endogenous lymphokine production and are capable of selectively killing tumor targets (Yun et al., Cancer Research, 49:4770-4774, 1989).

More specifically, T cells express TCR complexes capable of inducing antigen-specific immune responses (Smith-Garvin et al., Annula Review of Immunology, 27: 1, 591-619 (2009)). Antigens are peptides expressed by tumor cells and virus-infected cells that can stimulate immune responses. Intracellularly expressed antigens are bound to major histocompatibility complex class I (MHC class I) molecules and transported to the surface where they are presented to T cells. If the binding affinity of the TCR to MHC class I complexed with antigen is sufficient, the formation of an immune synapse will be initiated. Signal transduction across the immune synapse is mediated by CD3 co-receptors, which form εδ, εγ and ζζ dimers. These dimers bind to the TCR and generate an activation signal in T lymphocytes. This signaling cascade directs T cell-mediated killing of the cell expressing the antigen. Cytotoxicity is mediated by the release and transfer of granzyme B and perforin from the T cell to the target cell.

As used herein, the terms “CD3-binding antibody,” “CD3-recognizing antibody,” “anti-CD3 antibody,” “anti-CD3 antibody molecule” include antibodies and antigen-binding fragments thereof that specifically recognize one subunit of CD3 (e.g., epsilon, delta, gamma or zeta), as well as antibodies and their antigen-binding fragments that specifically recognize the dimeric complex of two CD3 subunits (for example, gamma/epsilon, delta/epsilon and zeta/zeta CD3 dimers). Human CD3 epsilon is listed under GenBank accession number NM_000733. Antibodies and antigen binding fragments of the present invention can bind to soluble CD3 and/or CD3 expressed on the surface of a cell. Soluble CD3 includes natural CD3 proteins as well as recombinant variants of the CD3 protein, such as, for example, monomeric and dimeric CD3 constructs that lack the transmembrane domain or are not otherwise associated with the cell membrane.

Antibodies against CD3 can generate an activation signal in T lymphocytes. Other T cell activating ligands may also be used, including, but not limited to, CD28, CD134, CD137, and CD27.

Anti-CD3 bispecific antibodies bypass the need for MHC peptide/TCR engagement and instead recruit T cells to target cells expressing cell surface antigen. One arm of the bispecific antibody binds to a tumor-associated antigen on the cell surface, and the other arm binds to the CD3 protein, which is part of the TCR complex on T cells. Co-engagement of CD3 on T cells and target antigen on tumor cells via a bispecific antibody results in a cytotoxic response. Thus, without wishing to be bound by any theory, it is believed that the bispecific antibodies of the present invention may allow a T cell to bypass the need for TCR and MHC class I interaction in complex with an antigen and instead redirect T cells to target cells through direct co-involvement of CD3 (such as CD3 epsilon) expressed on the T cell and GUCY2c expressed on the tumor.

As used herein, the term “anti-CD3-GUCY2c bispecific antibody” refers to a molecule designed to direct an individual's T cells to kill cancer cells by targeting tumor cells expressing the desired molecule. In some embodiments, the desired molecule is human GUCY2c. In some embodiments, the anti-CD3-GUCY2c bispecific antibody contains two FV domains. In some embodiments, the anti-CD3-GUCY2c bispecific antibody comprises a first anti-GUCY2c FV domain and a second anti-CD3 FV domain. FV domains may be scFV domains.

For purposes of the invention, a “first polypeptide” is any polypeptide that can be linked to a second polypeptide. The first polypeptide and the second polypeptide are connected in a contact region. In addition to the contact region, the first polypeptide may contain one or more additional domains, such as "binding domains" (eg, an antibody variable domain, receptor binding domain, ligand binding domain, or enzymatic domain) or antibody constant domains (or portions thereof) ), including the CH2, CH1 and CL domains. Typically, the first polypeptide will contain at least one antibody-derived domain. This domain is typically a constant domain, such as the CH3 domain of an antibody, and may form a contact region of the first polypeptide. Examples of the first polypeptides include antibody heavy chain polypeptides, chimeras that combine an antibody constant domain with a heterologous polypeptide binding domain, receptor polypeptides, ligand polypeptides, and antibody variable domain polypeptides (eg, bispecific antibodies).

In addition to the contact region, the second polypeptide may contain additional domains, such as a “binding domain” (e.g., an antibody variable domain, a receptor binding domain, a ligand binding domain, or an enzymatic domain), or antibody constant domains (or portions thereof), including CH2, CH1 and CL domains. Typically, the second polypeptide will contain at least one antibody-derived domain. This domain is typically a constant region, such as the CH3 domain of an antibody, and may form a contact region for a second polypeptide. Examples of second polypeptides include antibody heavy chain polypeptides, chimeras that combine an antibody constant domain with the binding domain of a heterologous polypeptide, and antibody variable domain polypeptides (eg, bispecific antibodies).

As used herein, the terms "complex" or "complexed" refer to the binding of two or more molecules interacting with each other through bonds and/or forces (eg, van der Waals, hydrophobic, hydrophilic forces) other than peptide bonds. In one embodiment, the complex is heteromultimeric. It should be understood that, as used herein, the term “protein complex” or “polypeptide complex” includes complexes comprising a non-proteinaceous substance conjugated to a protein in a protein complex (for example, including, but not limited to, chemical molecules such as a toxin or detection agent) .

Although any materials and methods similar or equivalent to those presented herein may be used in the practice or testing of the present invention, preferred materials and methods will be described below.

Materials and methods

Various methods for producing antibodies have been described, including the conventional hybridoma method for producing monoclonal antibodies, recombinant methods for producing antibodies (including chimeric antibodies, eg, humanized antibodies), producing antibodies in transgenic animals, and the recently described phage display technology for producing “fully human” antibodies. These methods are briefly described below.

Polyclonal antibodies against an antigen of interest can generally be induced in animals through repeated subcutaneous (sc) or intraperitoneal (ip) injections of antigen and adjuvant. The antigen (or fragment containing the target amino acid sequence) can be conjugated to a protein that is immunogenic for the species being immunized, such as serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor, using a bifunctional or derivatizing agent, such as maleimidobenzoylsulfosuccinimide ester (conjugation via cysteine residues) or N-hydroxysuccinimide (via lysine residues). Animals are immunized with immunogenic conjugates or derivatives and after several weeks the animals are boosted by subcutaneous injection at multiple sites. After 7-14 days, blood is taken from the animals and the serum is analyzed for antibody titer. Animals are subjected to booster immunization until the titer plateau is reached. Preferably, the animal is boosted using a conjugate of the same antigen but conjugated to a different protein and/or via a different cross-linking reagent. Conjugates can also be produced in recombinant cell culture as fusion proteins. In addition, aggregating agents such as alum are used to enhance the immune response.

Monoclonal antibodies can be produced from a population of substantially homogeneous antibodies using the hybridoma method first described in Kohler & Milstein, Nature 256:495, 1975, or can be produced by recombinant DNA methods (Cabilly et al., US Pat. No. 4,816,567). In the hybridoma method, a mouse or other suitable animal host is immunized as described above to induce lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes can be immunized in vitro . The lymphocytes are then fused with myeloma cells using a suitable fusion agent such as polyethylene glycol to produce a hybridoma cell (Goding, Monoclonal antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)). The hybridoma cells thus obtained are seeded and grown in a suitable culture medium, preferably containing one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. In addition, hybridoma cells can be grown in vivo as ascites tumors in an animal. The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid or serum by conventional immunoglobulin purification methods, such as, for example, using protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis or affinity chromatography.

Expression of the antibody, antigen-binding antibody fragments, or any antibody construct can be accomplished in suitable prokaryotic or eukaryotic host cells such as CHO cells, NSO cells, SP2/0 cells, HEK293 cells, COS cells, PER.C6 cells, yeast, or E. coli cells and the secreted antibody is isolated and collected from the cells (cell culture supernatant, conditioned cell culture supernatant, cell lysate, or purified total). General methods for producing antibodies are well known in the art and are described, for example, in review articles by Makrides, SC, Protein Expr. Purif. 17:183-202, 1999; Geisse, S., et al., Protein Expr. Purif. 8:271-282, 1986; Kaufman, R. J., Mo. I. Biotechnol. 16:151-160, 2000; Werner, R. G., Drug Res. 48:870-880, 1998. In a specific embodiment, the cell culture is a mammalian cell culture, such as a Chinese hamster cell culture (CHO).

In various embodiments, the isolated antibodies can be subjected to additional purification steps using conventional chromatographic techniques known in the art. Specifically, purification methods include, but are not limited to, affinity chromatography (e.g., protein A affinity chromatography), ion exchange chromatography (e.g., anion exchange chromatography or cation exchange chromatography), hydrophobic interaction chromatography, hydroxyapatite chromatography, gel filtration, and/or dialysis. Among these, the preferred purification method is the use of protein A chromatography. The matrix to which the affinity ligand is attached is most often agarose, but other matrices are available.

Other methods for purifying antibodies are also known in the art, such as ion exchange column fractionation, ethanol precipitation, reverse phase high performance chromatography, ethanol precipitation, reverse phase HPLC, silica chromatography, heparin-Sepharose chromatography, anion exchange or cation exchange chromatography. resin (for example, on a column with polyaspartic acid), chromatofocusing, SDS-PAGE and precipitation with ammonium sulfate. The above list of cleaning methods are examples only and are not intended to be limiting.

Alternatively, it is possible to produce transgenic animals (eg mice) capable of producing the full repertoire of human antibodies after immunization in the absence of endogenous immunoglobulin production. For example, homozygous deletion of the antibody heavy chain junction region gene (J.sub.H) in chimeric and germline mutant mice has been reported to result in complete inhibition of endogenous antibody production. Transfer of the human germline immunoglobulin gene repertoire to such germline mutant mice will result in the production of human antibodies upon antigen stimulation. See, for example, Jakobovits et al., Proc. Natl. Acad. Sci. USA 90:2551-255;1993 and Jakobovits et al., Nature 362:255-258, 1993.

In some embodiments, the antibodies of the present invention can be humanized while maintaining high affinity for the antigen and other favorable biological properties. Methods for humanizing non-human antibodies are well known in the art. Humanization, in essence, can be carried out in the manner of Winter et al. (Jones et al., Nature 321:522-525, 1986; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988) by replacing CDRs or rodent CDR sequences corresponding human antibody sequences. Thus, such humanized antibodies are chimeric antibodies (Cabilly, supra), where a substantially less than intact human variable domain is replaced with the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are replaced with residues from analogous regions in rodent antibodies. It is important to humanize antibodies while maintaining high affinity for the antigen and other favorable biological properties. To achieve this goal, in a preferred method, humanized antibodies are obtained by analyzing the parent sequences and various conceptual humanized products using three-dimensional models of the parent and humanized sequences. Three-dimensional models of immunoglobulins are well known to those skilled in the art. Computer programs are available that illustrate and display possible three-dimensional conformational structures of selected candidate immunoglobulin sequences. The study of such mappings allows one to analyze the possible role of residues in the functioning of the candidate immunoglobulin sequence, i.e. analyze residues that affect the ability of a candidate immunoglobulin to bind an antigen. Thus, FR residues can be selected and combined from the consensus and import sequence in a manner that achieves the desired antibody characteristic, such as increased affinity for target antigens. See WO92/22653 published December 23, 1992 for more details.

Antibodies of the present invention can also be produced using various phage display methods known in the art. In phage display methods, the functional domains of an antibody are displayed on the surface of phage particles bearing polynucleotide sequences encoding them. In a particular aspect, such a phage can be used to display antigen binding domains, such as Fab and Fv or disulfide bond stabilized Fv, expressed from a repertoire or combinatorial library of antibodies (eg, human or mouse). A phage expressing an antigen-binding domain that binds an antigen of interest can be selected for or identified using an antigen, for example, using a labeled antigen or an antigen bound or fixed to a solid surface or particle. The phage used in these methods is typically filamentous phage, including fd and M13. The antigen binding domains are expressed as a recombinant fusion protein with the phage gene III or gene VIII protein. Examples of phage display methods that can be used to produce immunoglobulins or fragments thereof of the present invention include those described in Brinkmann et al. "Phage display of Disulfide-Stabilized Fv Fragments," J. Immunol. Methods, 182:41-50, 1995.

The functional characteristics of multiple IgG isotypes and their domains are well known in the art. The amino acid sequences of IgG1, IgG2, IgG3 and IgG4 are known in the art. The selection and/or combination of two or more domains from specific IgG isotypes for use in the methods of the invention may be based on any known parameter of the parent isotypes, including affinity for FcγR. For example, regions or domains of IgG isotypes that exhibit limited or no binding to FcγRIIB, such as IgG2 or IgG4, can be used if the bispecific antibody is desired to be designed to maximize binding to the activating receptor and minimize binding to the inhibitory receptor. Likewise, the use of Fc chains or domains from IgG isotypes known to bind preferentially to C1q or FcγRIIIA, such as IgG3, can be combined with Fc amino acid modifications known in the art to enhance antibody-dependent cellular cytotoxicity (ADCC) ) and/or complement-mediated cytotoxicity (CDC), to design a bispecific antibody in a manner that maximizes the activity of an effector function, such as complement activation or ADCC. Likewise, mutations can be generated in the Fc chains or domains of IgG isotypes that minimize or eliminate the effector function of the Fc chain.

During the discovery process, information about the desired epitopes can be obtained through the production and characterization of antibodies. This information can then be used to competitively screen antibodies for binding to the same epitope. An approach to achieve this is to perform competition and cross-competition studies to detect antibodies that compete or cross-compete with each other, for example, antibodies that compete for binding to an antigen. One method is to identify the epitope to which antibodies bind, or "epitope mapping." There are many methods known in the art for mapping and characterizing the localization of epitopes on proteins, including crystal structure resolution of the antibody-antigen complex, competition assays, gene fragment expression assays, and synthetic peptide-based assays, as described, for example, in Chapter 11 of Harlow and Lane. Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1999. In an additional example, epitope mapping can be used to determine the sequence to which an antibody binds. Epitope mapping is commercially available from various sources, such as Pepscan Systems (Edelhertweg 15, 8219 PH Lelystad, The Netherlands). In a further example, antigen binding domain mutagenesis, domain swapping experiments, and alanine scan mutagenesis can be performed to identify residues required, sufficient, and/or essential for epitope binding. The binding affinity and reverse reaction rate of the antigen binding domain can be determined using competitive binding assays. One example of a competitive binding assay is a radioimmunoassay, which involves incubating a labeled antigen and detecting a molecule bound to the labeled antigen. The affinity of the molecule of the present invention for the antigen and the rate of reverse binding reaction can be determined from saturation data in the Scatchard assay.

The affinities and binding properties of the antibodies of the present invention for an antigen can initially be determined using in vitro assays (biochemical or immunoassays) known in the art for the antigen binding domain, including, but not limited to, enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), biolayer interferometry or immunoprecipitation assays. The molecules of the invention may have binding properties in in vivo models (such as those described and presented herein) similar to those in in vitro assays. However, the present invention does not exclude molecules of the invention that do not exhibit the desired phenotype in in vitro assays but exhibit the desired phenotype in vivo .

An optional bispecific antibody format is the Fv strategy based on a covalently linked, bispecific heterodimeric diabody structure, also known as dual affinity retargeting antibody ( ^DART® ), described, for example, in US Patent Publications Nos. 2007/0004909, 2009/0060910 and 2010 /0174053).

Once the nucleic acid sequence encoding the molecules of the invention (ie, the binding domains) has been obtained, a vector for producing the molecules can be prepared by recombinant DNA technology using methods well known in the art.

Polynucleotides encoding the binding domains of an antibody (e.g., anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) of the present invention may include an expression control polynucleotide sequence operably linked to antibody coding sequences, including naturally occurring or heterologous promoter regions known in the art. areas. Expression control sequences can be eukaryotic promoter systems in vectors that can transform or transfect eukaryotic host cells, but control sequences for prokaryotic hosts can also be used. After insertion of the vector into a suitable host cell line, the host cell is grown under conditions suitable for expression of nucleotide sequences and, if desired, collection and purification of antibodies. Eukaryotic cell lines include CHO cell lines, various COS cell lines, HeLa cells, myeloma cell lines, transformed B cells or human embryonic kidney cell lines.

In one embodiment, the DNA encoding the antibodies of the invention is isolated and sequenced by conventional methods (eg, using oligonucleotide probes capable of specifically binding to the genes encoding the heavy and light chains of mouse antibodies). The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA can be inserted into expression vectors, which are then transfected into host cells, such as monkey COS cells, CHO cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to synthesize monoclonal antibodies in recombinant host cells. The DNA can also be modified, for example, by replacing the coding sequence of the human heavy and light chain constant domains with homologous sequences in mice, Morrison et al., Proc. Nat. Acad. Sci. 81:6851, 1984. Thus, chimeric antibodies are produced having the binding specificity of a monoclonal antibody against the antigen of the present invention.

As a cell surface protein, GUCY2c can serve as a therapeutic target for receptor-binding proteins such as antibodies or ligands. In normal intestinal tissue, GUCY2c is expressed on the apical side of the tight junction of epithelial cells that form the barrier between the luminal environment and the vascular compartment (Almenoff et al., Mol Microbiol 8: 865-873, 1993; and Guarino et al., Dig Dis Sci 32: 1017 -1026, 1987). In this regard, systemic intravenous administration of a GUCY2c-binding protein therapeutic would have minimal effect on intestinal GUCY2c receptors present on normal cells while allowing access to neoplastic cells of the gastrointestinal tract, including malignant or metastatic colon cancer cells. , extraintestinal or metastatic colon tumors, esophageal tumors, gastric tumors or adenocarcinoma at the gastroesophageal junction. In addition, GUCY2c is internalized through receptor-mediated endocytosis after ligand binding (Buc et al. Eur J Cancer 41: 1618-1627, 2005; Urbanski et al., Biochem Biophys Acta 1245: 29-36, 1995).

Tissue-specific expression and association of GUCY2c with malignancy, such as those of gastrointestinal origin (eg, colon cancer, gastric cancer, or esophageal cancer), can be used to use GUCY2c as a diagnostic marker for this disease (Carrithers et al., Dis Colon Rectum 39 :171-181, 1996; Buc et al. Eur J Cancer 41: 1618-1627, 2005).

The present invention provides an antibody that binds to GUCY2c (e.g., human GUCY2c (e.g., SEQ ID NO: 224, derived from the protein accession number: NP_004954.2)), and which can be used in any one or more of the following: (a ) treating, preventing, or improving one or more symptoms of a condition associated with GUCY2c-expressing cancer cells in an individual (eg, gastrointestinal cancer, such as colorectal cancer (CRC)); (b) inhibiting tumor growth or progression in an individual having a cancer expressing GUCY2c; (c) inhibiting metastasis of cancer cells expressing GUCY2c in an individual having one or more cancer cells expressing GUCY2c; (d) inducing regression (eg, long-term regression) of a tumor expressing GUCY2c; (e) causing cytotoxic activity against malignant cells expressing GUCY2c; (f) improving progression-free survival in an individual with a GUCY2c-associated disorder; (g) improving overall survival of an individual with a GUCY2c-associated disorder; (h) reducing the use of additional chemotherapeutic or cytotoxic agents in an individual with a GUCY2c-associated disorder; (i) reducing the tumor burden in an individual with a GUCY2c-associated disorder; or (j) blocking the interaction of GUCY2c with other as yet unidentified factors.

In one aspect, the present invention provides an antibody comprising (a) a heavy chain variable region (VH) comprising a VH complementarity determining region one (VH CDR1), a VH complementarity determining region two (VH CDR2) and a VH complementarity determining region three (VH CDR3). VH) VH sequence given in SEQ ID NO: 11, 19, 26, 33, 41, 48, 52, 57, 60, 62, 64, 65, 67, 69, 71 or 73; and/or (b) a light chain variable region (VL) comprising a VL complementarity determining region one (VL CDR1), a VL complementarity determining region two (VL CDR2) and a VL complementarity determining region three (VL CDR3) of the VL sequence set forth in SEQ ID NO: 92, 100, 104, 106, 112, 119, 125, 129, 134, 136, 137, 138, 140, 143, 145, 147, 150, 152, 156, 158, 160, 162, 166, 170 , 171, 172, 173, 174 or 175.

In another aspect, the present invention provides an antibody having any of the partial heavy chain sequences and/or any of the partial light chain sequences set forth in Table 1. In one embodiment, the present invention provides an antibody comprising a VH region and/or a VL region where: (a) the VH region contains SEQ ID NO: 11, 19, 26, 33, 41, 48, 52, 57, 60, 62, 64, 65, 67, 69, 71 or 73; and/or (b) the VL region contains SEQ ID NO: 92, 100, 104, 106, 112, 119, 125, 129, 134, 136, 137, 138, 140, 143, 145, 147, 150, 152, 156 , 158, 160, 162, 166, 170, 171, 172, 173, 174 or 175.

Table 1

Description Subsequence GUCY2C-0074_VH EVQLQQSGAELARPGASVNLSCKAS GYTFT TYWMQ WVKQRPGQGLEWIG AI YPGDGM TTYTQKFKD KATLTADKSSSTAYMQLSSLASEDSAVYYCVR KGMDY WGQGTSVTVSS (SEQ ID NO: 11) GUCY2C-0077_VH EVQLQQSGAELARPGASVKLSCKAS GYTFT KYWMQ WIKQRPGQGLEWIG AI YPGDGF TTYTQKFKG KATLTADKSSNTAYMQLSSLASEDSAVYYCAR RNYGRTYGGDY WGQGTSVTVSS (SEQ ID NO: 19) GUCY2C-0098_VH QVQLQQPGAELVKPGASVKLSCKAS GYTFT SYWMH WVKQRPGQGLEWIG EI KPSNGL TNYIEKFKN KATLTVDKSATTAYMQLSSLTAEDSAVYYCTR TITTTEGYWFFDV WGAGTTVTVSS (SEQ ID NO: 26) GUCY2C-0104_VH EVQLQQSGAELVKPGASVKLSCTAS GFNIK DTYIH WVKQRPEQGLEWIG RI DPANGN ANYDPKFQG KATITADTSSNTAYLQLSSLTSEDTAVFYCSS LGTGTY WGQGTTLTVSS (SEQ ID NO: 33) GUCY2C-0105_VH EVQLQQSGPELVKPGASVKISCKAS GYSFT DYIML WVKQSHGKSLEWIG NS NPYYGS TSYNLKFKG KATLTVDKSSSTAYMHLNSLTSEDSAVYYCAR SGYYGSSPYWYFDV WGAGTTVTVSS (SEQ ID NO: 41) GUCY2C-0240_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT TYWMQ WVRQAPGKGLEWIG AI YPGDGM TTYTQKFKD RFTISADKAKNSAYLQMNSLRAEDTAVYYCVR KGMDY WGQGTLVTVSS
(SEQ ID NO: 48) GUCY2C-0315_VH QVQLVQSGAEVKKPGASVKVSCKAS GYTFT TYWMQ WVRQAPGQGLEWIG AI YPGDGM TTYTQKFQG RVTMTRDTSTSTVYMELSSLRSEDTAVYYCVR KGMDY WGQGTLVTVSS (SEQ ID NO: 52) GUCY2C-0179_VH EVQLVESGGGLVQPGGSLRLSCAAS GYSFT DYIML WVRQAPGKGLEWIG NS NPYYGS TSYNLKFKG RFTISVDKAKNSAYLQMNSLRAEDTAVYYCAR SGYYGSSPYWYFDV WGQGTMVTVSS (SEQ ID NO: 57) GUCY2C-0193_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIG EI KPSNGL TNYIEKFKN RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 60) GUCY2C-0210_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIA EI KPSNGL TNYIEKFKN RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 62) GUCY2C-0212_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIG EI KPSNGL TNYIEKFKN RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 64) GUCY2C-1186_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIG EI KPSNGL TNIHPKFKN RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 65) GUCY2C-1476_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIG EI KPSNGL TNVHEKFKN RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 67) GUCY2C-1478_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIG EI KPSNGL TNVHEKFKN RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 69) GUCY2C-1512_VH EVQLVESGGGLVQPGGSLRLSCAAS GYTFT SYWMH WVRQAPGKGLEWIG EI KPSNGL TNYAEQFKN RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 71) GUCY2C-1554_VH; GUCY2C-1608_VH EVQLVESGGGLVQPGGSLRLSCAAS GFTFS SYWMH WVRQAPGKGLEWIG EI KPSNEL TNVHEKFKD RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR TITTTEGYWFFDV WGQGTLVTVSS (SEQ ID NO: 73) GUCY2C-0074_VL DIVLTQSPASLAVSLGQRATISC RASESVDNYGISFMN WFQQKPGQPPKLLIY AASNPGS GVPARFSGSGSGTDFSLNIHPLEEDDTAMFFC QQSKEVPYT FGGGTKLEIK (SEQ ID NO: 92) GUCY2C-0077_VL DIVLTQSPASLAVSLGQRATISC RASESVDNFDISFMN WFQQKPGQPPKLLIY AASNQGS GVPARFSGSGSGTDFSLNIHPMEEDDTAMYFC QQSKEVPYT FGGGTKLEIK (SEQ ID NO: 100) GUCY2C-0078_VL DIVLTQSPASLAVSLGQRATISCRAGESVDNFDISFMNWFQQKPGQPPKLLIY AASNQGS GVPARFSGSGSGTDFSLNIHPMEEDDTAMYFC QQSKEVPYT FGGGTKLEIK (SEQ ID NO: 104) GUCY2C-0098_VL DIVLTQSPASLAVSLGQRATISC RASESVDYYGTSLMQ WYQQKPGQPPKLLIY AASNVES GVPARFSGSGSGTDFSLNIHPVEEDDIAMYFC QQTRKVYT FGGGTKLEIK (SEQ ID NO: 106) GUCY2C-0104_VL DIVMTQSPASLAVSLGQRATISC RASKGVTTSGYSYMH WYQQKPGQPPKLLIY LASNLES GVPARFSGSGSGTDFTLNIHPVEEEDAATYYC QHSREFPLT FGAGTKLELK (SEQ ID NO: 112) GUCY2C-0105_VL DIVMTQSPSSLAVSVGEKVTVSC KSSQSLLYSSNQKNYLA WYQQRPGQSPKLLIY WASTRES GVPDRFTGSGSGTDFTLSISSVKAEDLAVYYC QQYYSYPT FGGGTKLEIK (SEQ ID NO: 119) GUCY2C-0240_VL DIQLTQSPSSLSSASVGDRVTITC RASESVDNYGISFMN WFQQKPGKAPKLLIY AASNPGS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQSKEVPYT FGQGTKLEIK (SEQ ID NO: 125) GUCY2C-0315_VL EIVLTQSPATLSLSPGERATLSC RASESVDNYGISFMN WYQQKPGQAPRLLIY AASNPGS GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQSKEVPYT FGQGTKVEIK (SEQ ID NO: 129) GUCY2C-0179_VL DIQMTQSPSSLSSASVGDRVTITC KSSQSLLYSSNQKNYLA WYQQKPGKSPKLLIY WASTRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYSYPT FGGGTKVEIK (SEQ ID NO: 134) GUCY2C-0193_VL DIQMTQSPSSLSSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASNVES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKVYT FGQGTKLEIK (SEQ ID NO: 136) GUCY2C-0210_VL DIQLTQSPSSSLSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASNVES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKVYT FGQGTKLEIK (SEQ ID NO: 137) GUCY2C-0247_VL DIQLTQSPSSLSSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKPPKLLIY AASNVES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKVYT FGQGTKLEIK (SEQ ID NO: 138) GUCY2C-1186_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASKRYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 140) GUCY2C-1467_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASKLWS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKVYT FGQGTKLEIK (SEQ ID NO: 143) GUCY2C-1476_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASKVAP GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 145) GUCY2C-1478_VL; GUCY2C-1608_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGSSLLQ WYQQKPGKAPKLLIY AASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 147) GUCY2C-1481_VL DIQLTQSPSSLSSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASNIAP GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 150) GUCY2C-1512_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGSSLMQ WYQQKPGKAPKLLIY AASKRYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKEYT FGQGTKLKIK (SEQ ID NO: 152) GUCY2C-1518_VL DIQLTQSPSSLSSASVGDRVTITC RASESVDYYGSSLMQ WYQQKPGKAPKLLIY AASNVAP GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 156) GUCY2C-1526_VL DIQLTQSPSSLSSASVGDRVTITC RASESVDYYGSSLMQ WYQQKPGKAPKLLIY AASHRAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 158) GUCY2C-1527_VL DIQLTQSPSSLSSASVGDRVTITC RASESVDYYGTSLMQ WYQQKPGKAPKLLIY AASNVAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKEYT FGQGTKLEIK (SEQ ID NO: 160) GUCY2C-1538_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGHSLMQ WYQQKPGKAPKLLIY AASNRYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYS FGQGTKLEIK (SEQ ID NO: 162) GUCY2C-1554_VL DIQMTQSPSSLSASVGDRVTITC RASQSVDYYGSSLLQ WYQQKPGKAPKLLIY DASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQERKAYT FGQGTKLEIK (SEQ ID NO: 166) GUCY2C-1555_VL DIQMTQSPSSLSASVGDRVTITC RASQSVDYYGSSLLQ WYQQKPGKAPKLLIY AASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQERKAYT FGQGTKLEIK (SEQ ID NO: 170) GUCY2C-1556_VL DIQMTQSPSSLSASVGDRVTITC RASQSVDYYGSSLLQ WYQQKPGKAPKLLIY DASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 171) GUCY2C-1557_VL DIQMTQSPSSLSASVGDRVTITC RASQSVDYYGSSLLQ WYQQKPGKAPKLLIY AASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 172) GUCY2C-1590_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGSSLLQ WYQQKPGKAPKLLIY DASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQERKAYT FGQGTKLEIK (SEQ ID NO: 173) GUCY2C-1591_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGSSLLQ WYQQKPGKAPKLLIY AASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQERKAYT FGQGTKLEIK (SEQ ID NO: 174) GUCY2C-1592_VL DIQLTQSPSSLSASVGDRVTITC RASESVDYYGSSLLQ WYQQKPGKAPKLLIY DASKLAS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQTRKAYT FGQGTKLEIK (SEQ ID NO: 175)

In Table 1, the underlined sequences are Kabat CDR sequences and the bold ones are Chothia sequences. AbM determination was used for CDR1 VH.

The invention also relates to CDR portions of anti-GUCY2c antibodies. CDR contact regions are regions of an antibody that give the antibody specificity for an antigen. In general, CDR contact regions include positions of residues in the CDR and Vernier regions restricted to maintain the correct loop structure so that the antibody binds to a specific antigen. See, for example, Makabe et al., J. Biol. Chem., 283:1156-1166, 2007. The definition of CDR regions is well known in the field. The algorithm embodied in version 2.3.3 of Abysis (www.abysis.org) was used in the present invention to assign Kabat numbering to the variable regions, CDR1 VL, CDR2 VL, CDR3 VL, CDR2 VH and CDR3 VH, excluding CDR1 VH. AbM determination was used for CDR1 VH.

In one aspect, the present invention provides an antibody having any of the VH CDR sequences and/or any of the VL CDR sequences set forth in Table 2. In one aspect, the present invention provides an antibody that specifically binds guanylate cyclase C (GUCY2c), wherein the antibody contains: (a) a heavy chain variable region (VH) containing (i) a VH complementarity determining region one (VH CDR1) containing SEQ ID NO: 12, 20, 27, 34, 42, 74, 257, 258, 259 , 260 or 261; (ii) a VH complementarity determining region two (VH CDR2) containing SEQ ID NO: 13, 21, 28, 35, 43, 53, 66, 68, 70, 72, 75, 262, 263, 264, 265, 266 or 267; and (iii) VH complementarity determining region three (VH CDR3) containing SEQ ID NO: 14, 22, 29, 36 or 44; and/or (b) a light chain variable region (VL) comprising (i) a VL complementarity determining region one (VL CDR1) comprising SEQ ID NO: 93, 101, 105, 107, 113, 120, 148, 153, 163 or 167; (ii) complementarity determining region VL two (VL CDR2) containing SEQ ID NO: 78, 94, 102, 108, 114, 141, 144, 146, 149, 151, 157, 159, 161, 164 or 168; and (iii) complementarity determining region VH three (CDR3 VL) containing SEQ ID NO: 95, 109, 115, 121, 142, 154, 165 or 169.

In a specific embodiment, the present invention provides an antibody wherein: (a) the VH region comprises (i) a VH CDR1 comprising the sequence SEQ ID NO: 74 or 259; (ii) CDR2 VH containing the sequence SEQ ID NO: 75 or 267; and iii) CDR3 VH containing the sequence SEQ ID NO: 29; and/or (b) the VL region contains (i) a VL CDR1 containing the sequence of SEQ ID NO: 148; (ii) CDR2 VL containing the sequence SEQ ID NO: 149; and (iii) CDR3 VL containing the sequence SEQ ID NO: 142.

table 2

Heavy chain mAb CDR1 VH CDR2 VH CDR3 VH GUCY2C-0074_VH GYTFTTYWMQ (SEQ ID NO: 12) (ABM);
TYWMQ (SEQ ID NO: 257) (Kabat) AIYPGDGMTTYTQKFKD (SEQ ID NO: 13) Kabat);
YPGDGM (SEQ ID NO: 262) (Chothia) KGMDY (SEQ ID NO: 14) GUCY2C-0077_VH GYTFTKYWMQ(SEQ ID NO: 20) (ABM);
KYWMQ (SEQ ID NO: 258) (Kabat) AIYPGDGFTTYTQKFKG (SEQ ID NO: 21) (Kabat);
YPGDGF (SEQ ID NO: 263) (Chothia) RNYGRTYGGDY (SEQ ID NO: 22) GUCY2C-0098_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM); SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNYIEKFKN (SEQ ID NO: 28) (Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-0104_VH GFNIKDTYIH (SEQ ID NO: 34) (ABM);
DTYIH (SEQ ID NO: 260) (Kabat) RIDPANGNANYDPKFQG (SEQ ID NO: 35) (Kabat);
DPANGN (SEQ ID NO: 265) (Chothia) LGTGTY (SEQ ID NO: 36) GUCY2C-0105_VH GYSFTDYIML (SEQ ID NO: 42) (ABM);
DYIML (SEQ ID NO: 261) (Kabat) NSNPYYGSTSYNLKFKG (SEQ ID NO: 43) (Kabat);
NPYYGS (SEQ ID NO: 266) (Chothia) SGYYGSSPYWYFDV (SEQ ID NO: 44) GUCY2C-0240_VH GYTFTTYWMQ (SEQ ID NO: 12) (ABM);
TYWMQ(SEQ ID NO: 257) (Kabat) AIYPGDGMTTYTQKFKD (SEQ ID NO: 13) (Kabat);
YPGDGM (SEQ ID NO: 262) (Chothia) KGMDY (SEQ ID NO: 14) GUCY2C-0315_VH GYTFTTYWMQ (SEQ ID NO: 12) (ABM);
TYWMQ(SEQ ID NO: 257) (Kabat) AIYPGDGMTTYTQKFQG (SEQ ID NO: 53) (Kabat);
YPGDGM(SEQ ID NO: 262) (Chothia) KGMDY (SEQ ID NO: 14) GUCY2C-0179_VH GYSFTDYIML (SEQ ID NO: 42) (ABM);
DYIML (SEQ ID NO: 261) (Kabat) NSNPYYGSTSYNLKFKG(SEQ ID NO: 43) (Kabat);
NPYYGS(SEQ ID NO: 266) (Chothia) SGYYGSSPYWYFDV (SEQ ID NO: 44) GUCY2C-0193_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNYIEKFKN (SEQ ID NO: 28) (Kabat);
KPSNGL(SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-0210_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNYIEKFKN (SEQ ID NO: 28) (Kabat);
KPSNGL(SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-0212_VH GYTFTSYWMH(SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNYIEKFKN(SEQ ID NO: 28) (Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-1186_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNIHPKFKN (SEQ ID NO: 66) (Kabat);
KPSNGL(SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-1476_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNYNEKFKN (SEQ ID NO: 68) (Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-1478_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNVHEKFKN (SEQ ID NO: 70) (Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-1512_VH GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) EIKPSNGLTNYAEQFKN (SEQ ID NO: 72) (Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) GUCY2C-1554_VH; GUCY2C-1608_VH GFTFSSYWMH (SEQ ID NO: 74) (ABM);
SYWMH(SEQ ID NO: 259) (Kabat) EIKPSNELTNVHEKFKD (SEQ ID NO: 75) (Kabat);
KPSNEL (SEQ ID NO: 267) (Chothia) TITTTEGYWFFDV (SEQ ID NO: 29) Light chain mAb CDR1 VL CDR2 VL CDR3 VL GUCY2C-0074_VL RASESVDNYGISFMN (SEQ ID NO: 93) AASNPGS (SEQ ID NO: 94) QQSKEVPYT (SEQ ID NO: 95) GUCY2C-0077_VL RASESVDNFDISFMN (SEQ ID NO: 101) AASNQGS (SEQ ID NO: 102) QQSKEVPYT (SEQ ID NO: 95) GUCY2C-0078_VL RAGESVDNFDISFMN (SEQ ID NO: 105) AASNQGS (SEQ ID NO: 102) QQSKEVPYT (SEQ ID NO: 95) GUCY2C-0098_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASNVES (SEQ ID NO: 108) QQTRKVYT (SEQ ID NO: 109) GUCY2C-0104_VL RASKGVTTSGYSYMH (SEQ ID NO: 113) LASNLES (SEQ ID NO: 114) QHSREFPLT (SEQ ID NO: 115) GUCY2C-0105_VL KSSQSLLYSSNQKNYLA (SEQ ID NO: 120) WASTRES (SEQ ID NO: 78) QQYYSYPT (SEQ ID NO: 121) GUCY2C-0240_VL RASESVDNYGISFMN (SEQ ID NO: 93) AASNPGS (SEQ ID NO: 94) QQSKEVPYT (SEQ ID NO: 95) GUCY2C-0315_VL RASESVDNYGISFMN (SEQ ID NO: 93) AASNPGS (SEQ ID NO: 94) QQSKEVPYT (SEQ ID NO: 95) GUCY2C-0179_VL KSSQSLLYSSNQKNYLA (SEQ ID NO: 120) WASTRES (SEQ ID NO: 78) QQYYSYPT (SEQ ID NO: 121) GUCY2C-0193_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASNVES (SEQ ID NO: 108) QQTRKVYT (SEQ ID NO: 109) GUCY2C-0210_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASNVES (SEQ ID NO: 108) QQTRKVYT (SEQ ID NO: 109) GUCY2C-0247_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASNVES (SEQ ID NO: 108) QQTRKVYT (SEQ ID NO: 109) GUCY2C-1186_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASKRYS (SEQ ID NO: 141) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1467_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASKLWS (SEQ ID NO: 144) QQTRKVYT (SEQ ID NO: 109) GUCY2C-1476_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASKVAP (SEQ ID NO: 146) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1478_VL RASESVDYYGSSLLQ (SEQ ID NO: 148) AASKLAS (SEQ ID NO: 149) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1481_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASNIAP (SEQ ID NO: 151) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1512_VL RASESVDYYGSSLMQ (SEQ ID NO: 153) AASKRYS (SEQ ID NO: 141) QQTRKEYT (SEQ ID NO: 154) GUCY2C-1518_VL RASESVDYYGSSLMQ (SEQ ID NO: 153) AASNVAP (SEQ ID NO: 157) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1526_VL RASESVDYYGSSLMQ (SEQ ID NO: 153) AASHRAS (SEQ ID NO: 159) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1527_VL RASESVDYYGTSLMQ (SEQ ID NO: 107) AASNVAS (SEQ ID NO: 161) QQTRKEYT (SEQ ID NO: 154) GUCY2C-1538_VL RASESVDYYGHSLMQ (SEQ ID NO: 163) AASNRYS (SEQ ID NO: 164) QQTRKAYS (SEQ ID NO: 165) GUCY2C-1554_VL RASQSVDYYGSSLLQ (SEQ ID NO: 167) DASKLAS (SEQ ID NO: 168) QQERKAYT (SEQ ID NO: 169) GUCY2C-1555_VL RASQSVDYYGSSLLQ (SEQ ID NO: 167) AASKLAS (SEQ ID NO: 149) QQERKAYT (SEQ ID NO: 169) GUCY2C-1556_VL RASQSVDYYGSSLLQ (SEQ ID NO: 167) DASKLAS (SEQ ID NO: 168) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1557_VL RASQSVDYYGSSLLQ (SEQ ID NO: 167) AASKLAS (SEQ ID NO: 149) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1590_VL RASESVDYYGSSLLQ (SEQ ID NO: 148) DASKLAS (SEQ ID NO: 168) QQERKAYT (SEQ ID NO: 169) GUCY2C-1591_VL RASESVDYYGSSLLQ (SEQ ID NO: 148) AASKLAS (SEQ ID NO: 149) QQERKAYT (SEQ ID NO: 169) GUCY2C-1592_VL RASESVDYYGSSLLQ (SEQ ID NO: 148) DASKLAS (SEQ ID NO: 168) QQTRKAYT (SEQ ID NO: 142) GUCY2C-1608_VL RASESVDYYGSSLLQ (SEQ ID NO: 148) AASKLAS (SEQ ID NO: 149) QQTRKAYT (SEQ ID NO: 142)

In one aspect, the present invention provides an antibody having any of the partial light chain sequences and/or any of the partial heavy chain sequences set forth in Table 3. In one embodiment, the present invention provides an isolated antibody or antigen binding fragment thereof that specifically binds with CD3, where the antibody contains: a VH region containing CDR1 VH, CDR2 VH and CDR3 VH of the VH sequence shown in SEQ ID NO: 1, 9 or 273; and/or a VL region containing VL CDR1, VL CDR2 and VL CDR3 of the VL sequence set forth in SEQ ID NO: 76, 84, 90, 274, 275 or 276.

Table 3

Description Subsequence VH CD3-0001 EVQLVESGGGLVQPGGSLRLSCAAS GFTFS DYYMT WVRQAPGKGLEWVA FI RNRARGYT SDHNPSVKG RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR DRPSYYVLDY WGQGTTVTVSS (SEQ ID NO: 1) VH CD3-0006 EVQLVESGGGLVQPGGSLRLSCAAS GFTFS DYYMT WVRQAPGKGLEWVA FI RNQARGYT SDHNPSVKG RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR DRPSYYVLDY WGQGTTVTVSS (SEQ ID NO: 9) VH 2B5-1038,
VH 2B5-1039,
VH 2B5-1040 EVQLVESGGGLVQPGGSLRLSCAAS GFTFS DY YMT WVRQAPGKGLEWVA FI RNQARGYT SDHNPSVKG RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR DRPSYYVLDY WGQGTTVTVSS (SEQ ID NO: 273) VL CD3-0001 DIQMTQSPSSLSASVGDRVTITC TSSQSLFNVRSRKNYLA WYQQKPGKAPKLLIY WASTRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK (SEQ ID NO: 76) VL CD3-0004 DIVMTQSPDSLAVSLGERATINC KKSSQSLFNVRSRKNYLA WYQQKPGQPPKLLIS WASTRES GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC KQSYDLFT FGSGTKLEIK (SEQ ID NO: 84) VL CD3-0006 DIQMTQSPSSLSSASVGDRVTITC TSSQSLFNVRSQKNYLA WYQQKPGKAPKLLIY WASTRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK (SEQ ID NO: 90) VL 2B5-1038 DIQMTQSPSSLSASVGDRVTITC TSDQSLFNVRSGKNYLA WYQQKPGKAPKLLIY WASDRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK (SEQ ID NO: 274) VL 2B5-1039 DIQMTQSPSSLSSASVGDRVTITC TSSESLFNVRSGKNYLA WYQQKPGKAPKLLIY WASDRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK (SEQ ID NO: 275) VL 2B5-1040 DIQMTQSPSSLSASVGDRVTITC TSSQSLFNVRSGKNYLA WYQQKPGKAPKLLIY WASDRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK (SEQ ID NO: 276)

In Table 3, the underlined sequences are Kabat CDR sequences and the bold ones are Chothia sequences. AbM determination was used for CDR1 VH.

Table 4 provides examples of CDR sequences for anti-CD3 antibodies presented herein.

Table 4

Heavy chain mAb CDR1 VH CDR2 VH CDR3 VH CD3-0001_VH GFTFSDYYMT (SEQ ID NO: 2) (ABM);
DYYMT(SEQ ID NO: 268) (Kabat) FIRNRARGYTSDHNPSVKG (SEQ ID NO: 3) (Kabat);
RNRARGYT (SEQ ID NO: 269) (Chothia) DRPSYYVLDY (SEQ ID NO: 4) CD3-0006_VH GFTFSDYYMT (SEQ ID NO: 2) (ABM);
DYYMT(SEQ ID NO: 268) (Kabat) FIRNQARGYTSDHNPSVKG(SEQ ID NO: 10) (Kabat);
RNQARGYT (SEQ ID NO: 270) (Chothia) DRPSYYVLDY (SEQ ID NO: 4) 2B5-1038 DYYMT (SEQ ID NO: 268) (Kabat)
GFTFSDY (SEQ ID NO: 277) (Chothia)
GFTFSDYYMT (SEQ ID NO: 2) (extended) FIRNQARGYTSDHNPSVKG (SEQ ID NO: 10) (Kabat)
RNQARGYT (SEQ ID NO: 270) (Chothia) DRPSYYVLDY (SEQ ID NO: 4) 2B5-1039 DYYMT (SEQ ID NO: 268) (Kabat)
GFTFSDY (SEQ ID NO: 277) (Chothia)
GFTFSDYYMT (SEQ ID NO: 2) (extended) FIRNQARGYTSDHNPSVKG (SEQ ID NO: 10) (Kabat)
RNQARGYT (SEQ ID NO: 270) (Chothia) DRPSYYVLDY (SEQ ID NO: 4) 2B5-1040 DYYMT (SEQ ID NO: 268) (Kabat)
GFTFSDY (SEQ ID NO: 277) (Chothia)
GFTFSDYYMT
(SEQ ID NO: 2) (extended) FIRNQARGYTSDHNPSVKG (SEQ ID NO: 10) (Kabat)
RNQARGYT (SEQ ID NO: 270) (Chothia) DRPSYYVLDY (SEQ ID NO: 4) Light chain mAb CDR1 VL CDR2 VL CDR3 VL CD3-0001_VL TSSQSLFNVRSRKNYLA (SEQ ID NO: 77) WASTRES (SEQ ID NO: 78) KQSYDLFT (SEQ ID NO: 79) CD3-0004_VL KSSQSLFNVRSRKNYLA
(SEQ ID NO: 85) WASTRES (SEQ ID NO: 78) KQSYDLFT (SEQ ID NO: 79) CD3-0006_VL TSSQSLFNVRSQKNYLA (SEQ ID NO: 91) WASTRES (SEQ ID NO: 78) KQSYDLFT (SEQ ID NO: 79) 2B5-1038 TSDQSLFNVRSGKNYLA (SEQ ID NO: 278) WASDRES (SEQ ID NO: 281) KQSYDLFT (SEQ ID NO: 79) 2B5-1039 TSSESLFNVRSGKNYLA (SEQ ID NO: 279) WASDRES (SEQ ID NO: 281) KQSYDLFT (SEQ ID NO: 79) 2B5-1040 TSSQSLFNVRSGKNYLA (SEQ ID NO: 280) WASDRES (SEQ ID NO: 281) KQSYDLFT (SEQ ID NO: 79)

In one embodiment, the present invention provides a bispecific antibody wherein: (a) the anti-GUCY2c VL CDR1 comprises the sequence SEQ ID NO: 93, 101, 105, 107, 113, 120, 148, 153, 163, or 167; CDR2 VL against GUCY2c contains the sequence SEQ ID NO: 78, 94, 102, 108, 114, 141, 144, 146, 149, 151, 157, 159, 161, 164 or 168; and CDR3 VL against GUCY2c contains the sequence SEQ ID NO: 95, 109, 115, 121, 142, 154, 165 or 169; (b) the anti-CD3 VH CDR1 contains the sequence SEQ ID NO: 2, 268 or 277; The anti-CD3 VH CDR2 contains the sequence SEQ ID NO: 3, 10, 269 or 270; and the anti-CD3 VH CDR3 contains the sequence SEQ ID NO: 4; (c) the anti-CD3 VL CDR1 contains the sequence SEQ ID NO: 77, 85, 91, 278, 279 or 280; The anti-CD3 VL CDR2 contains the sequence SEQ ID NO: 78 or 281; and the anti-CD3 VL CDR3 contains the sequence SEQ ID NO: 79; and (d) the VH CDR1 against GUCY2c contains the sequence SEQ ID NO: 12, 20, 27, 34, 42, 74, 257, 258, 259, 260, or 261; The VH CDR2 against GUCY2c contains the sequence SEQ ID NO: 13, 21, 28, 35, 43, 53, 66, 68, 70, 72, 75, 262, 263, 264, 265, 266 or 267; and CDR3 VH against GUCY2c contains the sequence SEQ ID NO: 14, 22, 29, 36 or 44.

In some such embodiments, the present invention provides a bispecific antibody wherein: (a) the anti-GUCY2c VH CDR1 comprises the sequence SEQ ID NO: 74 or 259; (b) CDR2 VH against GUCY2c contains the sequence SEQ ID NO: 75 or 267; (c) CDR3 VH against GUCY2c contains the sequence SEQ ID NO: 29; (d) CDR1 VL against GUCY2c contains the sequence SEQ ID NO: 148; (e) CDR2 VL against GUCY2c contains the sequence SEQ ID NO: 149; and (f) the VL anti-GUCY2c CDR3 contains the sequence of SEQ ID NO: 142.

In further embodiments as described above, the invention provides a bispecific antibody wherein: (a) the anti-CD3 VH CDR1 comprises the sequence SEQ ID NO: 2, 268, or 277; (b) CDR2 VH against CD3 contains the sequence SEQ ID NO: 10 or 270; (c) The anti-CD3 VH CDR3 contains the sequence SEQ ID NO: 4; (d) the anti-CD3 VL CDR1 contains the sequence SEQ ID NO: 91, 278, 279 or 280; (e) VL anti-CD3 CDR2 contains the sequence SEQ ID NO: 78 or 281; and (f) the anti-CD3 VL CDR3 contains the sequence SEQ ID NO: 79.

In a specific embodiment, the present invention provides a polynucleotide comprising a nucleotide sequence encoding the variable regions of an antibody heavy chain and/or light chain as set forth herein and in Table 5, Table 37A and Table 37B, including: CD3-0001, CD3- 0004, CD3-0006, GUCY2C-0074, GUCY2C-0077, GUCY2C-0078, GUCY2C-0098, GUCY2C-0104, GUCY2C-0105, GUCY2C-0240, GUCY2C-0315, GUCY2C-0179, GUCY2C-0193, GUCY2C-0210, GUCY2C-0212, GUCY2C-0241, GUCY2C-0247, GUCY2C-1186, GUCY2C-1467, GUCY2C-1478, GUCY2C-1481, GUCY2C-1512, GUCY2C-1518, GUCY2C-1526, GUCY2C-1527, GUCY 2C-1538, GUCY2C- 1554, GUCY2C-1555, GUCY2C-1556, GUCY2C-1557, GUCY2C-1590, GUCY2C-1591, GUCY2C-1592, GUCY2C-1608, huIGHV3-7, huIGKV1-39, GUCY2C-0405, GUCY2C-0486, GUCY2C-1640, GUCY2C-0250, GUCY2C-1678, GUCY2C-1679 or GUCY2C-1680.

The sequences presented herein include anti-GUCY2c antibodies, anti-CD3 antibodies and anti-CD3-GUCY2c bispecific antibodies. In some embodiments, these antibodies are designated by their clone ID (eg, GUCY2C-0098). These clone IDs can be used to designate the mature antibody (eg, IgG or anti-CD3 bispecific antibody). The clone ID may also refer to the unique VH or VL regions found in the antibody. In the case of clone IDs corresponding to anti-CD3-GUCY2c bispecific antibodies (eg, GUCY2C-0098), a unique region of the anti-CD3 antibody (eg, VL and/or VH regions) included in the mature protein may also be mentioned. Table 5 shows the format of the antibody and the anti-CD3 region (if present) present in the mature protein. Tables 37A and 37B further provide the sequences of the components of these mature anti-GUCY2c antibodies, anti-CD3 IgG and anti-CD3-GUCY2c bispecific antibodies.

Table 5

Subsequence Description Format Anti-CD3 region CD3-0001 Anti-CD3 antibody IgG CD3-0001 CD3-0004 Anti-CD3 antibody IgG CD3-0004 CD3-0006 Anti-CD3 antibody IgG CD3-0006 GUCY2C-0074 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0077 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0078 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0098 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0104 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0105 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0240 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0315 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0179 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-0193 Antibody against GUCY2c IgG No GUCY2C-0210 Antibody against GUCY2c IgG No GUCY2C-0212 Antibody against GUCY2c IgG No GUCY2C-0241 Antibody against GUCY2c IgG No GUCY2C-0247 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0004 GUCY2C-1186 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1467 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1476 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1478 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1481 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1512 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1518 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1526 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1527 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1538 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1554 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1555 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1556 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1557 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1590 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1591 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1592 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 GUCY2C-1608 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0006 huIGHV3-7 Human germline immunoglobulin VH VH But huIGKV1-39 Human germline immunoglobulin VL VL But GUCY2C-0405 scFv-Fc fusion protein against GUCY2c scFv-Fc fusion protein But GUCY2C-0486 Antibody against GUCY2c IgG But GUCY2C-1640 Antibody against GUCY2c IgG But GUCY2C-0250 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody CD3-0001 GUCY2C-1678 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody 2B5-1038 GUCY2C-1679 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody 2B5-1039 GUCY2C-1680 Bispecific antibody against CD3-GUCY2c Bispecific anti-CD3 antibody 2B5-1040

Some of the anti-CD3 antibodies presented herein can be referred to by several names. For example, CD3-0001 can be referred to as 2B5v1 or h2B5v1; CD3-0004 may be referred to as 2B4 or h2B4; and CD3-0006 may be referred to as 2B5v6 or h2B5v6.

Some of the anti-CD3 antibodies provided herein include, but are not limited to, CD3-0006 variants (also referred to as 2B5v6 or h2B5v6). For example, variants of CD3-0006 include, but are not limited to, 2B5-1038, 2B5-1039, and 2B5-1040.

The binding affinity (K _D ) of an antibody provided herein to GUCY2c (such as human GUCY2c (e.g., SEQ ID NO: 224, 230, or 232)) or CD3 (SEQ ID NO: 242) can range from about 0.001 nM to approximately 6500 nM. In some embodiments, the binding affinity is approximately 6500 nM, 6000 nM, 5500 nM, 4500 nM, 4000 nM, 3500 nM, 3000 nM, 2500 nM, 2000 nM, 1500 nM, 1000 nM, 750 nM, 500 nM, 400 nM, 300 nM, 250 nM, 200 nM, 150 nM, 100 nM, 75 nM, 50 nM, 45 nM, 40 nM, 35 nM, 30 nM, 25 nM, 20 nM, 19 nM, 17 nM, 16 nM, 15 nM , 10 nM, 8 nM, 7.5 nM, 7 nM, 6.5 nM, 6 nM, 5.5 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, 0.5 nM, 0, 3 nM, 0.1 nM, 0.01 nM, 0.002 nM or 0.001 nM. In some embodiments, the binding affinity is less than about 6500 nM, 6000 nM, 5500 nM, 5000 nM, 4000 nM, 3000 nM, 2000 nM, 1000 nM, 900 nM, 800 nM, 500 nM, 250 nM, 200 nM, 100 nM , 50 nM, 30 nM, 20 nM, 10 nM, 7.5 nM, 7 nM, 6.5 nM, 6 nM, 5 nM, 4.5 nM, 4 nM, 3.5 nM, 3 nM, 2, 5 nM, 2 nM, 1.5 nM, 1 nM, 0.5 nM, 0.1 nM, 0.05 nM, 0.01 nM or less. In some embodiments, the antibody has a K _D of from about 80 to about 200 pM, preferably from about 100 to about 150 pM.

In some embodiments, the present invention provides an antibody that binds to GUCY2c and/or CD3 and competes with an antibody provided herein, including: CD3-0001, CD3-0004, CD3-0006, GUCY2C-0074, GUCY2C-0077, GUCY2C -0078, GUCY2C-0098, GUCY2C-0104, GUCY2C-0105, GUCY2C-0240, GUCY2C-0315, GUCY2C-0179, GUCY2C-0193, GUCY2C-0210, GUCY2C-0212, GUCY2C-0241, GUCY2C-02 47, GUCY2C-1186 , GUCY2C-1467, GUCY2C-1478, GUCY2C-1481, GUCY2C-1512, GUCY2C-1518, GUCY2C-1526, GUCY2C-1527, GUCY2C-1538, GUCY2C-1554, GUCY2C-1555, GUCY2C-1556, G UCY2C-1557, GUCY2C -1590, GUCY2C-1591, GUCY2C-1592, GUCY2C-1608, huIGHV3-7, huIGKV1-39, GUCY2C-0405, GUCY2C-0486, GUCY2C-1640, GUCY2C-0250, GUCY2C-1678, GUCY2C-1679 and GUCY2C-1680 (Table 5).

In one aspect, the present invention provides an antibody having any of the framework sequences set forth in Table 6. In some embodiments, the present invention provides an antibody as described herein further comprising a human or humanized VH framework and a human or humanized VL framework . In some such embodiments, the VH framework comprises the sequence SEQ ID NO: 5, 6, 7, 8, 15, 16, 17, 18, 23, 24, 25, 30, 31, 32, 37, 38, 39, 40, 45 , 46, 47, 49, 50, 51, 54, 55, 56, 58, 59, 61 or 63; and/or the VL framework contains the sequence SEQ ID NO: 80, 81, 82, 83, 86, 87, 88, 89, 96, 97, 98, 99, 103, 110, 111, 116, 117, 118, 122, 123 , 124, 126, 127, 128, 130, 131, 132, 133, 135, 139 or 155.

Table 6

Description of the wireframe area Subsequence CD3-0001_VH FW_H1 EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO: 5) CD3-0001_VH FW_H2 WVRQAPGKGLEWVA (SEQ ID NO: 6) CD3-0001_VH FW_H3 RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 7) CD3-0001_VH FW_H4 WGQGTTVTVSS (SEQ ID NO: 8) GUCY2C-0074_VH FW_H1 EVQLQQSGAELARPGASVNLSCKAS (SEQ ID NO: 15) GUCY2C-0074_VH FW_H2 WVKQRPGQGLEWIG (SEQ ID NO: 16) GUCY2C-0074_VH FW_H3 KATLTADKSSSTAYMQLSSLASEDSAVYYCVR (SEQ ID NO: 17) GUCY2C-0074_VH FW_H4 WGQGTSVTVSS (SEQ ID NO: 18) GUCY2C-0077_VH FW_H1 EVQLQQSGAELARPGASVKLSCKAS (SEQ ID NO: 23) GUCY2C-0077_VH FW_H2 WIKQRPGQGLEWIG (SEQ ID NO: 24) GUCY2C-0077_VH FW_H3 KATLTADKSSNTAYMQLSSLASEDSAVYYCAR (SEQ ID NO: 25) GUCY2C-0098_VH FW_H1 QVQLQQPGAELVKPGASVKLSCKAS (SEQ ID NO: 30) GUCY2C-0098_VH FW_H3 KATLTVDKSATTAYMQLSSLTAEDSAVYYCTR (SEQ ID NO: 31) GUCY2C-0098_VH FW_H4 WGAGTTVTVSS (SEQ ID NO: 32) GUCY2C-0104_VH FW_H1 EVQLQQSGAELVKPGASVKLSCTAS (SEQ ID NO: 37) GUCY2C-0104_VH FW_H2 WVKQRPEQGLEWIG (SEQ ID NO: 38) GUCY2C-0104_VH FW_H3 KATITADTSSNTAYLQLSSLTSEDTAVFYCSS (SEQ ID NO: 39) GUCY2C-0104_VH FW_H4 WGQGTTLTVSS (SEQ ID NO: 40) GUCY2C-0105_VH FW_H1 EVQLQQSGPELVKPGASVKISCKAS (SEQ ID NO: 45) GUCY2C-0105_VH FW_H2 WVKQSHGKSLEWIG (SEQ ID NO: 46) GUCY2C-0105_VH FW_H3 KATLTVDKSSSTAYMHLNSLTSEDSAVYYCAR (SEQ ID NO: 47) GUCY2C-0240_VH FW_H2 WVRQAPGKGLEWIG (SEQ ID NO: 49) GUCY2C-0240_VH FW_H3 RFTISADKAKNSAYLQMNSLRAEDTAVYYCVR (SEQ ID NO: 50) GUCY2C-0240_VH FW_H4 WGQGTLVTVSS (SEQ ID NO: 51) GUCY2C-0315_VH FW_H1 QVQLVQSGAEVKKPGASVKVSCKAS (SEQ ID NO: 54) GUCY2C-0315_VH FW_H2 WVRQAPGQGLEWIG (SEQ ID NO: 55) GUCY2C-0315_VH FW_H3 RVTMTRDTSTSTVYMELSSLRSEDTAVYYCVR (SEQ ID NO: 56) GUCY2C-0179_VH FW_H3 RFTISVDKAKNSAYLQMNSLRAEDTAVYYCAR (SEQ ID NO: 58) GUCY2C-0179_VH FW_H4 WGQGTMVTVSS (SEQ ID NO: 59) GUCY2C-0193_VH FW_H3 RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR (SEQ ID NO: 61) GUCY2C-0210_VH FW_H2 WVRQAPGKGLEWIA (SEQ ID NO: 63) CD3-0001_VL FW_L1 DIQMTQSPSSLSSASVGDRVTITC (SEQ ID NO: 80) CD3-0001_VL FW_L2 WYQQKPGKAPKLLIY (SEQ ID NO: 81) CD3-0001_VL FW_L3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO: 82) CD3-0001_VL FW_L4 FGGGTKVEIK (SEQ ID NO: 83) CD3-0004_VL FW_L1 DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 86) CD3-0004_VL FW_L2 WYQQKPGQPPKLLIS (SEQ ID NO: 87) CD3-0004_VL FW_L3 GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 88) CD3-0004_VL FW_L4 FGSGTKLEIK (SEQ ID NO: 89) GUCY2C-0074_VL FW_L1 DIVLTQSPASLAVSLGQRATISC (SEQ ID NO: 96) GUCY2C-0074_VL FW_L2 WFQQKPGQPPKLLIY (SEQ ID NO: 97) GUCY2C-0074_VL FW_L3 GVPARFSGSGSGTDFSLNIHPLEEDDTAMFFC (SEQ ID NO: 98) GUCY2C-0074_VL FW_L4 FGGGTKLEIK (SEQ ID NO: 99) GUCY2C-0077_VL FW_L3 GVPARFSGSGSGTDFSLNIHPMEEDDTAMYFC (SEQ ID NO: 103) GUCY2C-0098_VL FW_L2 WYQQKPGQPPKLLIY (SEQ ID NO: 110) GUCY2C-0098_VL FW_L3 GVPARFSGSGSGTDFSLNIHPVEEDDIAMYFC (SEQ ID NO: 111) GUCY2C-0104_VL FW_L1 DIVMTQSPASLAVSLGQRATISC (SEQ ID NO: 116) GUCY2C-0104_VL FW_L3 GVPARFSGSGSGTDFTLNIHPVEEEDAATYYC (SEQ ID NO: 117) GUCY2C-0104_VL FW_L4 FGAGTKLELK (SEQ ID NO: 118) GUCY2C-0105_VL FW_L1 DIVMTQSPSSLAVSVGEKVTVSC (SEQ ID NO: 122) GUCY2C-0105_VL FW_L2 WYQQRPGQSPKLLIY (SEQ ID NO: 123) GUCY2C-0105_VL FW_L3 GVPDRFTGSGSGTDFTLSISSVKAEDLAVYYC (SEQ ID NO: 124) GUCY2C-0240_VL FW_L1 DIQLTQSPSSLSSASVGDRVTITC (SEQ ID NO: 126) GUCY2C-0240_VL FW_L2 WFQQKPGKAPKLLIY (SEQ ID NO: 127) GUCY2C-0240_VL FW_L4 FGQGTKLEIK (SEQ ID NO: 128) GUCY2C-0315_VL FW_L1 EIVLTQSPATLSLSPGERATLSC (SEQ ID NO: 130) GUCY2C-0315_VL FW_L2 WYQQKPGQAPRLLIY (SEQ ID NO: 131) GUCY2C-0315_VL FW_L3 GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC (SEQ ID NO: 132) GUCY2C-0315_VL FW_L4 FGQGTKVEIK (SEQ ID NO: 133) GUCY2C-0179_VL FW_L2 WYQQKPGKSPKLLIY (SEQ ID NO: 135) GUCY2C-0247_VL FW_L2 WYQQKPGKPPKLLIYm (SEQ ID NO: 139) GUCY2C-1512_VL FW_L4 FGQGTKLKIK (SEQ ID NO: 155)

In one embodiment, the present invention provides an antibody wherein the VH region comprises the sequence SEQ ID NO: 73 or a variant thereof with one or more conservative amino acid substitutions at residues not located in the CDR region; and/or wherein the VL region contains the sequence SEQ ID NO: 147 or a variant thereof with one or more conservative amino acid substitutions in amino acids not found in the CDR region.

In one aspect, the present invention provides a bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain. In one embodiment, the first polypeptide chain comprises an amino acid sequence set forth in one or more of SEQ ID NOs: 216 and 220. In another embodiment, the second polypeptide chain comprises an amino acid sequence set forth in one or more of SEQ ID NOs: 216 and 220 In a preferred embodiment, the first polypeptide chain comprises the amino acid sequence SEQ ID NO: 216; and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 220. In some such embodiments, the bispecific antibody may specifically bind to a GUCY2c epitope and a CD3 epitope and comprises a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises the sequence of SEQ ID NO: 216 and the second polypeptide chain comprises the sequence SEQ ID NO: 220. In one embodiment, the second polypeptide chain is any polypeptide subject to binding to the first polypeptide chain through a contact region.

In one such embodiment, the present invention provides a bispecific antibody wherein: (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-GUCY2c antibody VL (anti-GUCY2c VL) and VH anti-CD3 antibodies (VH anti-CD3), and (ii) the first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in a direction from N-terminus to C-terminus: (i) domain 2 containing anti-CD3 VL (anti-CD3 VL) and anti-GUCY2c VH (anti-GUCY2c VH), and (ii) a second heterodimerization promoting domain; where VL against GUCY2c and VH against GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

In another such embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-CD3 VL and anti-GUCY2c VH, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-GUCY2c VL and anti-CD3 VH, and (ii) a second heterodimerization promoting domain; where VL against GUCY2c and VH against GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

In another such embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-GUCY2c VH and anti-CD3 VL, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-CD3 VH and anti-GUCY2c VL, and (ii) a second heterodimerization promoting domain; where VH against GUCY2c and VL against GUCY2c form a domain that specifically binds to GUCY2c; and VH anti-CD3 and VL anti-CD3 form a domain that specifically binds to CD3.

In a further embodiment, (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-CD3 VH and anti-GUCY2c VL, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain comprises, in a direction from N-terminus to C-terminus: (i) domain 2 containing anti-GUCY2c VH and anti-CD3 VL, and (ii) a second heterodimerization promoting domain, wherein anti-GUCY2c VL and anti-CD3 VH GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

The invention also relates to isolated polynucleotides encoding antibodies of the invention, and vectors and host cells containing the polynucleotides. In one aspect, the polynucleotide comprises a nucleotide sequence encoding heavy chain variable regions, light chain variable regions, CDR regions, framework regions, peak and valley Fc chains, first polypeptide chains, and second polypeptide chains set forth in Tables 1-7 and 38 In some embodiments, the polynucleotide comprises a sequence encoding any of the antibodies CD3-0001, CD3-0004, CD3-0006, GUCY2C-0074, GUCY2C-0077, GUCY2C-0078, GUCY2C-0098, GUCY2C-0104, GUCY2C-0105, GUCY2C -0240, GUCY2C-0315, GUCY2C-0179, GUCY2C-0193, GUCY2C-0210, GUCY2C-0212, GUCY2C-0241, GUCY2C-0247, GUCY2C-1186, GUCY2C-1467, GUCY2C-1478, GUCY2C-14 81, GUCY2C-1512 , GUCY2C-1518, GUCY2C-1526, GUCY2C-1527, GUCY2C-1538, GUCY2C-1554, GUCY2C-1555, GUCY2C-1556, GUCY2C-1557, GUCY2C-1590, GUCY2C-1591, GUCY2C-1592, G UCY2C-1608, huIGHV3 -7, huIGKV1-39, GUCY2C-0405, GUCY2C-0486, GUCY2C-1640, GUCY2C-0250, GUCY2C-1678, GUCY2C-1679 or GUCY2C-1680.

The coding sequence for the antibody of interest can be maintained in a vector in a host cell, and the host cell can then be grown and frozen for later use. Vectors (including expression vectors) and host cells are further provided herein.

The invention also relates to fusion proteins containing one or more fragments or regions from the antibodies of the present invention. In one embodiment, the present invention provides a fusion polypeptide comprising at least 10 contiguous amino acids from the light chain variable region set forth in SEQ ID NO: 76, 84, 90, 92, 100, 104, 106, 112, 119, 125 , 129, 134, 136, 137, 138, 140, 143, 145, 147, 150, 152, 156, 158, 160, 162, 166, 170, 171, 172, 173, 174, 175, 274, 275 or 2 76 ; and/or at least 10 contiguous amino acids from the heavy chain variable region set forth in SEQ ID NO: 1, 9, 11, 19, 26, 33, 41, 48, 52, 57, 60, 62, 64, 65, 67 , 69, 71, 73, or 273. In other embodiments, the present invention provides a fusion polypeptide comprising at least about 10, at least about 15, at least about 20, at least about 25, or at least about 30 contiguous light chain variable region amino acids and/or at least about 10, at least about 15, at least about 20, at least about 25, or at least about 30 contiguous heavy chain variable region amino acids. In another embodiment, the fusion polypeptide comprises a light chain variable region and/or a heavy chain variable region shown in any of the sequence pairs selected from SEQ ID NOs: 73 and 147; 69 and 147; 60 and 138; 48 and 125; or 26 and 106. In another embodiment, the fusion polypeptide contains one or more CDRs. In other embodiments, the fusion polypeptide comprises CDR H3 (CDR3 VH) and/or CDR L3 (CDR3 VL). For purposes of the present invention, the fusion protein contains one or more antibodies and another amino acid sequence to which it is not attached in the native molecule, for example, a heterologous sequence or a homologous sequence from another region. Non-limiting examples of heterologous sequences include a tag such as a FLAG tag or a 6His tag. The marks are well known in this field. The fusion polypeptide can be produced by methods known in the art, for example, synthetically or recombinantly. Typically, the fusion proteins of the present invention are produced by producing the expression polynucleotide encoding them by recombinant methods as described herein, although they can also be produced by other methods known in the art, including, for example, chemical synthesis.

The invention also relates to the scFv antibodies of the present invention. Single chain variable region fragments are produced by linking light and/or heavy chain variable regions using a short connecting peptide (Bird et al., Science 242:423-426, 1988). An example of a connecting peptide is GGGGSGGGGSGGGGSG (SEQ ID NO: 193), which serves as an approximately 3.5 nm bridge between the carboxy terminus of one variable region and the amino terminus of another variable region. Linkers with other sequences can be designed and used (Bird et al., Science 242:423-426, 1988). Linkers should be short, flexible polypeptides and preferably contain less than about 20 amino acid residues. In turn, linkers can be modified for additional functions, such as attaching drugs or attaching to solid supports. Single-chain variants can be produced recombinantly or synthetically. An automatic synthesizer can be used to synthesize scFv. To recombinantly produce a scFv, a suitable plasmid containing a polynucleotide encoding the scFv can be inserted into a suitable host cell, eukaryotic such as yeast cells, plant cells, insect cells or mammalian cells, or prokaryotic such as E. coli . Polynucleotides encoding the scFv of interest can be prepared by conventional methods, such as polynucleotide ligation. The resulting scFv can be isolated using standard protein purification methods known in the art.

Other forms of single chain antibodies are also provided, such as diabodies or minibodies. Diabodies are bivalent, bispecific antibodies in which the heavy chain variable regions (VH) and light chain variable regions (VL) are expressed on a single polypeptide chain, but using a linker too short to pair between the two domains on the same chain, thus , causes the domains to pair with complementary domains of the other chain and results in the formation of two antigen-binding sites (see, for example, Holliger, P., et al., Proc. Natl. Acad Sci. USA 90:6444-6448, 1993; Poljak, R. J., et al., Structure 2:1121-1123, 1994). The minibody comprises the VL and VH regions of the native antibody fused to the hinge region and CH3 domain of the immunoglobulin molecule. See, for example, US Pat. No. 5,837,821.

In one aspect, the present invention relates to compositions (such as pharmaceutical compositions) containing any of the polynucleotides of the invention. In some embodiments, the composition comprises an expression vector containing a polynucleotide encoding any of the antibodies provided herein. In other embodiments, the composition comprises either or both of the polynucleotides set forth in SEQ ID NO: 246 (encoding the first polypeptide chain of GUCY2C-1608) and SEQ ID NO: 247 (encoding the second polypeptide chain of GUCY2C-1608). Expression vectors and administration of polynucleotide compositions are presented hereinafter.

Polynucleotides complementary to any such sequences are also included in the present invention. Polynucleotides can be single-stranded (coding or antisense) or double-stranded and can be DNA (genomic, cDNA or synthetic) or RNA molecules. RNA molecules include mature and immature mRNA, such as precursor mRNA (pre-mRNA) or heterogeneous nuclear mRNA (hnRNA) and mature mRNA. Additional coding or non-coding sequences may optionally be present in the polynucleotide of the present invention, and the polynucleotide may optionally be linked to other molecules and/or support materials.

Polynucleotides may contain a native sequence (ie, an endogenous sequence encoding an antibody or part thereof) or a variant of such a sequence. Polynucleotide variants contain one or more substitutions, additions, deletions and/or insertions such that the immunoreactivity of the encoded polypeptide is not reduced relative to the native immunoreactive molecule. The effect on the immunoreactivity of the encoded polypeptide can generally be assessed as presented herein. Preferably, the variants exhibit at least about 70% identity, more preferably at least about 80% identity, even more preferably at least about 90% identity, and most preferably at least about 95% identity with respect to polynucleotide sequence , encoding the native antibody or part thereof.

Two polynucleotide or polypeptide sequences are said to be “identical” if the sequence of nucleotides or amino acids in the two sequences is the same when aligned for maximum matching, as described below. Comparison of two sequences is typically accomplished by comparing the sequences in a comparison window to identify and compare local regions of sequence similarity. As used herein, the term "comparison window" refers to a segment of at least about 20 contiguous positions, typically 30 to about 75 or 40 to about 50, in which the sequence can be compared to a reference sequence with the same number of contiguous positions after optimal alignment of two sequences.

Optimal sequence alignment for comparison can be performed using the Megalign program in the Lasergene bioinformatics software package (DNASTAR, Inc., Madison, WI) using default parameters. Preferably, "percentage sequence identity" is determined by comparing two optimally aligned sequences in a comparison window of at least 20 positions, where a portion of the polynucleotide or polypeptide sequence in the comparison window may contain additions or deletions (i.e., omissions) of 20 percent or less, typically 5 to 15 percent or 10 to 12 percent, compared to reference sequences (not containing additions or deletions) for optimal alignment of two sequences. The percentage is calculated by determining the number of positions at which identical nucleic acid bases or amino acid residues occur in both sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the reference sequence (i.e., window size) and multiplying the result by 100 to obtain percent sequence identity.

Variants, additionally or alternatively, may be substantially homologous to the native gene or a portion or complementary sequence thereof. Such polynucleotide variants can hybridize under moderately stringent conditions with the natural DNA sequence encoding the native antibody (or complementary sequence).

Those skilled in the art will appreciate that, as a result of the degeneracy of the genetic code, there are many nucleotide sequences encoding the polypeptide disclosed herein. Some of these polynucleotides have minimal homology to the nucleotide sequence of any native gene. However, polynucleotides varying due to differences in codon usage are specifically included in the present invention. In addition, alleles of genes containing polynucleotide sequences presented in the present description are included in the scope of the present invention. Alleles are endogenous genes that have been altered by one or more mutations, such as deletions, additions and/or nucleotide substitutions. The resulting mRNA and protein may optionally have altered structure or function. Alleles can be identified by standard methods (such as hybridization, amplification and/or comparison of sequences from databases).

In one aspect, the present invention relates to a method for producing any of the polynucleotides presented herein. For example, the polynucleotides of the present invention can be produced using chemical synthesis, recombinant methods or PCR. Methods for chemically synthesizing polynucleotides are well known in the art and do not require detailed discussion herein. One skilled in the art can use the sequences presented herein and a commercial DNA synthesizer to obtain the desired DNA sequence.

To produce polynucleotides by recombinant methods, a polynucleotide containing the desired sequence can be inserted into a suitable vector, and the vector, in turn, can be inserted into a suitable host cell for replication and amplification, as provided herein. Polynucleotides can be incorporated into host cells by any methods known in the art. Cells are transformed by insertion of an exogenous polynucleotide through direct uptake, endocytosis, transfection, F-crossing, or electroporation. Once inserted, the exogenous polynucleotide can be maintained in the cell as a non-integrated vector (such as a plasmid) or integrated into the genome of the host cell. The polynucleotide thus amplified can be isolated from the host cell by methods well known in the art (eg, Sambrook et al., 1989).

Alternatively, PCR allows DNA sequences to be reproduced. PCR technology is well known in the art and is described in US Patent Nos. 4,683,195, 4,800,159, 4,754,065 and 4,683,202, as well as PCR: Polymerase Chain Reaction, Mullis et al. eds., Birkauswer Press, Boston, 1994.

The RNA can be produced using the isolated DNA in a suitable vector and inserted into a suitable host cell. When a cell divides and DNA is transcribed into RNA, the RNA can be isolated by methods well known to those skilled in the art and described, for example, in Sambrook et al., 1989, supra.

Suitable cloning vectors can be constructed by standard methods or can be selected from the large number of cloning vectors available in the art. Although the cloning vector chosen may vary depending on the host cell used, cloning vectors that can be used will generally have the ability to self-replicate, may have a single target for a particular restriction endonuclease, and/or may carry genes for a marker that can be used in the selection of clones containing the vector. Suitable examples include plasmids and bacterial viruses, e.g. pUC18, pUC19, Bluescript (e.g. pBS SK+) and derivatives thereof, mp18, mp19, pBR322, pMB9, ColE1, pCR1, RP4, phage DNA and shuttle vectors such as pSA3 and pAT28 . These and many other cloning vectors are available from commercial sources such as BioRad, Strategene, and Invitrogen.

Expression vectors are typically replicable polynucleotide constructs containing a polynucleotide of the invention. It should be understood that the expression vector must replicate in host cells as episomes or an integral portion of chromosomal DNA. Suitable expression vectors include, but are not limited to, plasmids, viral vectors, including adenoviruses, adeno-associated viruses, retroviruses, cosmids, and expression vectors described in PCT Publication No. WO87/04462. Vector components may generally include, but are not limited to, one or more of the following: a signal sequence; origin of replication; one or more marker genes; suitable transcriptional control elements (such as promoters, enhancers and terminator). Expression (ie, translation) typically also requires one or more translation control elements, such as ribosome binding sites, translation initiation sites, and stop codons.

Vectors containing polynucleotides of interest can be introduced into a host cell by any of a number of suitable methods, including electroporation, transfection using calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran or other substances; microparticle bombardment; lipofection and infection (for example, if the vector is an infectious agent such as vaccinia virus). The choice of vectors or polynucleotides to be inserted will often depend on the characteristics of the host cell.

Any host cell capable of overexpressing heterologous DNA can be used to isolate genes encoding an antibody, polypeptide or protein of interest. Non-limiting examples of mammalian host cells include COS, HeLa and CHO cells. Also see PCT Publication No. WO87/04462. Suitable non-mammalian host cells include prokaryotes (such as E. coli or B. subtillis ) and yeasts (such as S. cerevisae , S. pombe or K. lactis ). Preferably, the host cells express the cDNA at a level approximately 5 times higher, more preferably 10 times higher, even more preferably 20 times higher than the corresponding endogenous antibody or GUCY2c protein or GUCY2c domain (e.g., domains 1 -4), as determined by immunoassay or FACS. A cell overexpressing an antibody or protein of interest can be identified.

In one aspect, the anti-GUCY2c antibody molecule will have an affinity for GUCY2c, for example, as measured by direct binding assays or competitive binding assays in the picomole to micromolar affinity range, preferably in the picomole to low nanomolar range.

Bispecific antibodies can be produced using the antibodies presented herein. Methods for producing bispecific antibodies are known in the art (see, for example, Suresh et al., Methods in Enzymology 121:210, 1986). Conventionally, recombinant production of bispecific antibodies is based on the coexpression of two immunoglobulin heavy chain-light chain pairs, with the two heavy chains having different specificities (Millstein and Cuello, Nature 305, 537-539, 1983).

In another embodiment, a bispecific antibody provided herein comprises a full-length human antibody, wherein the heterodimeric protein antibody variable region can recruit the activity of a human immune effector cell by specifically binding to an effector antigen (e.g., CD3 antigen) located on the human immune effector cell and wherein the second antibody variable region of the heterodimeric protein can specifically bind to a target antigen. In some embodiments, the human antibody is of an IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the heterodimeric protein comprises an immunologically inert Fc chain.

The human immune effector cell may be any of a variety of immune effector cells known in the art. For example, the immune effector cell may be a member of a human lymphoid cell lineage, including, but not limited to, a T cell (eg, a cytotoxic T cell), a B cell, and a natural killer (NK) cell. The immune effector cell may also be a member of the human myeloid lineage, including, but not limited to, a monocyte, a neutrophil granulocyte, and a dendritic cell. Such immune effector cells may have a cytotoxic or apoptotic effect on a target cell or other desired effect upon activation by binding of the effector antigen.

An effector antigen is an antigen (eg, a protein or polypeptide) expressed on a human immune effector cell. Non-limiting examples of effector antigens that can bind to a heterodimeric protein (eg, a heterodimeric antibody or a bispecific antibody) include human CD3 (or CD3 complex (cluster of differentiation)), CD16, NKG2D, NKp46, CD2, CD28, CD25, CD64, and CD89.

The target cell may be a cell that is native or foreign to humans. In the case of a native target cell, the cell may transform into a malignant cell or become pathologically modified (eg, a native target cell infected with a virus, plasmodium, or bacterium). In the case of a foreign target cell, the cell is an invading pathogen such as a bacterium, plasmodium, or virus.

The target antigen is expressed on a target cell in a disease state (e.g., inflammatory disease, proliferative disease (e.g., cancer), immunological disorder, neurological disease, neurodegenerative disease, autoimmune disease, infectious disease (e.g., viral infection or parasitic infection), allergic reaction, graft-versus-host disease, or host-versus-graft disease). The target antigen is not an effector antigen. Non-limiting examples of target antigens include GUCY2c, BCMA, EpCAM (epithelial cell adhesion molecule), CCR5 (chemokine receptor type 5), CD19, HER (human epidermal growth factor receptor)-2/neu, HER-3, HER-4, EGFR (epidermal growth factor receptor), PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC7, CIhCG, Lewis antigen Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl -GD3, GM2, Globo H, fucosyl-GM1, poly-SA, GD2, carbonic anhydrase IX (MN/CA IX), CD44v6, Shh (Sonic Hedgehog), Wue-1, plasma cell antigen, (membrane-bound) IgE, MCSP ( melanoma chondroitin sulfate proteoglycan), CCR8, TNF-alpha precursor, STEAP, mesothelin, A33 antigen, PSCA (prostate stem cell antigen), Ly-6; desmoglein 4, neoepitope E-cadherin, fetal acetylcholine receptor, CD25, CA19-9 marker, CA-125 marker and MIS (Mullerian inhibitory substance) type II receptor, sTn (sialylated Tn antigen; TAG-72), FAP (fibroblast activation antigen ), endosialin, EGFRvIII, LG, SAS and CD63.

In one embodiment, the present invention provides a bispecific antibody, wherein: (a) the anti-GUCY2c VL CDR1, anti-GUCY2c VL CDR2, and anti-GUCY2c VL CDR3 of the anti-GUCY2c VL contain the sequence set forth in SEQ ID NO: 92, 100, 104, 106, 112, 119, 125, 129, 134, 136, 137, 138, 140, 143, 145, 147, 150, 152, 156, 158, 160, 162, 166, 170, 171, 172, 173, 174 or 175; (b) CDR1 VH against CD3, CDR2 VH against CD3 and CDR3 VH against CD3 of VH against CD3 contain the sequence shown in SEQ ID NO: 1, 9 or 273; (c) CDR1 VL anti-CD3, CDR2 VL anti-CD3 and CDR3 VL anti-CD3 of VL anti-CD3 contain the sequence shown in SEQ ID NO: 76, 84, 90 or 274, 275 or 276; and/or (d) CDR1 VH against GUCY2c, CDR2 VH against GUCY2c and CDR3 VH against GUCY2c from VH against GUCY2c contain the sequence shown in SEQ ID NO: 11, 19, 26, 33, 41, 48, 52, 57, 60 , 62, 64, 65, 67, 69, 71 or 73.

In a specific embodiment, the present invention provides a bispecific antibody wherein: (a) the anti-GUCY2c VH region comprises the sequence SEQ ID NO: 73; and (b) the VL region against GUCY2c contains the sequence SEQ ID NO: 147.

In some embodiments, the present invention provides a bispecific antibody wherein: (a) the anti-GUCY2c VL CDR1 comprises the sequence SEQ ID NO: 93, 101, 105, 107, 113, 120, 148, 153, 163, or 167; CDR2 VL against GUCY2c contains the sequence SEQ ID NO: 78, 94, 102, 108, 114, 141, 144, 146, 149, 151, 157, 159, 161, 164 or 168; and CDR3 VL against GUCY2c contains the sequence SEQ ID NO: 95, 109, 115, 121, 142, 154, 165 or 169; (b) the anti-CD3 VH CDR1 contains the sequence SEQ ID NO: 2, 268 or 277; The anti-CD3 VH CDR2 contains the sequence SEQ ID NO: 3, 10, 269 or 270; and the anti-CD3 VH CDR3 contains the sequence SEQ ID NO: 4; (c) the anti-CD3 VL CDR1 contains the sequence SEQ ID NO: 77, 85, 91, 278, 279 or 280; The anti-CD3 VL CDR2 contains the sequence SEQ ID NO: 78 or 281; and the anti-CD3 VL CDR3 contains the sequence SEQ ID NO: 79; and (d) the VH CDR1 against GUCY2c contains the sequence SEQ ID NO: 12, 20, 27, 34, 42, 74, 257, 258, 259, 260, or 261; The VH CDR2 against GUCY2c contains the sequence SEQ ID NO: 13, 21, 28, 35, 43, 53, 66, 68, 70, 72, 75, 262, 263, 264, 265, 266 or 267; and CDR3 VH against GUCY2c contains the sequence SEQ ID NO: 14, 22, 29, 36 or 44.

In some embodiments, antibodies that can be used in the present invention are monoclonal antibodies, polyclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab') ₂ , Fv, Fc, etc.), chimeric antibodies, bispecific antibodies, heteroconjugated antibodies, single-chain antibodies (ScFv), mutants thereof, fusion proteins containing an antibody moiety (e.g., domain antibody), humanized antibodies, and any other modified configuration of an immunoglobulin molecule containing an antigen recognition site with the required specificity, including antibody glycosylation variants , variants of the amino acid sequence of antibodies and covalently modified antibodies. The antibodies may be mouse, rat, human, or any other origin (including chimeric or humanized antibodies).

In some embodiments, an anti-GUCY2c or anti-CD3 antibody provided herein is a monoclonal antibody. For example, an antibody against GUCY2c or CD3 is a humanized monoclonal antibody or a chimeric monoclonal antibody.

The present invention relates to a bispecific antibody containing an Fc chain or β domain or a portion thereof. In some embodiments, the Fc chain or portions thereof comprise one or more IgG1, IgG2, IgG3, or IgG4 Fc chain constant domains (eg, a CH2 or CH3 domain). In another embodiment, the present invention relates to molecules containing an Fc chain or a portion thereof, wherein the Fc chain or portion thereof contains at least one amino acid modification (eg, substitution) relative to a comparable wild-type Fc chain or portion thereof. Fc region variants are well known in the art and are primarily used to alter the phenotype of an antibody containing the Fc region variant, as assessed by any binding activity or effector function assay well known in the art, such as an ELISA assay. SPR or ADCC. Such Fc chain variants or portions thereof may enhance the plasma half-life and stability exhibited by a bispecific antibody of the invention containing an Fc chain or portion thereof. In another embodiment, the present invention relates to the use of any Fc variant known in the art.

In one embodiment, one or more Fc chain amino acid modifications are made to reduce the affinity and avidity of the Fc chain and thus the bispecific antibody molecule of the invention to one or more FcγR receptors. In a specific embodiment, the present invention provides bispecific antibodies comprising an Fc chain variant or a portion thereof, wherein the Fc chain variant contains at least one amino acid modification relative to the wild type Fc chain, wherein the Fc region variant binds to only one FcγR, where FcγR is FcγRIIIA. In another specific embodiment, the present invention provides bispecific antibodies comprising an Fc chain variant or a portion thereof, wherein the Fc chain variant contains at least one amino acid modification relative to the wild type Fc chain, wherein the Fc region variant binds to only one FcγR , where FcγR is FcγRIIA. In another specific embodiment, the present invention provides bispecific antibodies comprising an Fc chain variant or a portion thereof, wherein the Fc chain variant contains at least one amino acid modification relative to the wild type Fc chain, wherein the Fc chain variant binds to only one FcγR , where FcγR is FcγRIIB. In another embodiment, the present invention provides molecules containing an Fc chain variant, wherein the variant confers or mediates reduced ADCC activity (or other effector function) and/or increased binding to FcγRIIB (CD32B) relative to a non-Fc chain-containing or Fc-containing molecule -wild-type chain, as measured by methods known to those skilled in the art and described herein.

The invention also relates to the use of an Fc region containing domains or regions of two or more IgG isotypes. As is known in the art, amino acid modification of the Fc region can significantly affect Fc-mediated effector function and/or binding activity. However, these changes in functional characteristics can be further improved and/or modified when implemented in the context of selected IgG isotypes. Likewise, the native characteristics of the Fc isotype can be affected by one or more amino acid modifications. The multiple IgG isotypes (i.e., IgG1, IgG2, IgG3, and IgG4) exhibit different physical and functional properties, including serum half-life, complement fixation, FcγR binding affinities, and activity of effector functions (e.g., ADCC, CDC) due to differences amino acid sequences of hinge regions and/or Fc regions.

In one embodiment, the amino acid modification and Fc regions of the IgG are independently selected based on their respective individual binding function and/or effector function activities to construct a bispecific antibody with the desired characteristics. In a specific embodiment, amino acid modifications and hinge regions/Fc regions of IgG are separately assayed for binding and/or effector function activity as described herein or known in the art for IgG1. In one embodiment, the amino acid modification and the hinge region/Fc region of an IgG exhibit similar functionality, e.g., reduced ADCC activity (or other effector function) and/or increased binding to FcγRIIB in the context of a bispecific antibody or other Fc-containing molecule (e.g. immunoglobulin). In another embodiment, the present invention provides Fc region variants containing combinations of amino acid modifications known in the art and selected IgG regions exhibiting novel properties not detectable when the modifications and/or regions are analyzed independently as provided herein.

In some embodiments, the first heterodimerization-promoting domain and the second heterodimerization-promoting domain each comprise an Fc region comprising a CH2 domain and a CH3 domain, wherein the amino acid sequence of each of the CH2 domain and/or the CH3 domain contains at least one amino acid modification relative to the Fc -wild type region forming a protrusion or depression.

In some embodiments, the first heterodimerization promoting domain and the second heterodimerization promoting domain each comprise a CH2 domain and a CH3 domain, wherein the amino acid sequence of each of the CH2 domains and/or each of the CH3 domains is modified to regulate heterodimerization and/or stabilize the bispecific antibody.

In some such embodiments, the bispecific antibodies of the present invention comprise a first heterodimerization-promoting domain on a first polypeptide chain and a second heterodimerization-promoting domain on a second polypeptide chain. Collectively, the first and second heterodimerization-promoting domains regulate heterodimerization, and/or stabilize the bispecific antibody (eg, through knob-valley interactions on complementary heterodimerization-promoting domains), and/or serve to stabilize the bispecific antibody.

In some embodiments, the first heterodimerization-promoting domain and the second heterodimerization-promoting domain are not both protrusions or are not both recesses; and/or the first heterodimerization-promoting domain and the second heterodimerization-promoting domain form the Fc region of an IgG immunoglobulin.

In some embodiments, the first heterodimerization promoting domain may comprise an Fc chain having a CH2 and/or CH3 domain modified to contain a knob or a trench. In some such embodiments, the amino acid sequence of the CH2 domain and/or the CH3 domain comprises at least one amino acid modification wherein: (a) the CH3 domain of the first heterodimerization promoting domain forms an overhang; and (b) the CH3 domain of the second heterodimerization promoting domain forms a depression. In another such embodiment, the CH3 domain of the first heterodimerization promoting domain contains mutations Y349C and/or T366W to form an overhang; and the CH3 domain of the second heterodimerization promoting domain contains mutations S354C, T366S, L368A, and/or Y407V to form a cavity (numbered according to the EU index). In some specific embodiments, the mutations result in reduced effector function.

In some embodiments, the first heterodimerization promoting domain may comprise a CH2 and/or CH3 domain modified to contain an overhang containing the sequence of SEQ ID NO: 188, if the second heterodimerization promoting domain comprises a CH2 and/or CH3 domain modified such that it contained a depression. In another embodiment, the first heterodimerization-promoting domain may comprise a trench if the second heterodimerization-promoting domain comprises a CH2 and/or CH3 domain modified to contain a protrusion. In a specific embodiment described above, the first heterodimerization promoting domain contains the sequence SEQ ID NO: 188 to form an overhang; and wherein the second heterodimerization promoting domain contains the sequence of SEQ ID NO: 189 to form a cavity. Table 7 shows the amino acid sequences of the Fc chains with peaks and valleys and the first polypeptide chains and second polypeptide chains of bispecific antibodies against GUCY2c with various antibodies against GUCY2c and CD3 presented herein (SEQ ID NO: 196, 197, 199, 200, 202, 203, 205, 206, 210, 211, 216, 217, 219, 220, 248, 249, 282, 283, 284, 285, 286 and 287).

Table 7

Fc Raised Chain (SEQ ID NO: 188) APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPG Cavity Fc Chain (SEQ ID NO: 189) APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0074_protrusion
(SEQ ID NO: 196) DIVLTQSPASLAVSLGQRATISCRASESVDNYGISFMNWFQQKPGQPPKLLIYAASNPGSGVPARFSGSGSGTDFSLNIHPLEEDDTAMFFCQQSKEVPYTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0074_cavity
(SEQ ID NO: 197) DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLQQSGAELARPGASVNLSCKASGYTFTTYWMQWVKQRPGQGLEWIGAIYPGDGMTTYTQKFKDKATLTAD KSSSTAYMQLSSLASEDSAVYYCVRKGMDYWGQGTSVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0098_protrusion
(SEQ ID NO: 199) DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQ MNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0098_cavity
(SEQ ID NO: 200) DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGQVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTV DKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSL SCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0105_protrusion
(SEQ ID NO: 202) DIVMTQSPSSLAVSVGEKVTVSCKSSQSLLYSSNQKNYLAWYQQRPGQSPKLLIYWASTRESGVPDRFTGSGSGTDFTLSISSVKAEDLAVYYCQQYYSYPTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0105_cavity
(SEQ ID NO: 203) DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLQQSGPELVKPGASVKISCKASGYSFTDYIMLWVKQSHGKSLEWIGNSNPYYGSTSYNLKFKGKATLTVDK SSSTAYMHLNSLTSEDSAVYYCARSGYYGSSPYWYFDVWGAGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSL SCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0240_protrusion
(SEQ ID NO: 205) DIQLTQSPSSLSASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNPGSGVPSRFSGSGSGTDFTISSLQPEDFATYYCQQSKEVPYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYL QMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0240_cavity
(SEQ ID NO: 206) DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTTYWMQWVRQAPGKGLEWIGAIYPGDGMTTYTQKFKDRFTISADKAKNS AYLQMNSLRAEDTAVYYCVRKGMDYWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0247_protrusion
(SEQ ID NO: 210) DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKPPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0247_cavity
(SEQ ID NO: 211) DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNSAY LQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1478_protrusion;
GUCY2C-1608_protrusion
(SEQ ID NO: 216) DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1478_cavity
(SEQ ID NO: 217) DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNVHEKFKNRFTISVDKAK NSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1608_cavity
(SEQ ID NO: 220) DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0250_protrusion
(SEQ ID NO: 248) DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-0250_cavity
(SEQ ID NO: 249) DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSRKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1678_protrusion
(SEQ ID NO: 282) DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1678_cavity
(SEQ ID NO: 283) DIQMTQSPSSLSASVGDRVTITCTSDQSLFNVRSGKNYLAWYQQKPGKAPKLLIYWASDRESGVPSRFSGSGSGTDFTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1679_protrusion
(SEQ ID NO: 284) DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1679_cavity
(SEQ ID NO: 285) DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSGKNYLAWYQQKPGKAPKLLIYWASDRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1680_protrusion
(SEQ ID NO: 286) DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG GUCY2C-1680_cavity
(SEQ ID NO: 287) DIQMTQSPSSLSASVGDRVTITCTSSESLFNVRSGKNYLAWYQQKPGKAPKLLIYWASDRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAY LQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

One way to determine the binding affinity of antibodies against GUCY2c or CD3 is to measure the binding affinity of monofunctional Fab fragments of the antibody. To produce monofunctional Fab fragments, the antibody (eg, IgG) can be digested with papain or expressed recombinantly. Anti-GUCY2c Fab affinity can be determined by surface plasmon resonance (using the BIACORE™ Surface Plasmon Resonance (SPR) System 3000™, BIACORE™, INC, Piscataway NJ) using pre-immobilized streptavidin (SA) sensor chips or using anti-mouse Fc antibody or anti-human Fc antibody and HBS-EP running buffer (0.01 M HEPES, pH 7, 4, 0.15 M NaCl, 3 mM EDTA, 0.005% v/v surfactant P20). Biotinylated or Fc-fused human GUCY2c can be diluted in HBS-EP buffer to a concentration of less than 0.5 μg/ml and injected into individual channels of the chip using variable contact times to obtain two ranges of antigen densities, 50-200 response units (RU) for detailed kinetics studies or 800-1000 RU for screening assays. Regeneration studies have shown that with 25 mM NaOH at 25% v/v. ethanol efficiently removes bound Fab while maintaining GUCY2c activity on the chip over 200 injections. Typically, serial dilutions (spanning concentrations ranging from 0.1 to 10 times the K _D ) of purified Fab samples are injected over 1 min at 100 μL/min and allow dissociation times of up to 2 h. Fab protein concentrations are determined by ELISA and/or SDS-PAGE using a known concentration of Fab (as determined by amino acid analysis) as a standard. Kinetic data on the rate of association (k _on ) and rate of dissociation (k _off ) are obtained simultaneously through a global fit to the 1:1 Langmuir binding model (Karlsson, R. Roos, H. Fagerstam, L. Petersson, B., Methods Enzymology 6. 99-110, 1994) using BIAevaluation software. The values of the equilibrium dissociation constant (K _D ) are calculated as k _off /k _on . This method is suitable for use in determining the binding affinity of an antibody to any GUCY2c, including human GUCY2c, other mammalian GUCY2c (such as mouse GUCY2c, rat GUCY2c or primate GUCY2c), as well as various forms of GUCY2c (for example, glycosylated GUCY2c). Antibody binding affinity is typically measured at 25°C, but can also be measured at 37°C.

Antibodies presented in the present description can be obtained by any method known in this field. To generate hybridoma cell lines, the route and schedule for immunizing host animals generally follows established and conventional methods for antibody stimulation and production, as further provided herein. General methods for producing human and mouse antibodies are known in the art and/or are presented herein.

It is contemplated that any mammal, including humans, or antibody-producing cells therefrom can be manipulated to form the basis for the production of mammalian cell lines, including human cell lines and hybridoma cell lines. Typically, the host animal is inoculated intraperitoneally, intramuscularly, orally, subcutaneously, intraplantarly, and/or intradermally with the amount of immunogen provided herein.

Hybridomas can be prepared from lymphocytes and immortalized myeloma cells using the general somatic cell hybridization method from Kohler, B. and Milstein, C., Nature 256:495-497, 1975 or a modified version thereof from Buck, D. W., et al., In Vitro , 18:377-381, 1982. Available myeloma lines can be used in the hybridization, including, but not limited to, X63-Ag8.653 and cells from the Salk Institute, Cell Distribution Center, San Diego, Calif., USA. Typically, the method involves fusing myeloma cells and lymphoid cells using a fusogen such as polyethylene glycol or by electrical methods well known to those skilled in the art. After fusion, the cells are separated from the fusion medium and grown in a selective growth medium, such as hypoxanthine-aminopterin-thymidine (HAT) medium, to eliminate unhybridized parent cells. Any of the media presented herein, supplemented or not supplemented with serum, can be used for the cultivation of hybridomas secreting monoclonal antibodies. As another alternative to the cell fusion method, EBV immortalized B cells can be used to produce the monoclonal antibodies of the present invention. Hybridomas are grown and subcloned, if desired, and supernatants are assayed for anti-immunogenic activity by conventional immunoassay techniques (eg, radioimmunoassay, enzyme immunoassay, or fluorescence immunoassay).

Hybridomas that can be used as a source of antibodies include all derivatives, progeny cells of parental hybridomas that produce monoclonal antibodies specific for GUCY2c, CD3, or parts thereof.

Hybridomas producing such antibodies can be grown in vitro or in vivo by known methods. Monoclonal antibodies can be isolated from culture media or body fluids by conventional immunoglobulin purification methods, if desired, such as ammonium sulfate precipitation, gel electrophoresis, dialysis, chromatography and ultrafiltration. Undesired activity, if present, can be eliminated, for example, by passing the drug through adsorbents derived from the immunogen attached to the solid phase and eluting or releasing the desired antibodies from the immunogen. A population of antibodies (e.g., monoclonal antibodies) can be generated by immunizing a host animal using human GUCY2c or CD3, or a fragment thereof, containing the target amino acid sequence conjugated to a protein that is immunogenic for the species to be immunized, e.g., limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent such as maleimidobenzoylsulfosuccinimide ester (conjugation via cysteine residues), N-hydroxysuccinimide (via lysine residues), glutaraldehyde, succinic anhydride, SOCl ₂ , or R ¹ N=C =NR, where R and R ¹ are different alkyl groups.

If desired, the antibody of interest (monoclonal or polyclonal) can be sequenced, and then the polynucleotide sequence can be cloned into an expression or propagation vector. The coding sequence for the antibody of interest can be maintained in a vector in a host cell, and the host cell can then be grown and frozen for later use. The production of recombinant monoclonal antibodies in cell culture can be accomplished by cloning antibody genes from B cells by methods known in the art. See, for example, Tiller et al., J. Immunol. Methods, 3291122008; US Patent No. 7314622.

Alternatively, the polynucleotide sequence can be used for genetic manipulation to humanize the antibody or to improve the affinity or other characteristics of the antibody. For example, the constant region can be designed to resemble human constant regions as closely as possible to avoid an immune response if the antibody is used in human clinical trials and treatment. It may be desirable to genetically manipulate the antibody sequence to achieve higher affinity for GUCY2c or CD3 and greater efficacy in inhibiting GUCY2c.

There are four general steps in the humanization of a monoclonal antibody. They are: (1) determination of the nucleotide and predicted amino acid sequence of the initial variable regions of the light and heavy chains of the antibody, (2) design of the humanized antibody, i.e. deciding which antibody framework to use during humanization, (3) specific humanization methods, and (4) transfection and expression of the humanized antibody. See, for example, US Patent Nos. 4816567, 5807715, 5866692, 6331415, 5530101, 5693761, 5693762, 5585089 and 6180370.

A number of humanized antibody molecules containing an antigen binding region derived from non-human immunoglobulin are described, including chimeric antibodies having rodent V regions or modified rodent V regions and corresponding CDRs fused to human constant regions. See, for example, Winter et al. Nature 349:293-299, 1991, Lobuglio et al. Proc. Nat. Acad. Sci. USA 86:4220-4224, 1989, Shaw et al. J Immunol. 138:4534-4538, 1987, and Brown et al. Cancer Res. 47:3577-3583, 1987. Elsewhere, rodent CDRs are described grafted onto a human framework region (FR) before fusion with a suitable human antibody constant region. See, for example, Riechmann et al. Nature 332:323-327, 1988, Verhoeyen et al. Science 239:1534-1536, 1988, and Jones et al. Nature 321:522-525, 1986. Another reference describes rodent CDRs supported by recombinantly engineered rodent framework regions. See, for example, European Patent Publication No. EP0519596. These “humanized” molecules are designed to minimize unwanted immune response to rodent anti-human antibody molecules, thereby limiting the duration and effectiveness of therapeutic use of these substances in human recipients. For example, the constant region of an antibody can be designed such that it is immunologically inert (eg, does not trigger complement-mediated lysis). See, for example, PCT patent application No. PCT/GB99/01441; UK Patent Application No. 9809951.8. Other methods for humanizing antibodies that can also be used are described in Daugherty et al., Nucl. Acids Res. 19:2471-2476, 1991, in US Patent Nos. 6180377, 6054297, 5997867, 5866692, 6210671 and 6350861 and PCT Publication No. WO01/27160.

The general principles regarding humanized antibodies described above can also be used to customize antibodies for use in, for example, dogs, cats, primates, horses and cattle. In addition, one or more aspects of humanization of an antibody provided herein can be combined, for example, CDR grafting, framework mutations, and CDR mutations.

In one embodiment, fully human antibodies can be produced using commercially available mice engineered to express specific human immunoglobulin proteins. Transgenic animals produced to achieve a more desirable (eg, fully human antibody) or more durable immune response can also be used to produce humanized or human antibodies. Examples of such technology include Xenomouse™ from Abgenix, Inc. (Fremont, CA) and HuMAb-Mouse® and TC Mouse™ from Medarex, Inc. (Princeton, NJ).

Alternatively, antibodies can be produced recombinantly and expressed by any method known in the art. As another alternative, antibodies can be produced recombinantly using phage display technology. See, for example, US Pat. Nos. 5,565,332, 5,580,717, 5,733,743 and 6,265,150 and Winter et al., Annu. Rev. Immunol. 12:433-455, 1994. Alternatively, phage display technology (McCafferty et al., Nature 348:552-553, 1990) can be used to produce human antibodies and antibody fragments in vitro from the immunoglobulin variable region (V) gene repertoire from immunized donors. In this method, antibody V domain genes are cloned in frame into the major or minor coat protein gene of a filamentous bacteriophage, such as M13 or fd, and displayed as functional antibody fragments on the surface of the phage particle. Because the filamentous phage particle contains a single-stranded copy of the DNA of the phage genome; selection based on the functional properties of the antibody also leads to selection of the gene encoding the antibody exhibiting these properties. In this way, the phage mimics some of the properties of a B cell. Phage display can be carried out in a variety of formats; for a review, see, for example, Johnson, Kevin S. and Chiswell, David J., Current Opinion in Structural Biology 3:564-571, 1993. Several sources of V gene segments can be used for phage display. Clackson et al., Nature 352:624-628, 1991 isolated a diverse set of anti-oxazolone antibodies from a small random combinatorial library of V genes obtained from the spleens of immunized mice. A repertoire of V genes from non-immunized human donors can be engineered and antibodies can be isolated against a diverse set of antigens (including self-antigens) essentially by the methods described in Mark et al., J. Mol. Biol. 222:581-597, 1991, or Griffith et al., EMBO J. 12:725-734, 1993. During the natural immune response, antibody genes accumulate mutations in large numbers (somatic supermutation). Some changes made will impart higher affinity, and B cells exhibiting high affinity surface immunoglobulin will preferentially divide and differentiate during subsequent antigen stimulation. This natural process can be imitated in a manner known as "chain shuffling" (Marks et al., Bio/Technol. 10:779-783, 1992). In this method, the affinity of human "primary" antibodies produced by phage display can be improved by sequentially replacing heavy and light chain V region genes with a repertoire of naturally occurring V domain gene variants obtained from non-immunized donors. This method makes it possible to obtain antibodies and antibody fragments with affinities in the pM-nM range. A strategy for obtaining a very large repertoire of phage antibodies (also known as “universal mother libraries”) is described in Waterhouse et al., Nucl. Acids Res. 21:2265-2266, 1993. Gene shuffling can also be used to produce human antibodies from rodent antibodies, where the human antibody has similar affinities and specificities to the parent rodent antibody. In this method, also referred to as “epitope imprinting,” a rodent antibody heavy or light chain V domain gene produced by phage display is replaced with a repertoire of human V domain genes, producing rodent-human chimeras. Selection using antigen results in the isolation of human variable regions capable of restoring a functional antigen-binding site, i.e. the epitope regulates (imprints) partner choice. When the method is repeated to replace the remaining rodent V domain, a human antibody is obtained (see PCT Publication No. WO93/06213). In contrast to the conventional humanization of rodent antibodies by CDR grafting, this method produces fully human antibodies without rodent scaffold or CDR residues.

Antibodies can be produced recombinantly by first isolating the antibodies and antibody-producing cells from host animals, obtaining the gene sequence, and using the gene sequence to recombinantly express the antibody in host cells (eg, CHO cells). Another method that can be used is to express the antibody sequence in plants (eg tobacco) or transgenic milk. Methods for recombinant expression of antibodies in plants or milk are described. See, for example, Peeters, et al. Vaccine 19:2756, 2001; Lonberg, N. and D. Huszar Int. Rev. Immunol 13:65, 1995; and Pollock, et al., J Immunol Methods 231:147, 1999. Methods for producing antibody derivatives, such as humanized, single chain, etc., are known in the art.

Immunoassays and flow cytometry-based sorting methods, such as fluorescence-activated cell sorting (FACS), can also be used to isolate antibodies specific for GUCY2c, CD3, or antigens of interest.

The antibodies provided herein can be coupled to a variety of different solid supports or carriers. Such supports may be active and/or inert. Well-known supports include polypropylene, polystyrene, polyethylene, dextran, nylon, amylases, glass, natural and modified celluloses, polyacrylamides, agaroses and magnetite. For purposes of the invention, the support may be soluble or insoluble. Those skilled in the art will know other suitable substrates for binding antibodies or will be able to determine them using routine experimentation. In some embodiments, the support contains a moiety that targets the myocardium.

DNA encoding monoclonal antibodies is readily isolated and sequenced by conventional methods (eg, using oligonucleotide probes that can specifically bind to genes encoding the heavy and light chains of monoclonal antibodies). Hybridoma cells serve as the preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors (such as the expression vectors described in PCT Publication No. WO87/04462), which are then transfected into host cells such as E. coli cells, monkey COS cells, CHO cells or myeloma cells. which do not otherwise produce immunoglobulin protein, to achieve the synthesis of monoclonal antibodies in recombinant host cells. See, for example, PCT Publication No. WO87/04462. The DNA can also be modified, for example, by replacing the coding sequence of the human heavy and light chain constant regions with homologous mouse sequences, Morrison et al., Proc. Nat. Acad. Sci. 81:6851, 1984, or by covalently linking to the immunoglobulin coding sequence all or part of the coding sequence of a non-immunoglobulin polypeptide. Thus, "chimeric" or "hybrid" antibodies are produced having the binding specificity of the monoclonal antibody of the present invention.

Antibodies against GUCY2c, CD3 or other antigens provided herein can be identified or characterized by methods known in the art, wherein the reduction in expression levels of GUCY2c, CD3 or other antigen is determined and/or measured. In some embodiments, an anti-GUCY2c antibody is identified by incubating the candidate agent with GUCY2c and monitoring the binding and/or concomitant reduction in GUCY2c expression levels. Binding assays can be performed using purified GUCY2c polypeptides or cells naturally expressing or transfected to express GUCY2c polypeptides. In one embodiment, the binding assay is a competitive binding assay that evaluates the ability of a candidate antibody to compete with a known anti-GUCY2c antibody for binding to GUCY2c. The assay can be performed in a variety of formats, including ELISA format.

Following initial identification, the activity of a candidate antibody against GUCY2c, CD3 or other antigen can be further confirmed and refined through biological assays known to test target biological activities. Alternatively, biological assays can be used to directly screen candidates. Some of the methods for identifying and characterizing antibodies are described in detail in the examples.

Antibodies against GUCY2c, CD3 or other antigen can be characterized by methods well known in this field. For example, one method is to identify the epitope to which it binds, or epitope mapping. There are many methods known in the art for mapping and characterizing the localization of epitopes on proteins, including crystal structure resolution of the antibody-antigen complex, competition assays, gene fragment expression assays, and synthetic peptide-based assays, as described, for example, in Chapter 11 of Harlow and Lane. Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1999. In an additional example, epitope mapping can be used to determine the sequence to which an antibody binds. Epitope mapping is commercially available from various sources, for example Pepscan Systems (Edelhertweg 15, 8219 PH Lelystad, The Netherlands). The epitope may be a linear epitope, i.e. contained in one fragment of amino acids, or a conformational epitope formed by the three-dimensional interaction of amino acids that may not be contained in one fragment. Peptides of various lengths (eg, at least 4-6 amino acids) can be isolated or synthesized (eg, recombinantly) and used for binding assays with an antibody against GUCY2c, CD3, or other antigen. In another example, the epitope to which an anti-GUCY2c, CD3, or other antigen antibody binds can be determined by a systemic screen using overlapping peptides derived from the GUCY2c, CD3, or other antigen sequence and determining binding to an anti-GUCY2c, CD3, or other antigen antibody. In gene fragment expression assays, an open reading frame encoding GUCY2c, CD3 or other antigen is fragmented randomly or using specific genetic constructs, and the reactivity of the expressed fragments of GUCY2c, CD3 or other antigen and the antibody to be tested is determined. Gene fragments can be obtained, for example, by PCR, and then transcribed and translated into protein in vitro in the presence of radioactive amino acids. The binding of the antibody to radiolabeled fragments of GUCY2c, CD3, or other antigen is then determined by immunoprecipitation and gel electrophoresis. Some epitopes can also be identified using large libraries of random peptide sequences displayed on the surface of phage particles (phage libraries). Alternatively, a defined library of overlapping peptide fragments can be tested for binding to a test antibody in simple binding assays. In a further example, antigen binding domain mutagenesis, domain shuffling experiments, and alanine scan mutagenesis can be performed to identify residues required, sufficient, and/or essential for epitope binding. For example, domain shuffling experiments can be performed using a mutant GUCY2c, CD3, or other antigen, where different fragments of GUCY2c, CD3, or another antigenic protein are replaced (rearranged) with sequences from GUCY2c of another species (e.g., mouse) or a closely related but antigenically distinct individual protein (for example, Trop-1). By assessing the binding of an antibody to a mutant GUCY2c, CD3, or other antigen, the importance of a particular fragment of GUCY2c, CD3, or other antigen for antibody binding can be assessed.

Another method that can be used to characterize an antibody against GUCY2c, CD3 or another antigen is the use of competition assays with other antibodies that are known to bind to the same antigen, i.e. different fragments on GUCY2c, CD3 or other antigen, to determine whether an antibody against GUCY2c, CD3 or other antigen binds to the same epitope as other antibodies, respectively. Competitive assays are well known to those skilled in the art.

An expression vector can be used to regulate the expression of an antibody against GUCY2c, CD3 or other antigen. One skilled in the art will be familiar with the introduction of expression vectors to achieve exogenous protein expression in vivo . See, for example, US Patent Nos. 6,436,908, 6,413,942 and 6,376,471. Administration of expression vectors includes local or systemic administration, including injection, oral administration, bioballistic gun or catheter administration, and topical administration. In another embodiment, the expression vector is introduced directly into the sympathetic trunk or ganglion or coronary artery, atrium, ventricle or pericardium.

Targeted delivery of therapeutic compositions containing an expression vector or subgenomic polynucleotides can also be used. Receptor-mediated methods of DNA delivery are described, for example, in Findeis et al., Trends Biotechnol., 11:202, 1993; Chiou et al., Gene Therapeutics: Methods And Applications Of Direct Gene Transfer, J.A. Wolff, ed., 1994; Wu et al., J. Biol. Chem., 263:621, 1988; Wu et al., J. Biol. Chem., 269:542, 1994; Zenke et al., Proc. Natl. Acad. Sci. USA, 87:3655, 1990; and Wu et al., J. Biol. Chem., 266:338, 1991. Therapeutic compositions containing polynucleotide are administered in the range of from about 100 ng to about 200 mg of DNA for local administration in a gene therapy method. The gene therapy method can also use concentration ranges from about 500 ng to about 50 mg, from about 1 μg to about 2 mg, from about 5 μg to about 500 μg, and from about 20 μg to about 100 μg of DNA. Therapeutic polynucleotides and polypeptides can be delivered using gene delivery vehicles. The gene delivery vehicle may be of viral or non-viral origin (see generally Jolly, Cancer Gene Therapy, 1:51, 1994; Kimura, Human Gene Therapy, 5:845, 1994; Connelly, Human Gene Therapy, 1:185 , 1995; and Kaplitt, Nature Genetics, 6:148, 1994). Expression of such coding sequences can be induced using endogenous mammalian promoters or heterologous promoters. Expression of a coding sequence may be constitutive or regulated.

Viral vectors for delivery of the desired polynucleotide and expression in the desired cell are well known in the art. Non-limiting examples of virus-based carriers include recombinant retroviruses (see, for example, PCT Publication Nos. WO90/07936; WO94/03622; WO93/25698; WO93/25234; WO93/11230; WO93/10218; WO91/02805; US Patent Nos. Nos. 5,219,740 and 4,777,127; UK Patent No. 2200651; and EP Patent No. EP0 345 242), alphavirus vectors (e.g., Sindbis virus vectors, Semliki Forest virus (ATCC VR-67; ATCC VR-1247), Ross River fever virus ( ATCC VR-373; ATCC VR-1246) and Venezuelan equine encephalitis virus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCC VR-532)) and adeno-associated virus (AAV) vectors (see e.g. , PCT Publications Nos. WO94/12649, WO93/03769; WO93/19191; WO94/28938; WO95/11984 and WO95/00655). Injection of DNA associated with a killed adenovirus can also be used, as described in Curiel, Hum. Gene Ther., 3:147, 1992.

Non-viral delivery vehicles and methods may also be used, including, but not limited to, polycation-fused DNA bound or unbound to killed adenovirus alone (see, e.g., Curiel, Hum. Gene Ther., 3:147, 1992) ; ligand-bound DNA (see, for example, Wu, J. Biol. Chem., 264:16985, 1989); eukaryotic cell carriers for delivery (see, for example, US Patent No. 5814482; PCT Publications Nos. WO95/07994; WO96/17072; WO95/30763 and WO97/42338) and nuclear charge neutralization or fusion with cell membranes. Deproteinized DNA can also be used. Examples of methods for incorporating deproteinized DNA are described in PCT Publication No. WO90/11092 and US Patent No. 5,580,859. Liposomes that can act as carriers for gene delivery are described in US Pat. No. 5,422,120; PCT publications No. WO95/13796; WO94/23697; WO91/14445; and European Publication No. EP 0524968. Additional approaches are described in Philip, Mol. Cell Biol., 14:2411, 1994 and Woffendin, Proc. Natl. Acad. Sci., 91:1581, 1994.

In some embodiments, the present invention relates to compositions, including pharmaceutical compositions, containing the antibodies of the invention described herein or prepared by the methods presented herein and having the characteristics described herein. Within the scope of the invention, pharmaceutical compositions may contain one or more antibodies that bind to GUCY2c, one or more bispecific antibodies that bind to CD3 and the tumor antigen GUCY2c, and/or one or more polynucleotides containing sequences encoding one or more of these antibodies. These compositions may further contain suitable excipients, such as pharmaceutically acceptable excipients, including buffers, well known in the art.

The invention also relates to methods for producing any of such antibodies. The antibodies of the present invention can be produced by methods known in the art. Polypeptides can be produced by proteolytic or other degradation of antibodies, by recombinant methods (ie, as single or fused polypeptides) as described above, or by chemical synthesis. Antibody polypeptides, particularly shorter polypeptides of up to about 50 amino acids, are conveniently produced by chemical synthesis. Chemical synthesis methods are known in the art and are commercially available. For example, the antibody can be produced using an automated polypeptide synthesizer using a solid phase method. Also see, US Patent Nos. 5807715, 4816567 and 6331415.

Heteroconjugated antibodies, comprising two covalently linked antibodies, are also within the scope of the invention. Such antibodies are used to target immune system cells to unwanted cells (US Patent No. 4676980) and to treat HIV infection (PCT Publication Nos. WO91/00360 and WO92/200373; and European Patent Publication No. EP03089). Heteroconjugated antibodies can be produced by any convenient cross-linking methods. Suitable crosslinking agents and methods are well known in the art and are described in US Pat. No. 4,676,980.

Chimeric or hybrid antibodies can also be produced in vitro by known methods of synthetic protein chemistry, including methods involving cross-linking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or through thioester bond formation. Examples of suitable reagents for this purpose include iminothiolate and methyl 4-mercaptobutyrimidate.

In recombinant humanized antibodies, the Fcγ portion can be modified to avoid interaction with the Fcγ receptor and complement and the immune system. Methods for producing such antibodies are described in PCT Publication No. WO99/58572. For example, the constant region can be designed to more closely resemble human constant regions to avoid an immune response if the antibody is used in human clinical trials and treatment. See, for example, US Patent Nos. 5,997,867 and 5,866,692.

The anti-GUCY2c antibodies and anti-CD3-GUCY2c bispecific antibodies provided herein may contain one or more amino acid substitutions, insertions and/or deletions in the framework regions and/or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences , from which antibodies are obtained. Such mutations can be established by comparing the amino acid sequences presented herein with germline sequences available, for example, in publicly available antibody sequence databases. The present invention relates to antibodies and antigen binding fragments thereof derived from any of the amino acid sequences presented herein, wherein one or more amino acids in one or more framework regions and/or CDR regions have been mutated to the corresponding residues of the germline sequence from which the antibody was derived , or to corresponding residues of another human germline sequence, or to a conservative amino acid substitution of corresponding germline residues (such sequence changes are collectively referred to herein as “germline mutations”). A person skilled in the art, starting with the heavy and light chain variable region sequences presented herein, can easily produce numerous antibodies and antigen binding fragments containing one or more individual germline mutations or combinations thereof. In some embodiments, all of the framework residues and/or CDRs in the VH and/or VL domains are back-mutated to residues found in the original germline sequence from which the antibody was derived. In other embodiments, only certain residues are backmutated to the original germline sequence, for example, only the mutated residues found in the first 8 amino acids of FR1 or the last 8 amino acids of FR4, or only the mutated residues found in CDR1, CDR2, or CDR3. In other embodiments, one or more residues from the framework and/or CDR are mutated into corresponding residues from a different germline sequence (ie, a germline sequence different from the germline sequence from which the antibody was originally derived).

In addition, the antibodies of the present invention may contain any combination of two or more germline mutations in framework regions and/or CDR regions, for example, where some individual residues are mutated to the corresponding residue of a particular germline sequence, while some other residues different from the original germline sequence are retained or mutated to the corresponding residue of another germline sequence. Once produced, antibodies and antigen-binding fragments containing one or more germline mutations can be readily tested for one or more desired properties, such as improved binding specificity, increased binding affinity, improved or increased antagonistic or agonistic biological properties (respectively), reduced immunogenicity, and etc. Antibodies and antigen binding fragments produced by this general method are included in the present invention.

The present invention also provides anti-GUCY2c antibodies and anti-CD3-GUCY2c bispecific antibodies containing variants of any of the HC VR, LC VR and/or CDR amino acid sequences described herein having one or more conservative substitutions. For example, the present invention relates to anti-GUCY2c antibodies and anti-CD3-GUCY2c bispecific antibodies having amino acid sequences HC VR, LC VR and/or CDRs with, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc. .d. conservative amino acid substitutions relative to any of the amino acid sequences HC VR, LC VR and/or CDR, as presented in the present description.

Thus, the present invention relates to modifications of variants of the antibodies and polypeptides of the invention as described herein, including functionally equivalent antibodies that do not significantly affect their properties, and variants having increased or decreased activity and/or affinity. For example, the amino acid sequence can be mutated to produce an antibody with the desired binding affinity for GUCY2c and/or CD3. Modification of polypeptides is a common practice in this field and does not require detailed discussion herein. Examples of modified polypeptides include polypeptides with conservative substitutions of amino acid residues, one or more deletions or additions of amino acids that do not significantly, adversely change functional activity or that mature (increase) the affinity of the polypeptide for its ligand, or the use of chemical analogues.

Amino acid sequence insertions include amino- and/or carboxy-terminal fusions ranging in length from a single residue to polypeptides containing a hundred or more residues, as well as insertions of single or multiple amino acid residues into a sequence. Examples of terminal insertions include an antibody with an N-terminal methionyl residue or an antibody fused to an epitope tag. Other insertional variants of an antibody molecule include fusion of an enzyme or polypeptide that increases the half-life of the antibody in the bloodstream to the N- or C-terminus of the antibody.

Substitution options include the removal of at least one amino acid residue in the antibody molecule and the insertion of another residue in its place. The regions of greatest interest for replacement mutagenesis include the hypervariable regions, but FR changes are also contemplated. Conservative substitutions are listed in Table 8 under the heading “Conservative Substitutions.” If such substitutions result in a change in biological activity, then more significant changes, designated in Table 8 as “example substitutions” or described below in relation to amino acid classes, can be introduced and the resulting products screened.

Table 8

Original residue (naturally occurring amino acid) Conservative Substitutions Examples of substitutions Ala (A) Val Val; Leu; Ile Arg(R) Lys Lys; Gln; Asn Asn(N) Gln Gln; His; Asp, Lys; Arg Asp (D) Glu Glu; Asn Cys(C) Ser Ser; Ala Gln(Q) Asn Asn; Glu Glu (E) Asp Asp; Gln Gly (G) Ala Ala His(H) Arg Asn; Gln; Lys; Arg Ile (I) Leu Leu; Val; Met; Ala; Ph; norleucine Leu (L) Ile Norleucine; Ile; Val; Met; Ala; Phe Lys (K) Arg Arg; Gln; Asn Met(M) Leu Leu; Ph; Ile Phe (F) Tyr Leu; Val; Ile; Ala; Tyr Pro (P) Ala Ala Ser (S) Thr Thr Thr (T) Ser Ser Trp(W) Tyr Tyr; Phe Tyr (Y) Phe Trp; Ph; Thr; Ser Val(V) Leu Ile; Leu; Met; Ph; Ala; norleucine

Significant modifications of the biological properties of an antibody are accomplished by selecting substitutions that differ significantly in their effects with respect to (a) the backbone structure of the polypeptide at the site of the substitution, such as sheet or helix conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) side chain volume. Natural amino acid residues are divided into groups taking into account the general properties of the side chain:

(1) Non-polar: norleucine, Met, Ala, Val, Leu, Ile;

(2) Polar uncharged: Cys, Ser, Thr, Asn, Gln;

(3) Acidic (negatively charged): Asp, Glu;

(4) Basic (positively charged): Lys, Arg;

(5) Residues affecting chain orientation: Gly, Pro; And

(6) Aromatic: Trp, Tyr, Phe, His.

Non-conservative substitutions are made by replacing a member of one of these classes with a member of another class.

Any cysteine residue not involved in maintaining proper antibody conformation can also be replaced, typically with serine, to improve the oxidative stability of the molecule and prevent abnormal cross-linking. Conversely, cysteine linkages can be added to an antibody to improve its stability, particularly if the antibody is an antibody fragment such as an Fv fragment.

Amino acid modifications can range from changing or modifying one or more amino acids to complete the remodeling of a region, such as a variable region. Changes in the variable region may result in changes in binding affinity and/or specificity. In some embodiments, no more than one to five conservative amino acid substitutions are made within the CDR domain. In other embodiments, no more than one to three conservative amino acid substitutions are made within the CDR domain. In other embodiments, the CDR domain is CDR3 VH and/or CDR3 VL.

Modifications also include glycosylated and non-glycosylated polypeptides, as well as polypeptides with other post-translational modifications, such as glycosylation with various sugars, acetylation and phosphorylation. Antibodies are glycosylated at conserved positions in their constant regions (Jefferis and Lund, Chem. Immunol. 65:111-128, 1997; Wright and Morrison, TibTECH 15:26-32, 1997). The oligosaccharide side chains of immunoglobulins influence protein function (Boyd et al., Mol. Immunol. 32:1311-1318, 1996; Wittwe and Howard, Biochem. 29:4175-4180, 1990) and intramolecular interactions between parts of the glycoprotein that can influence on conformation and represent the three-dimensional surface of the glycoprotein (Jefferis and Lund, supra; Wyss and Wagner, Current Opin. Biotech. 7:409-416, 1996). Oligosaccharides can also serve to target the specified glycoprotein to certain molecules, taking into account specific recognition structures. Antibody glycosylation has also been reported to influence antibody-dependent cellular cytotoxicity (ADCC). In particular, CHO cells with tetracycline-regulated expression of β(1,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase that catalyzes the formation of GlcNAc at branch points, are reported to have improved ADCC activity (Umana et al., Mature Biotech. 17:176-180, 1999).

Antibody glycosylation is typically N-linked or O-linked. The term "N-linked" refers to the attachment of a carbohydrate molecule to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine, asparagine-X-threonine, and asparagine-X-cysteine, where X is any amino acid except proline, are recognition sequences for the enzymatic attachment of a carbohydrate molecule to the asparagine side chain. Thus, the presence of these tripeptide sequences in a polypeptide results in the formation of a potential glycosylation site. The term "O-linked" glycosylation refers to the addition of one of the sugars N-acetylgalactosamine, galactose or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine can also be used.

The addition of glycosylation sites to an antibody is conveniently accomplished by altering the amino acid sequence so that it contains one or more of the tripeptide sequences described above (in the case of N-linked glycosylation sites). The change can also be made by adding or replacing one or more serine or threonine residues in the sequence of the parent antibody (in the case of O-linked glycosylation sites).

The glycosylation profile of antibodies can also be changed without changing the underlying nucleotide sequence. Glycosylation mainly depends on the host cell used to express the antibody. Because Since the cell type used to express recombinant glycoproteins, such as antibodies, as potential therapeutic agents is rarely a native cell, variations in the glycosylation profile of antibodies can be expected (see, for example, Hse et al., J. Biol. Chem. 272: 9062-9070, 1997).

In addition to the choice of host cells, factors affecting glycosylation during recombinant antibody production include culture regime, media formulation, culture density, oxygenation, pH, purification schemes, etc. Various methods have been proposed to alter the glycosylation profile achieved in a particular host, including the incorporation or overexpression of certain enzymes involved in the production of oligosaccharides (US patent Nos. 5047335, 5510261 and 5278299). Glycosylation or some types of glycosylation can be enzymatically removed from a glycoprotein, for example, using endoglycosidase H (Endo H), N-glycosidase F, endoglycosidase F1, endoglycosidase F2, endoglycosidase F3. In addition, a recombinant host cell can be genetically engineered to be defective in processing certain types of polysaccharides. These and similar methods are well known in the art.

Other methods of modification include the use of coupling methods known in the art, including, but not limited to, enzymatic methods, oxidative replacement and chelation. The modifications can be used, for example, to attach labels for immunoassays. Modified polypeptides are prepared by methods conventional in the art and can be screened using standard methods known in the art, some of which are described below and in the Examples.

In some embodiments, the antibody comprises a modified constant region, such as a constant region having increased affinity for human Fc receptor gamma, that is immunologically inert or partially inert, e.g., does not trigger complement-mediated lysis, does not stimulate antibody-dependent cellular cytotoxicity (ADCC) or does not activate macrophages; or has reduced activities (compared to an unmodified antibody) in any one or more of the following: triggering complement-mediated lysis, stimulating antibody-dependent cellular cytotoxicity (ADCC), or activating microglia. To achieve the optimal level and/or combination of effector functions, various modifications of the constant region can be used. See, for example, Morgan et al., Immunology 86:319-324, 1995; Lund et al., J. Immunology 157:4963-9 157:4963-4969, 1996; Idusogie et al., J. Immunology 164:4178-4184, 2000; Tao et al., J. Immunology 143: 2595-2601, 1989; and Jefferis et al., Immunological Reviews 163:59-76, 1998. In some embodiments, the constant region is modified as described in Eur. J. Immunol., 29:2613-2624, 1999; PCT Application No. PCT/GB99/01441; and/or UK Application No. 9809951.8. In other embodiments, the constant region is aglycosylated at N-linked glycosylation. In some embodiments, the constant region is N-linked glycosylated as a result of mutation of the glycosylated amino acid residue or flanking residues that are part of the N-linked glycosylation recognition sequence in the constant region. For example, the N-glycosylation site of N297 can be mutated to A, Q, K or H. See, Tao et al., J. Immunology 143: 2595-2601, 1989; and Jefferis et al., Immunological Reviews 163:59-76, 1998. In some embodiments, the constant region is aglycosylated at N-linked glycosylation. The constant region may be aglycosylated at N-linked glycosylation enzymatically (eg, through carbohydrate removal by the enzyme PNase) or through expression in a glycosylation-deficient host cell.

Other modifications of antibodies include antibodies modified as described in PCT Publication No. WO99/58572. These antibodies contain, in addition to a binding domain directed against a target molecule, an effector domain having an amino acid sequence substantially homologous to all or part of the human immunoglobulin heavy chain constant region. These antibodies can bind to the target molecule without triggering significant complement-dependent lysis or cell-mediated resolution of the target. In some embodiments, the effector domain may specifically bind to FcRn and/or FcγRIIb. They are typically based on chimeric domains derived from two or more human immunoglobulin heavy chain CH2 domains. Antibodies modified in this manner are particularly suitable for use in chronic antibody therapy to avoid inflammatory and other adverse reactions to conventional antibody therapy.

The present invention includes affinity mature embodiments. For example, affinity mature antibodies can be produced by methods known in the art (Marks et al., Bio/Technology, 10:779-783, 1992; Barbas et al., Proc Nat. Acad. Sci, USA 91:3809-3813, 1994 Schier et al., Gene, 169:147-155, 1995; Yelton et al., J. Immunol., 155:1994-2004, 1995; Jackson et al., J. Immunol., 154(7):3310 -9, 1995, Hawkins et al., J. Mol. Biol., 226:889-896, 1992; and PCT Publication No. WO04/058184).

The following methods can be used to adjust antibody affinity and to characterize CDRs. One method of characterizing the CDR of an antibody and/or changing (such as improving) the binding affinity of a polypeptide, such as an antibody, is called "library scanning mutagenesis". Typically, scanning mutagenesis of a library is carried out as follows. One or more amino acid positions in the CDR are replaced by two or more (for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) amino acids using methods known in the art. In this way, small libraries of clones are obtained (in some embodiments, one for each amino acid position analyzed), each of which has a complexity of two or more members (if two or more amino acids are substituted at each position). Typically, the library also includes a clone containing the native (unsubstituted) amino acid. A small number of clones, for example approximately 20-80 clones (depending on the complexity of the library), from each library are screened for binding affinity to the target polypeptide (or other binding target) and candidates with increased, the same, decreased or no binding are identified. Methods for determining binding affinity are well known in the art. Binding affinity can be determined using the BIACORE™ surface plasmon resonance assay, which detects differences in binding affinity of approximately 2-fold or greater. BIACORE™ is particularly suitable if the parent antibody already binds with relatively high affinity, for example a KD of approximately 10 nM or less. Screening using BIACORE™ Surface Plasmon Resonance Assay described in the examples herein.

Binding affinity can be determined using the Kinexa biosensor, proximity scintillation assay, ELISA, ORIGEN immunoassay (IGEN), fluorescence quenching, fluorescence transfer, and/or yeast display. Binding affinity can also be screened using a suitable biological assay.

In some embodiments, each amino acid position in the CDR is replaced (in some embodiments, one at a time) with all 20 naturally occurring amino acids using mutagenesis techniques known in the art (some of which are presented herein). This results in small clone libraries (in some embodiments, one library per amino acid position analyzed), each having a complexity of 20 members (if all 20 amino acids are replaced at each position).

In some embodiments, the library to be screened contains substitutions at two or more positions, which may be in the same CDR or two or more CDRs. Thus, a library may contain substitutions at two or more positions in a single CDR. The library may contain a substitution at two or more positions in two or more CDRs. The library may contain a replacement at 3, 4, 5 or more positions, where the specified positions are found in two, three, four, five or six CDRs. Substitutions can be made using codons with low redundancy. See, for example, Table 2 from Balint et al., Gene 137(1):109-18, 1993.

Candidates with improved binding can be sequenced, thereby identifying a mutant with a substitution in the CDR resulting in improved affinity (also referred to as an "improved" substitution). Candidates that bind can also be sequenced, thereby identifying CDR replacements that preserve binding.

Multiple rounds of screening can be performed. For example, candidates (each containing an amino acid substitution at one or more positions of one or more CDRs) with improved binding are also suitable for the design of a second library containing at least the original and substituted amino acid at each improved CDR position (i.e., amino acid position in the CDR in which the substitution mutant exhibits improved binding). The preparation and screening or selection of this library is further described below.

Library scanning mutagenesis is also a way to characterize CDRs, since the frequency of clones with improved binding, the same binding, reduced binding, or no binding also provides information regarding the importance of each amino acid position for the stability of the antibody-antigen complex. For example, if a CDR position retains binding when changed across all 20 amino acids, that position is identified as a position that is unlikely to be required for antigen binding. Conversely, if a position in a CDR retains binding with only a small fraction of substitutions, that position is identified as a position important for CDR function. Thus, by scanning mutagenesis techniques, libraries obtain information regarding positions in the CDRs that can be changed into many different amino acids (including all 20 amino acids), and positions in the CDRs that cannot be changed or that can be changed into multiple amino acids.

Affinity-improved candidates may be combined in a second library comprising the improved amino acid, the parent amino acid at that position, and which may further include additional substitutions at that position, depending on the complexity of the library desired or allowed using the desired screening or selection method. In addition, if desired, a contiguous amino acid position can be randomized into at least two or more amino acids. Randomization of contiguous amino acids may allow additional conformational flexibility in the mutant CDR, which in turn may allow or facilitate the insertion of more enhancement mutations. The library may also contain substitutions at positions that do not show improved affinity in the first round of screening.

The second library is screened or selected for library members with improved and/or altered binding affinity using any method known in the art, including screening using the BIACORE™ surface plasmon resonance assay, and selection using any selection method known in the art, including phage display, yeast display and ribosomal display.

The present invention also provides compositions containing antibodies conjugated (eg, bound) to an agent that facilitates binding to a solid support (such as biotin or avidin). For simplicity, one can generally refer to antibodies with the understanding that these methods are used in relation to any of the embodiments of the anti-GUCY2c antibodies presented herein. The term "conjugation" generally refers to the binding of these components, as presented herein. Binding (typically representing the fixation of these components close to each other at least for administration) can be achieved in a number of ways. For example, a direct reaction between an agent and an antibody is possible if each component includes a substituent that is capable of reacting with the other substituent. For example, a nucleophilic group, such as an amino group or a sulfhydryl group, on one component may react with a carbonyl-containing group, such as an anhydride or acid halide, or an alkyl group containing a good leaving group (eg, a halide) on the other.

In some embodiments, the first and second polypeptides (and any additional polypeptides forming the bispecific antibody) are selected as a first step. Typically, the nucleic acid encoding these polypeptides must be isolated in such a way that it can be modified to encode a protrusion or a recess, or both, as defined herein. However, mutations can be introduced synthetically, for example, using a peptide synthesizer. Moreover, in case the substituted residue is a non-natural residue, the method of Noren et al., supra, is available for producing polypeptides having such substitutions. In addition, part of the bispecific antibody is suitably produced recombinantly in cell culture, and other parts of the molecule are obtained by the above methods.

Methods for isolating antibodies and producing bispecific antibodies are as follows. However, it should be understood that bispecific antibodies can be obtained from other polypeptides, or they can include other polypeptides, using methods known in this field. For example, a nucleic acid encoding a polypeptide of interest (eg, a ligand, receptor, or enzyme) can be isolated from a cDNA library obtained from tissue believed to contain and express the polypeptide's mRNA at a detectable level. Libraries are screened using probes (such as antibodies or oligonucleotides of approximately 20-80 bases) designed to identify the gene of interest or the protein it encodes. Screening of a cDNA or genomic library using a selected probe can be accomplished by standard methods as described in chapters 10-12 of Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989).

As stated above, the term "bispecific antibody" refers to a complex of two or more polypeptide chains, each containing at least one antibody VL region and one antibody VH region or fragment thereof, wherein the VL and VH regions in each polypeptide chain are derived from different antibodies. In particular aspects, the bispecific antibody includes dimers or tetramers of polypeptide chains containing VL and VH regions. Individual polypeptide chains containing multimeric proteins can be covalently linked to at least one other peptide of the multimer via interchain disulfide bonds.

The bispecific antibodies of the present invention simultaneously target T cells (CD3) and tumor cells (GUCY2c) and successfully direct and activate T cell cytotoxicity against tumor cells expressing GUCY2c.

Each polypeptide chain of a bispecific antibody contains a VL region and a VH region that can be covalently linked by a linker, where the linker is a glycine-serine linker containing glycine and serine residues such that the binding domains of the antibody are limited in self-assembly. In a specific embodiment, the glycine-serine linker is linker 1 comprising the sequence SEQ ID NO: 190. In addition, each polypeptide chain contains a heterodimerization domain that promotes heterodimerization and/or stabilization of multiple polypeptide chains and reduces the likelihood of homodimerization of different polypeptide chains. The heterodimerization domain may be located at the N-terminus of the polypeptide chain or the C-terminus. The heterodimerization domain may contain a cysteine linker (linker 2) having a length of 1, 2, 3, 4, 5, 6 or more amino acid residues. The interaction of two polypeptide chains can lead to the formation of two VL/VH pairs, forming two epitope-binding domains, i.e. bivalent molecule. Neither VH or VL region is restricted to any position in the polypeptide chain, i.e. is not limited to the amino terminus or the carboxy terminus, and the regions are also not limited in their positions relative to each other, i.e. the VL region may be N-terminal to the VH region and vice versa. The only limitation is that the complementary polypeptide chain be available to form a functional bispecific antibody. If the VL and VH regions are derived from antibodies specific for different antigens, the formation of a functional bispecific antibody requires the interaction of two different polypeptide chains, i.e. heterodimer formation. In contrast, if two different polypeptide chains are free to interact, for example in a recombinant expression system, one containing VLA and VHB (A being the first epitope and B being the second epitope) and the other containing VLB and VHA, the two different binding sites can form: VLA-VHA and VLB-VHB. In all pairs of polypeptide chains of a bispecific antibody, displacement or incorrect binding of the two chains is possible, for example, interaction of the VL-VL or VH-VH regions. However, the purification of functional bispecific antibodies is easy to achieve based on the immunospecificity of the properly dimerized binding site using any affinity-based method known in the art or described herein, such as affinity chromatography.

In one embodiment, the polypeptide chains of a bispecific antibody may contain various linkers and peptides. Linkers and peptides may contain 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or more amino acids. In some embodiments, the anti-GUCY2c VL and the anti-CD3 VH are connected using linker 1 or linker 2; and the anti-CD3 VL and the anti-GUCY2c VH are connected via linker 1 or linker 2. In some such embodiments, linker 1 comprises the sequence SEQ ID NO: 190. In other embodiments, linker 2 comprises the sequence SEQ ID NO: 191.

In some embodiments, domain 1 is covalently linked to a first heterodimerization-promoting domain via a cysteine linker, and domain 2 is covalently linked to a second heterodimerization-promoting domain via a cysteine linker. Each of the cysteine linkers includes at least one cysteine residue allowing the formation of an intramolecular disulfide bond. In further embodiments of the above, the cysteine linker contains at least five amino acids. In some such embodiments, the cysteine linker is linker 3 comprising the sequence SEQ ID NO: 192.

In some embodiments, the first polypeptide chain is covalently linked to the second polypeptide chain through at least one disulfide bond. In some such embodiments, at least one disulfide bond is formed between linker 3 of the first polypeptide chain and linker 3 of the second polypeptide chain. In another such embodiment, at least one disulfide bond is formed between the first heterodimerization promoting domain and the second heterodimerization promoting domain. In specific embodiments, each disulfide bond is formed by linking two cysteine residues.

The bispecific antibodies of the invention can simultaneously bind to two separate and distinct epitopes. In some embodiments, the at least one epitope binding site is specific for a CD3 determinant expressed on an immune effector cell, for example, expressed on T lymphocytes. In one embodiment, the bispecific antibody molecule binds to a determinant of the effector cell and also activates the effector cell.

In specific embodiments, the bispecific antibody of the present invention (a) binds to the extracellular domain of human GUCY2c, (b) exhibits an increased serum and tumor half-life of 30 minutes to 100 days; and/or (c) exhibits a lower EC ₅₀ value from 0.0001 nM to 100 nM in the presence of increased levels of GUCY2c expression or increased levels of receptor density.

In one embodiment, the epitope-binding domain can bind to a tumor-specific antigen GUCY2c associated with breast, ovarian, thyroid, prostate, cervical, lung cancer (including, but not limited to, non-small cell lung cancer and small cell lung cancer) , bladder, endometrium, head and neck, testicular and glioblastoma. In some embodiments, the cancer is a digestive system cancer selected from the group consisting of cancer of the esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile duct, and pancreas. In specific embodiments, the therapy activates a cytolytic T cell response.

The bispecific antibodies of the present invention contain antigen binding domains, typically derived from immunoglobulins or antibodies. The antibodies from which the binding domains used in the methods of the invention are derived can be obtained from any animal, including birds and mammals (e.g., human, non-human primate, mouse, donkey, sheep, rabbit, goat, guinea pig, camel, horse or chicken). Preferably, the antibodies are human or humanized monoclonal antibodies.

The bispecific antibodies of the present invention can be characterized in various ways. In particular, the molecules of the invention can be assayed for the ability to immunospecifically bind to an antigen. Molecules identified as immunospecifically binding to an antigen can then be analyzed for their specificity and affinity for the antigen.

The bispecific antibody of the present invention can be produced by various methods well known in the art, including de novo protein synthesis and recombinant expression of nucleic acids encoding binding proteins. The desired nucleic acid sequences can be obtained by recombinant methods (for example, by PCR mutagenesis of a previously obtained variant of the desired polynucleotide) or by solid-phase DNA synthesis. Typically, recombinant expression methods are used. In one aspect, the present invention provides a polynucleotide comprising a sequence encoding anti-CD3 VH and/or VL; in another aspect, the invention relates to a polynucleotide containing a sequence encoding VH and/or VL against GUCY2c. Due to the degeneracy of the genetic code, different nucleic acid sequences encode each immunoglobulin amino acid sequence, and the present invention relates to all nucleic acids encoding binding proteins presented herein.

The bispecific antibody of the present invention may contain two scFV domains binding to a target antigen and CD3 fused to an IgG Fc domain. In one embodiment, the bispecific antibody contains two scFV domains that bind to GUCY2c and CD3 epsilon fused to the Fc domain of IgG1. Specifically, Figures 1A, 1B, 1C and 1D show four alternative anti-CD3-GUCY2c bispecific antibody designs. The bispecific antibody contains a first heterodimerization promoting domain and a second heterodimerization promoting domain. Each first heterodimerization-promoting and second heterodimerization-promoting domain may comprise a CH2 domain and/or a CH3 domain of an immunoglobulin Fc region, wherein the CH2 domain and/or CH3 domain is modified to comprise a protrusion or a recess. Modifications to the Fc portion of the bispecific antibody are described below.

In an aspect of the present invention, a bispecific antibody, as shown in Figures 1A, 1B, 1C and 1D, contains a first polypeptide chain and a second polypeptide chain.

In one aspect, the present invention provides a bispecific antibody as described herein, wherein: (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-GUCY2c VL (anti-GUCY2c VL). GUCY2c) and VH anti-CD3 antibodies (VH anti-CD3), and (ii) the first heterodimerization-promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-CD3 VL (anti-CD3 VL) and anti-GUCY2c VH (anti-GUCY2c VH), and (ii) a second heterodimerization-promoting domain.

In another aspect, the invention provides a bispecific antibody as described herein, wherein: (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 comprising anti-CD3 VL and anti-GUCY2c VH, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-GUCY2c VL and anti-CD3 VH, and (ii) a second heterodimerization promoting domain.

In yet another aspect, the invention provides a bispecific antibody as described herein, wherein: (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-GUCY2c VH and anti-CD3 VL, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-CD3 VH and anti-GUCY2c VL, and (ii) a second heterodimerization promoting domain.

In another aspect, the invention provides a bispecific antibody as described herein, wherein: (a) the first polypeptide chain comprises, from N-terminus to C-terminus: (i) domain 1 containing anti-CD3 VH and anti-GUCY2c VL, and (ii) a first heterodimerization promoting domain; and (b) the second polypeptide chain contains, in an N-terminal to C-terminal direction: (i) domain 2 containing anti-GUCY2c VH and anti-CD3 VL and (ii) a second heterodimerization promoting domain.

In some embodiments of the above, the anti-GUCY2c VL and the anti-GUCY2c VH form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3.

In additional embodiments of any of the above, the bispecific antibody of the present invention comprises (a) an anti-GUCY2c VH CDR1 comprising the sequence SEQ ID NO: 12, 20, 27, 34, 42, 74, 257, 258, 259, 260, or 261 ; (b) CDR2 VH against GUCY2c containing the sequence SEQ ID NO: 13, 21, 28, 35, 43, 53, 66, 68, 70, 72, 75, 262, 263, 264, 265, 266 or 267; (c) CDR3 VH against GUCY2c containing the sequence SEQ ID NO: 14, 22, 29, 36 or 44; (d) CDR1 VL against GUCY2c containing the sequence SEQ ID NO: 93, 101, 105, 107, 113, 120, 148, 153, 163 or 167; (e) CDR2 VL against GUCY2c containing the sequence SEQ ID NO: 78, 94, 102, 108, 114, 141, 144, 146, 149, 151, 157, 159, 161, 164 or 168; and (f) CDR3 VL against GUCY2c containing the sequence SEQ ID NO: 95, 109, 115, 121, 142, 154, 165 or 169.

In a specific embodiment, the bispecific antibody of the present invention comprises (a) an anti-GUCY2c VH CDR1 comprising the sequence SEQ ID NO: 74; (b) CDR2 VH against GUCY2c containing the sequence SEQ ID NO: 75; (c) CDR3 VH against GUCY2c containing the sequence SEQ ID NO: 29; (d) CDR1 VL against GUCY2c containing the sequence SEQ ID NO: 148; (e) CDR2 VL against GUCY2c containing the sequence SEQ ID NO: 149; and (f) CDR3 VL against GUCY2c containing the sequence SEQ ID NO: 142.

In specific embodiments, the bispecific antibody comprises: (a) an anti-GUCY2c VH region comprising the sequence SEQ ID NO: 73; and (b) an anti-GUCY2c VL region containing the sequence shown in SEQ ID NO: 147.

In further embodiments, the present invention provides bispecific antibodies comprising (a) an anti-CD3 VH CDR1 comprising the sequence SEQ ID NO: 2; (b) CDR2 VH anti-CD3 containing the sequence SEQ ID NO: 3 or 10; (c) CDR3 VH anti-CD3 containing the sequence SEQ ID NO: 4; (d) VL anti-CD3 CDR1 containing the sequence SEQ ID NO: 77, 85, or 91; (e) CDR2 VL anti-CD3 containing SEQ ID NO: 78; and (f) an anti-CD3 VL CDR3 comprising the sequence SEQ ID NO: 79.

In a specific embodiment, the present invention provides a bispecific antibody comprising (a) an anti-CD3 VH CDR1 comprising the sequence SEQ ID NO: 2; (b) CDR2 VH anti-CD3 containing the sequence SEQ ID NO: 10; (c) CDR3 VH anti-CD3 containing the sequence SEQ ID NO: 4; (d) VL anti-CD3 CDR1 containing the sequence SEQ ID NO: 91; (e) VL anti-CD3 CDR2 containing the sequence SEQ ID NO: 78; and (f) an anti-CD3 VL CDR3 comprising the sequence SEQ ID NO: 79.

In one embodiment, the present invention provides a bispecific antibody comprising: (a) an anti-CD3 VH region comprising the sequence SEQ ID NO: 1, 9 or 273; and (b) an anti-CD3 VL region comprising the sequence SEQ ID NO: 76, 84, 90, 274, 275 or 276. In a specific embodiment, the bispecific antibody of the present invention comprises: (a) an anti-CD3 VH region comprising the sequence SEQ ID NO: 9; and (b) an anti-CD3 VL region comprising the sequence SEQ ID NO: 90.

In another aspect of the invention, linker 3 may comprise a truncated human IgG1 lower hinge region having the sequence GCPPCP (SEQ ID NO: 192) with at least one glycine residue preceding the lower hinge region.

In one aspect, the first heterodimerization promoting domain may comprise an Fc chain comprising a CH2 and/or CH3 domain modified to contain an overhang Fc chain comprising the sequence SEQ ID NO: 188, 196, 199, 202, 205, 210 , 216, or 248, 282, 284 or 286; or a cavity Fc chain comprising the sequence SEQ ID NO: 189, 197, 200, 203, 206, 211, 217, 220, 249, 283, 285, or 287. In another aspect, the second heterodimerization promoting domain may comprise an Fc chain, containing a CH2 and/or CH3 domain modified to contain an overhang Fc chain containing the sequence SEQ ID NO: 188, 196, 199, 202, 205, 210, 216, or 248, 282, 284, or 286, if first the heterodimeric region contains a cavity Fc chain, or a cavity Fc chain containing the sequence SEQ ID NO: 189, 197, 200, 203, 206, 211, 217, 220, 249, 283, 285 or 287, if such first heterodimeric the region contains an Fc chain with a knob (Table 7).

Table 9 shows the nucleic acid sequences of the first polypeptide chain and the second polypeptide chain of bispecific antibodies with various anti-GUCY2c m CD3 antibodies presented herein. SEQ ID NOs: 246 and 247 set forth the corresponding nucleotide sequences of the invention.

Table 9

First polypeptide chain GUCY2C-1608 (SEQ ID NO: 246) gacatccagctgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgcagagccagtgaaagtgttgattattatggcagtagtttattgcagtggtatcagcagaaaccagggaaagcccctaagctcctgatctatgctgcatccaaactagcttctggggtcccatcaaggttcagtggcagtggatct gggacagatttcactctcaccatcagcagtctgcaacctgaagattttgcaacttactactgtcagcaaactcggaaagcgtatacgtttggccaggggaccaagctggagatcaaaggtggaggtagcgggggcggcggggaagttcaactcgttgagtctggcgggggattggttcaacccggtggaagccttagattgtcatgtgcc gcctccggctttacatttagcgactattacatgacctgggtgagacaagctccaggcaaaggacttgaatgggtggcctttatcagaaatcaggcccgcggctacacaagcgaccataatccctccgtgaaaggaagatttaccatcagccgggacaatgctaaaaattcactttaccttcaaatgaactctcttagagccgaggacaccgccgtatactactg cgcaagagatagaccaagttattacgtcctggattactggggccagggaacaaccgtcaccgtgtcttctggatgcccaccgtgcccagcacctgaagccgctggggcaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtca agttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgaga accacaggtgtgcaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgtggtgcctggtcaaaggcttctatcccagcgacatcgccgtggagtggggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctatagcaagctcaccgtggacaagagcagg tggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtcccccggaaag Second polypeptide chain GUCY2C-1608 (SEQ ID NO: 247) gacatccagatgacccagtccccctcttctctgtctgcctctgtgggcgacagagtgaccatcacctgcacaagctcacagtcactgtttaatgtccgcagccagaaaaactatcttgcgtggtatcagcagaagcctggcaaggctcccaagctgctgatctactgggccagtacacgagaatccggcgtgccttccagattct ccggctctggctctggcaccgatttcaccctgaccatctcctccctccagcctgaggatttcgccacctactactgcaaacagtcttacgaccttttcacttttggcggcggaacaaaggtggagatcaagggcggaggtggatctggcggcggaggcgaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctga gactctcctgtgcagcctctggcttcaccttcagcagctactggatgcactgggtccgccaggctccagggaaggggctggagtggattggagagattaaacctagcaacgaacttactaacgtccatgaaaagttcaaggaccgattcaccatctccgtggacaaggccaagaactcagcctatctgcaaatgaacagcctgagagccgaggacacggctgt gtattactgtacaagaacgattacgacgacggagggatactggttcttcgatgtctggggccaagggacactggtcaccgtctcttcaggatgtccaccgtgcccagcacctgaagccgctggggcaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgag ccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaag ccaaagggcagccccgagaaccacaggtgtacaccctgcccccatgccgggaggagatgaccaagaaccaggtcagcctgtcctgcgcggtcaaaggcttctatcccagcgacatcgccgtggagtggggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctcgttagcaagct caccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtcccccggaaag

Effector null mutations in CH2-CH3 human IgG1

The human IgG1 Fc chain was modified to introduce mutations L234A, L235A and G237A (SEQ ID NO: 187, EU numbering) using standard primer-directed PCR mutagenesis to reduce effector function due to FcγRIII binding, resulting in null effector function phenotype (Canfield et al., J. Exp. Med (1991) 173:1483-1491; Shields et al., J. Biol. Chem. (2001) 276:6591-604).

Peak-to-trench mutations in human IgG1 CH2-CH3

Knobs-to-valves is an effective design strategy known in the art for designing antibody heavy chain homodimers for heterodimerization. In this approach, the overhang variant was generated by replacing a small amino acid with a larger amino acid on one strand of the IgG1 Fc chain, eg Y349C and T366W (EU numbering). The “knob” was designed to insert into a “cavity” in the CH3 domain of the complementary strand of the Fc chain, obtained by replacing a large residue with a smaller residue, for example, S354C, T366S, L368A and Y407V (numbered according to the EU index).

In some embodiments, complementary mutations are introduced to achieve heterodimerization of the resulting Fc chains such that each Fc chain will carry a set of mutations, Y349C and T366W in the case of a knob Fc chain (SEQ ID NO: 188), or S354C, T366S . domain, i.e. Preferably, the bispecific antibody contains one knob Fc chain linked to one knob Fc chain.

Table 10

Fc chain overhang mutations (SEQ ID NO: 188) APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPG Fc chain dimple mutations (SEQ ID NO: 189) APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG

The anti-CD3-GUCY2c bispecific antibodies of the present invention are stable against aggregation in thermal stability studies and are potent anti-CD3 and human GUCY2c bispecific antibody-Fc fusion proteins. The knob-to-valve Fc domain allows for improved expression in CHO cells and improved purification resulting in high purity of the desired heterodimer. Mutations engineered in the Fc domain abrogate FcγR binding, thereby potentially avoiding ADCC-mediated T cell exhaustion. In addition, the inclusion of an Fc domain in a bispecific antibody increases the stability of the molecules, analyzed by differential scanning calorimetry (DSC).

A bispecific antibody can be designed to contain at least one cysteine residue that can react with a corresponding cysteine residue on another polypeptide chain of the invention to form an interchain disulfide bond. Interchain disulfide bonds can serve to stabilize the bispecific antibody, improving expression and recovery in recombinant systems, resulting in a stable and uniform composition, as well as improving the stability of the isolated and/or purified product in vivo . The cysteine residue or residues can be inserted as a single amino acid or as part of a larger amino acid sequence, such as a hinge region, in any part of the polypeptide chain. In a particular aspect, the at least one cysteine residue is designed to be located at the C-terminus of the polypeptide chain.

In one aspect, the present invention provides a bispecific antibody that specifically binds to GUCY2c and competes for binding with the bispecific antibody provided herein. In some such embodiments, the bispecific antibody specifically binds to an epitope of GUCY2c.

In one aspect, the present invention provides a bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain, and wherein: (a) the first polypeptide chain contains the following regions in the following order in the direction from N- end to C-terminus: VL anti-GUCY2c antibody (VL anti-GUCY2c) (SEQ ID NO: 147) - linker 1 (SEQ ID NO: 190) - VH anti-CD3 antibody (VH anti-CD3) (SEQ ID NO: 9) - linker 3 (SEQ ID NO: 192) - first heterodimerization promoting domain (SEQ ID NO: 188); and (b) the second polypeptide chain contains the following regions in the following order in the direction from N-terminus to C-terminus: anti-CD3 VL (anti-CD3 VL) (SEQ ID NO: 90) - linker 2 (SEQ ID NO: 191) - VH antibodies against GUCY2c (VH against GUCY2c) (SEQ ID NO: 73) - linker 3 (SEQ ID NO: 192) - second heterodimerization promoting domain (SEQ ID NO: 189).

In another aspect, the invention provides a bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain, and where: (a) the first polypeptide chain contains the following regions in the following order in the direction from N-terminus to C-terminus: VL anti-GUCY2c antibodies (VL anti-GUCY2c) (SEQ ID NO: 147) - linker 1 (SEQ ID NO: 190) - VH anti-CD3 antibodies (VH anti-CD3) (SEQ ID NO: 9) - linker 3 (SEQ ID NO: 192) - first heterodimerization promoting domain (SEQ ID NO: 188); and (b) the second polypeptide chain contains the following regions in the following order in the direction from N-terminus to C-terminus: anti-CD3 VL (anti-CD3 VL) (SEQ ID NO: 90) - linker 2 (SEQ ID NO: 191) -VH antibodies against GUCY2c (VH against GUCY2c) (SEQ ID NO: 73) - linker 3 (SEQ ID NO: 192) - second heterodimerization promoting domain (SEQ ID NO: 189); where VL against GUCY2c and VH against GUCY2c form a domain that specifically binds to GUCY2c; and VL anti-CD3 and VH anti-CD3 form a domain that specifically binds to CD3; wherein the linker 3 of the first polypeptide chain and the linker 3 of the second polypeptide chain are covalently linked to each other by two disulfide bonds; wherein each of the first heterodimerization promoting domain and the second heterodimerization promoting domain comprises a CH2 domain and a CH3 domain; wherein the CH3 domain of the first heterodimerization promoting domain forms a protrusion, and the CH3 domain of the second heterodimerization promoting domain forms a recess; wherein at least one disulfide bond is formed between the CH3 domain of the first heterodimerization promoting domain and the CH3 domain of the second heterodimerization promoting domain.

In some such embodiments, the present invention provides a bispecific antibody that specifically binds to a GUCY2c epitope and a CD3 epitope.

In some embodiments, the present invention provides a bispecific antibody further comprising a human or humanized VH scaffold and a human or humanized VL scaffold. In some such embodiments, the bispecific antibody is a humanized antibody. In specific embodiments, the VH framework contains the sequence SEQ ID NO: 5, 6, 7, 8, 15, 16, 17, 18, 23, 24, 25, 30, 31, 32, 37, 38, 39, 40, 45, 46, 47, 49, 50, 51, 54, 55, 56, 58, 59, 61 or 63; and/or the VL framework contains the sequence SEQ ID NO: 80, 81, 82, 83, 86, 87, 88, 89, 96, 97, 98, 99, 103, 110, 111, 116, 117, 118, 122, 123 , 124, 126, 127, 128, 130, 131, 132, 133, 135, 139 or 155.

In one aspect, the bispecific antibody of the present invention specifically binds to GUCY2c and CD3, where the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain; where the first polypeptide chain is obtained using an expression vector with ATCC registration number PTA-124944; and the second polypeptide chain is obtained using an expression vector with ATCC registration number PTA-124943.

In one aspect, the present invention provides a bispecific antibody capable of specifically binding to GUCY2c and CD3, comprising a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain comprises the sequence SEQ ID NO: 216 and the second polypeptide chain comprises the sequence SEQ ID NO: 216. 220.

In some such embodiments, the present invention provides a bispecific antibody wherein the first and second polypeptide chains are covalently linked to each other by at least one disulfide bond.

The present invention further relates to a pharmaceutical composition containing a therapeutically effective amount of an antibody or bispecific antibody provided herein and a pharmaceutically acceptable carrier.

The present invention relates to methods and compositions for the treatment, prevention or control of cancer in an individual, comprising administering to the individual a therapeutically effective amount of an antibody or bispecific antibodies constructed according to the invention, wherein the molecule further binds to a cancer antigen. The molecules of the invention may be particularly suitable for preventing, inhibiting, reducing the growth and/or regression of primary tumors and metastasis of malignant cells. Although not limited to any particular mechanism of action, the molecules of the invention may mediate effector function, which may result in tumor clearance, tumor shrinkage, or a combination thereof.

The antibodies (eg, anti-GUCY2c or anti-CD3-GUCY2c bispecific antibodies) of the present invention can be used in a variety of ways, including, but not limited to, therapeutic methods and diagnostic methods.

In one aspect, the present invention provides a method for treating a condition associated with GUCY2c expression in an individual. In some embodiments, a method of treating a condition associated with GUCY2c expression in an individual comprises administering to the individual in need thereof an effective amount of a composition (eg, a pharmaceutical composition) containing anti-GUCY2c antibodies or anti-GUCY2c bispecific antibodies provided herein. Conditions associated with GUCY2c expression include, but are not limited to, abnormal GUCY2c expression, altered or abnormal GUCY2c expression, GUCY2c-expressing cancer cells, and a proliferative disorder (eg, cancer).

Thus, the invention relates to methods of treating a cancer characterized by the GUCY2c antigen by constructing a bispecific antibody for immunospecific recognition of the GUCY2c antigen and the CD3 antigen on T cells. Bispecific antibodies constructed according to the invention can be used for the prevention or treatment of malignant neoplasms, because they have cytotoxic activity due to their anti-CD3 antibody-induced activated killer T cells.

In a specific embodiment, the present invention provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient an anti-GUCY2c antibody as provided herein. The present invention also provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient a bispecific antibody of the present invention.

The present invention further provides a method of treating a GUCY2c-associated disorder in a patient in need thereof, comprising administering to the patient a pharmaceutical composition containing an anti-GUCY2c antibody or a bispecific antibody as provided herein. In some such embodiments, the GUCY2c-associated disorder is a malignancy. In specific embodiments, the cancer is breast, ovarian, thyroid, prostate, cervical, lung (including, but not limited to, non-small cell lung cancer and small cell lung cancer), bladder, endometrial, head and neck, testicular or glioblastoma. In some embodiments, the cancer is a digestive system cancer selected from the group consisting of cancer of the esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile duct, and pancreas. In specific embodiments, the therapy activates a cytolytic T cell response.

In one aspect, the present invention relates to an antibody, bispecific antibody or pharmaceutical composition provided herein for use in therapy. In a specific embodiment, the invention also provides an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody for use in a method of treating cancer as defined herein.

The present invention further relates to an antibody or bispecific antibody provided herein for use in the manufacture of a medicament for use in therapy. In some embodiments, the therapy is a treatment for a GUCY2c-associated disorder. In specific embodiments, the GUCY2c-associated disorder is a malignancy. In some embodiments, the cancer is selected from the group consisting of breast, ovarian, thyroid, prostate, cervical, lung (including, but not limited to, non-small cell lung cancer and small cell lung cancer), bladder, endometrial, head and neck, testicular or glioblastoma. In some embodiments, the cancer is a digestive system cancer selected from the group consisting of cancer of the esophagus, stomach, small intestine, colon, rectum, anal canal, liver, gallbladder, appendix, bile duct, and pancreas. In specific embodiments, the therapy activates a cytolytic T cell response.

In one aspect, the present invention relates to a polynucleotide encoding an antibody or bispecific antibody as described herein. In another embodiment, the present invention relates to a vector containing the polynucleotides presented in the present description. In yet another embodiment, the present invention relates to a host cell transformed or transfected using the vectors presented herein. In some such embodiments, the host cell recombinantly produces an antibody or bispecific antibody provided herein. In specific embodiments, the host cell is selected from the group consisting of bacterial cell lines, mammalian cell lines, insect cell lines, and yeast cell lines. In a specific embodiment, the mammalian cell line is a CHO cell line. In one embodiment, the antibody or bispecific antibody is produced using a cell-free in vitro protein synthesis system.

In one aspect, the present invention provides a method of treating cancer in a subject in need thereof, comprising a) obtaining a bispecific antibody as provided herein, and b) administering said bispecific antibody to said patient. In some embodiments, the present invention relates to a method of treating a condition associated with malignant cells expressing a tumor antigen in an individual, comprising a) obtaining an antibody provided herein, and b) administering to an individual in need thereof an effective amount of a pharmaceutical composition containing the antibody presented in this description. Non-limiting examples of cancers to be treated include esophageal cancer, stomach cancer, small intestinal cancer, colon cancer, rectal cancer, anal cancer, liver cancer, gallbladder cancer, appendix cancer, bile duct cancer and pancreatic cancer. Thus, the compositions and methods presented herein can be used for the treatment, imaging, detection and vaccination against the above primary and metastatic malignancies.

In some embodiments, the present invention provides a method of inhibiting the growth or progression of a tumor in an individual having cancer cells expressing GUCY2c, comprising administering to the individual in need thereof an effective amount of a composition comprising anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies as provided herein . In other embodiments, the present invention provides a method of inhibiting metastasis of cells expressing GUCY2c in an individual, comprising administering to the individual in need thereof an effective amount of a composition comprising anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies as provided herein. In other embodiments, the present invention provides a method of inducing tumor regression in malignant cells in an individual, comprising administering to the individual in need thereof an effective amount of a composition comprising anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies as provided herein.

In a specific aspect, the anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibody of the invention inhibit or reduce the growth of malignant cells by at least 99%, at least 95%, at least 90%, at least 85%, by at least 80%, at least 75%, at least 70%, at least 60%, at least 50%, at least 45%, at least 40%, at least at least 45%, at least 35%, at least 30%, at least 25%, at least 20%, or at least 10% relative to the growth of malignant cells in the absence of the antibody or bispecific antibody invention.

In a particular aspect, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies kill cells or inhibit or reduce the growth of malignant cells by at least 5%, at least 10%, at least 20%, at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least 45%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100% better than in the absence of the antibody or bispecific antibodies invention.

In one embodiment, the present invention provides a method of treating an autoimmune disorder in an individual, comprising administering to the individual in need thereof an effective amount of a composition comprising anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies as provided herein.

As used herein, autoimmune disorders include, but are not limited to, inflammatory bowel disease, systemic lupus erythematosus, diabetes (type I), celiac disease, agammaglobulinemia, autoimmune enteropathy, autoimmune hepatitis, Crohn's disease, gastrointestinal pemphigoid, polymyositis, transverse myelitis, ulcerative colitis and vasculitis.

In one aspect, the present invention provides an effective amount of a composition (e.g., a pharmaceutical composition) containing antibodies (e.g., anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein for treating a condition (e.g., cancer) associated with GUCY2c expression in an individual in need thereof. In one embodiment, the present invention provides an effective amount of a composition (eg, a pharmaceutical composition) containing antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies), as provided herein, for inhibiting the growth or progression of a tumor in an individual having malignant cells expressing GUCY2c. In some embodiments, the present invention provides an effective amount of a composition (e.g., a pharmaceutical composition) containing antibodies (e.g., anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) as provided herein for inhibiting the metastasis of cancer cells expressing GUCY2c, from the individual in need. In some embodiments, the present invention provides an effective amount of a composition (eg, a pharmaceutical composition) containing antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein.

In another aspect, the invention provides antibodies (eg, GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein for use in treating a condition (eg, cancer) associated with GUCY2c expression in an individual in need thereof. In some embodiments, the present invention provides antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein for inhibiting the growth or progression of a tumor in an individual having cancer cells expressing GUCY2c. In some embodiments, the present invention provides antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein for inhibiting the metastasis of GUCY2c-expressing cancer cells in an individual in need thereof. In some embodiments, the present invention provides antibodies (eg, GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein for inducing tumor regression in an individual having cancer cells expressing GUCY2c.

In another aspect, the invention relates to the use of antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein in the manufacture of a medicament for treating a condition (eg, cancer) associated with GUCY2c expression. In some embodiments, the present invention relates to the use of antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein in the manufacture of a medicament for inhibiting tumor growth or progression. In some embodiments, the present invention relates to the use of antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein in the manufacture of a medicament for inhibiting metastasis of cancer cells expressing GUCY2c. In some embodiments, the present invention relates to the use of antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein in the manufacture of a medicament for inducing tumor regression.

In another aspect, the present invention relates to a method for detecting, diagnosing and/or monitoring a condition associated with GUCY2c expression. For example, antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein can be labeled with a detectable moiety, such as an imaging agent and an enzyme-substrate label. The antibodies provided herein can also be used for in vivo diagnostic assays, such as in vivo imaging (eg, PET or SPECT), or as a staining reagent.

Thus, in addition to redirecting T cells to tumor-specific antigens, the anti-CD3-GUCY2c bispecific antibodies of the present invention can be used to deliver other diagnostic or therapeutic agents to cells expressing a tumor antigen on their surface. Thus, the bispecific antibody can be attached directly or indirectly, for example through a linker, to the drug so that it is delivered directly to cells bearing the tumor antigen. Therapeutics include, but are not limited to, nucleic acids, proteins, peptides, amino acids or derivatives, glycoproteins, radioactive isotopes, lipids, carbohydrates, or recombinant viruses. Nucleic acid-based diagnostic or therapeutic agents include, but are not limited to, antisense nucleic acids, derivatized oligonucleotides for covalent cross-linking to single or duplex DNA, and triplex-forming oligonucleotides.

Alternatively, diagnostic or therapeutic agents associated with the bispecific antibodies of the present invention may be an encapsulation system, such as a liposome or micelle, containing a therapeutic agent, such as a drug, nucleic acid (e.g., antisense nucleic acid), or other therapeutic agent, preferably protected from direct exposure to the bloodstream. Methods for preparing liposomes attached to antibodies are well known to those skilled in the art. See, for example, US Pat. No. 4,957,735; and Connor, et al., Pharm. Ther. 28:341-365 (1985).

In some embodiments, the methods provided herein further include the step of treating an individual using an additional form of therapy. In some embodiments, the additional form of therapy is additional antineoplastic therapy, including, but not limited to, chemotherapy, radiation therapy, surgery, hormonal therapy, and/or additional immunotherapy.

The present invention further includes administration of a molecule of the invention in combination with other therapeutic agents known to those skilled in the art for the treatment or prevention of cancer, including, but not limited to, standard and experimental chemotherapeutic agents, biological therapeutic agents, immunotherapy agents, radiation therapy, or surgical intervention. In some aspects, molecules of the invention may be administered in combination with a therapeutically or prophylactically effective amount of one or more agents, therapeutic antibodies, or other agents known to those skilled in the art for the treatment and/or prevention of cancer.

Thus, methods of treating cancer include administering to an individual in need thereof an effective amount of an antibody or bispecific antibody of the present invention in combination with a chemotherapeutic agent. Such combination treatments may be administered separately, sequentially, or simultaneously. Suitable chemotherapeutic agents include, by way of non-limiting examples, at least one additional agent such as bevacizumab, cetuximab, sirolimus, panitumumab, 5-fluorouracil (5-FU), capecitabine, tivozanib, irinotecan, oxaliplatin, cisplatin, trifluridine, tipiracil, leucovorin, gemcitabine and/or erlotinib hydrochloride.

Dosage amounts and frequencies of administration provided herein are included within the terms "therapeutically effective" and "prophylactically effective." The dosage and frequency further vary depending on factors specific to each patient, depending on the specific therapeutic or prophylactic agents administered, the severity and type of malignancy, the route of administration, and the age, weight, response and medical history of the patient. One skilled in the art can select appropriate treatment regimens by considering such factors and following, for example, the doses described in the literature and recommended in the Physician's Desk Reference (56th edition, 2002).

The antibodies or bispecific antibodies of the present invention may be in a pharmaceutical composition for administration formulated to be suitable for the selected route of administration and containing pharmaceutically acceptable diluent or excipients such as buffers, surfactants, preservatives, solubilizers, isotonic agents, stabilizing agents. means, media, etc. Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton Pa., ^18th ed., 1995 provides guidance on formulation methods generally known to those skilled in the art.

These pharmaceutical compositions can be administered by any means known in this field, by which, as a rule, the intended purpose of treating a malignant neoplasm is achieved. The route of administration may be parenteral, defined herein as referring to methods of administration including, but not limited to, transesophageal, intratumoral, transcolonoscopic, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous and intra-articular injection and infusion. The method of administration and dosage of the molecules of the invention (eg, anti-GUCY2c antibodies, anti-CD3-GUCY2c bispecific antibodies, pharmaceutical composition) depends on the type of disease being treated, if necessary, its stage, the antigen to be monitored, the type of concomitant treatment, if any. , frequency of treatment, nature of the desired effect, and the patient's weight, age, health, nutrition, and gender. Thus, the particular dosage regimen used may vary widely and thus may deviate from the dosage regimens described herein.

Various antibody formulations of the present invention (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) can be used for administration. In some embodiments, antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibody) may be administered alone. In some embodiments, the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) and the pharmaceutically acceptable excipient may be in different formulations. Pharmaceutically acceptable excipients are known in the art and are relatively inert substances that facilitate the administration of a pharmacologically effective substance. For example, the excipient may impart shape or consistency or act as a diluent. Suitable excipients include, but are not limited to, stabilizing agents, humectants and emulsifiers, osmolarity varying salts, encapsulating agents, buffers and skin penetration enhancers. Excipients, as well as formulations for parenteral and non-parenteral administration of the drug, are given in Remington, The Science and Practice of Pharmacy 21st Ed. Mack Publishing, 2005.

In some embodiments, these agents are formulated for administration by injection (eg, intraperitoneally, intravenously, subcutaneously, intramuscularly, etc.). Thus, these agents can be combined with pharmaceutically acceptable carriers such as saline, Ringer's solution, dextrose solution and the like. The specific dosage regimen, i.e. dose, timing and repetition will depend on the individual and the individual's medical history.

Antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) provided herein can be administered by any suitable route, including injection (eg, intraperitoneal, intravenous, subcutaneous, intramuscular, etc.). An antibody, such as a monoclonal antibody or a bispecific antibody, is also administered by inhalation as described herein. Generally, when administering an antibody of the present invention (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody), the dose depends on the organism being treated and the particular route of administration. In one embodiment, the dosage range of the antibody of the present invention will be from about 0.001 μg/kg body weight to about 20,000 μg/kg body weight. The term "body weight" is used when the patient is being treated. If isolated cells are treated, the term "body weight" as used herein refers to "total cell weight". The term "total body weight" can be used in relation to the treatment of the isolated cell and the treatment of the patient. All concentrations and treatment levels expressed herein as "body weight" or simply "kg" are also considered to cover similar concentrations of "total cell mass" and "total body weight". However, those skilled in the art will recognize the usefulness of various dosage ranges, for example, from 0.01 μg/kg body weight to 20,000 μg/kg body weight, from 0.02 μg/kg body weight to 15,000 μg/kg body weight, from 0 .03 mcg/kg body weight to 10,000 mcg/kg body weight, from 0.04 mcg/kg body weight to 5000 mcg/kg body weight, from 0.05 mcg/kg body weight to 2500 mcg/kg body weight, from 0.06 µg/kg body weight to 1000 µg/kg body weight, from 0.07 µg/kg body weight to 500 µg/kg body weight, from 0.08 µg/kg body weight to 400 µg/kg body weight, from 0.09 µg/kg body weight to 200 µg/kg body weight or from 0.1 µg/kg body weight to 100 µg/kg body weight. In addition, those skilled in the art will appreciate that various dose levels can be used, for example, 0.0001 µg/kg, 0.0002 µg/kg, 0.0003 µg/kg, 0.0004 µg/kg, 0.005 µg/kg, 0 .0007 µg/kg, 0.001 µg/kg, 0.1 µg/kg, 1.0 µg/kg, 1.5 µg/kg, 2.0 µg/kg, 5.0 µg/kg, 10.0 µg /kg, 15.0 mcg/kg, 30.0 mcg/kg, 50 mcg/kg, 75 mcg/kg, 80 mcg/kg, 90 mcg/kg, 100 mcg/kg, 120 mcg/kg, 140 mcg/ kg, 150 mcg/kg, 160 mcg/kg, 180 mcg/kg, 200 mcg/kg, 225 mcg/kg, 250 mcg/kg, 275 mcg/kg, 300 mcg/kg, 325 mcg/kg, 350 mcg/ kg, 375 mcg/kg, 400 mcg/kg, 450 mcg/kg, 500 mcg/kg, 550 mcg/kg, 600 mcg/kg, 700 mcg/kg, 750 mcg/kg, 800 mcg/kg, 900 mcg/ kg, 1 mcg/kg, 5 mcg/kg, 10 mcg/kg, 12 mcg/kg, 15 mg/kg, 20 mg/kg and/or 30 mg/kg. All of these doses are examples, and it is expected that any dose between these values can be used in the present invention. Any of the above dosage ranges or dosage levels may be used for the antibody of the present invention. For repeated administrations over several days or more, depending on the condition, treatment is continued until the desired suppression of symptoms occurs or until sufficient therapeutic levels are achieved, for example, to inhibit or delay tumor growth/progression or metastasis of malignant cells.

Other dosing regimens may also be used depending on the pharmacokinetic disintegration profile that the practitioner wishes to achieve. In one embodiment, the antibody of the present invention is administered at an increased initial dose, followed by a higher and/or continuous, substantially constant dose. In some embodiments, administration is provided from one to four times per week. In other embodiments, administration is provided once a month, or once every two months, or once every three months. The progress of this therapy is easily monitored using conventional methods and assays. The dosing regimen (including the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody)) can be varied over time.

For purposes of the present invention, the appropriate dose of the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) will depend on the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) or compositions thereof used, the type and severity of symptoms treated, if the drug is administered for therapeutic purposes, previous therapy, the patient's clinical history and response to the drug, the rate of elimination of the administered drug from the patient and the decision of the treating physician. Typically, the clinician will administer the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) until the dose is achieved to achieve the desired outcome. The dose and/or frequency may be varied during treatment. Dose determination will generally take into account empirical factors such as half-life. For example, antibodies compatible with the human immune system, such as humanized antibodies or fully human antibodies, can be used to prolong the half-life of the antibody and to prevent the antibody from being attacked by the host immune system. The frequency of administration can be determined and adjusted during therapy and is typically, but not necessarily, based on treatment and/or suppression and/or improvement and/or delay of symptoms, e.g., inhibition or delay of tumor growth, etc. d. Alternatively, sustained release antibody formulations (eg, GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibody) may be suitable. Various formulations and devices for achieving sustained release are known in the art.

In one embodiment, dosages of the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) may be determined empirically in individuals subjected to one or more administrations of the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody). Individuals are administered increasing doses of an antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody). Disease scores can be monitored to evaluate effectiveness.

In some embodiments, administration of an antibody (e.g., anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) results in at least one effect selected from the group consisting of tumor growth inhibition, tumor regression, tumor size reduction, tumor cell count reduction, delayed tumor growth, abscopal effect, inhibition of tumor metastasis, reduction of metastatic lesions over time, reduction in the use of chemotherapeutic or cytotoxic agents, reduction of tumor burden, improved progression-free survival, improved overall survival, complete response, partial response and stable disease.

Administration of an antibody (e.g., anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) in accordance with the method of the present invention may be continuous or intermittent, depending, for example, on the physiological state of the recipient, whether the purpose of administration is therapeutic or prophylactic, and other factors , known to specialists in this field. Administration of the antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) may be substantially continuous over a preselected period of time or may be a series of divided doses.

In some embodiments, multiple antibodies may be present (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody). At least one, at least two, at least three, at least four, at least five, or more different antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) may be present. In general, antibodies (eg, anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) may have complementary activities without negatively affecting each other. For example, one or more of the following antibodies may be used: a first anti-GUCY2c or CD3 antibody directed against one epitope on GUCY2c or CD3, and a second anti-GUCY2c or CD3 antibody directed against another epitope on GUCY2c or CD3.

The therapeutic antibody compositions (e.g., anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies) used in the present invention are prepared for storage by mixing the antibody having the desired purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington, The Science and Practice of Pharmacy 21st Ed. Mack Publishing, 2005), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients or stabilizers are non-toxic to recipients at the doses and concentrations used, and may contain buffers such as phosphate, citrate and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkylparabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; complex compounds with metals (for example, Zn-protein complexes); and/or non-ionic surfactants such as TWEEN ^™ , PLURONIC ^™ or polyethylene glycol (PEG).

Liposomes containing an antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) are prepared by methods known in the art, as described in Epstein, et al., Proc. Natl. Acad. Sci. USA 82:3688, 1985; Hwang, et al., Proc. Natl Acad. Sci. USA 77:4030, 1980; and US Pat. Nos. 4,485,045 and 4,544,545. Extended circulation time liposomes are described in US Pat. No. 5,013,556. Particularly suitable liposomes can be prepared by a reverse phase evaporation process using a lipid composition containing phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE) . Liposomes are extruded through filters with specific pore sizes to obtain liposomes with the desired diameter.

Active ingredients can also be enclosed in microcapsules obtained, for example, by coacervation methods or by interphase polymerization, for example, hydroxymethylcellulose or gelatin microcapsules and polymethyl methacrylate microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions. Such methods are described in Remington, The Science and Practice of Pharmacy 21st Ed. Mack Publishing, 2005.

Slow-release formulations can be obtained. Suitable examples of sustained release formulations include semi-permeable matrices of solid hydrophobic polymers containing the antibody, where the matrices are in the form of shaped articles, for example films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (eg, poly(2-hydroxyethyl methacrylate) or polyvinyl alcohol), polylactides (US Patent No. 3,773,919), copolymers of L-glutamic acid and 7-ethyl-L-glutamate, non-degradable ethylene vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT ^TM (injectable microspheres consisting of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate and poly-D-(-)-3-hydroxybutyric acid.

Formulations intended for use for in vivo administration must be sterile. This is easily achieved, for example, by filtration through sterilization filtration membranes. Therapeutic antibody compositions (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) are typically placed in a container having a sterile entry port, such as an intravenous solution bag or vial with a stopper pierced by a hypodermic needle.

The compositions of the present invention may be in unit dosage forms, such as tablets, pills, capsules, powders, granules, solutions or suspensions or suppositories, for oral, parenteral or rectal administration, or administration by inhalation or insufflation.

To prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical carrier, for example, conventional tabletting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium or gum phosphate, and others. pharmaceutical diluents, for example, water, to obtain a solid composition before giving it a dosage form containing a homogeneous mixture of a compound of the present invention or a non-toxic pharmaceutically acceptable salt thereof. By referring to these compositions as homogeneous prior to dosage forming, it is meant that the active ingredient is uniformly dispersed throughout the composition so that the composition can be readily divided into equally effective unit dosage forms such as tablets, pills and capsules. This solid composition is then divided into unit dosage forms of the type described above, containing from 0.1 to about 500 mg of the active ingredient of the present invention, before being formulated into a dosage form. Tablets or pills of the new composition can be coated or otherwise extemporaneously prepared to obtain a dosage form that provides the advantage of prolonged release. For example, a tablet or pill may contain an internal dose component and an external dose component, the latter of which is in the form of a coating around the former. The two components can be separated by an enteric layer designed to resist gastric degradation and allow the internal component to enter the duodenum or have a sustained release. Various materials can be used for such enteric layers or coatings, including a number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate.

Suitable surfactants include, in particular, non-ionic agents such as polyoxyethylene sorbitans (eg Tween ^TM 20, 40, 60, 80 or 85) and other sorbitans (eg Span ^TM 20, 40, 60, 80 or 85). Surfactant compositions will typically contain from 0.05 to 5% surfactant and may contain from 0.1 to 2.5%. It should be understood that, if necessary, other ingredients, such as mannitol or other pharmaceutically acceptable carriers, can be added.

Suitable emulsions can be prepared using commercially available fat emulsions such as Intralipid ^™ , Liposyn ^™ , Infonutrol ^™ , Lipofundin ^™ and Lipiphysan ^™ . The active ingredient may be dissolved in a premixed emulsion composition or, alternatively, it may be dissolved in an oil (eg, soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or almond oil) and an emulsion obtained after mixing with a phospholipid (eg, egg phospholipids, soy phospholipids or soy lecithin) and water. It should be understood that other ingredients, such as glycerin or glucose, can be added to adjust the tonicity of the emulsion. Suitable emulsions will typically contain up to 20% oil, for example 5 to 20%. The fat emulsion may contain fat droplets from 0.1 to 1.0 µm, in particular from 0.1 to 0.5 µm, and have a pH in the range from 5.5 to 8.0.

Emulsion compositions can be compositions prepared by mixing an antibody (eg, anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) with Intralipid ^™ or its components (soybean oil, egg phospholipids, glycerol and water).

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. The compositions in preferably sterile pharmaceutically acceptable solvents can be nebulized using gases. Nebulizable solutions can be inhaled directly from a nebulizing device, or the nebulizing device can be attached to a face mask, tent, or positive intermittent pressure breathing apparatus. The compositions in the form of a solution, suspension or powder can be administered, preferably orally or nasally using a device through which the composition is administered in an appropriate manner.

Compositions within the scope of the invention include all compositions wherein the antibody (anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody) is present in an amount effective to achieve the desired medical effect in the treatment of cancer. Although individual needs may vary from one patient to another, determining the optimal ranges of effective amounts of all components is within the skill of one skilled in the art.

Examples of such compositions, as well as methods of composition, are also described above and below. In some embodiments, the composition contains one or more antibodies (eg, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody). In some embodiments, the anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody is a human antibody, a humanized antibody, or a chimeric antibody. In some embodiments, the anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody comprises a constant region capable of triggering a desired immune response, such as antibody-mediated lysis or ADCC. In other embodiments, the anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody comprises a constant region that does not trigger an unwanted immune response, such as antibody-mediated lysis or ADCC.

It should be understood that the compositions may contain multiple antibodies (for example, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody), for example, a mixture of anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies recognizing different epitopes of GUCY2c or CD3 and GUCY2c. Other examples of compositions contain multiple anti-GUCY2c antibodies or anti-CD3-GUCY2c bispecific antibodies recognizing the same epitopes, or different types of anti-GUCY2c antibodies, anti-CD3-GUCY2c bispecific antibodies, or antibodies that bind to different GUCY2c epitopes (e.g., human GUCY2cA) or CD3 and GUCY2c (human CD3 and GUCY2c).

In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody may be administered in combination with the administration of one or more additional therapeutic agents. These include, but are not limited to, administration of a biotherapeutic agent and/or a chemotherapeutic agent, such as, but not limited to, a vaccine, CAR-T cell therapy, radiation therapy, cytokine therapy, an anti-CD3 bispecific antibody, an inhibitor of other immunosuppressive pathways, angiogenesis inhibitor, T cell activator, metabolic pathway inhibitor, mTOR inhibitor, adenosine pathway inhibitor, tyrosine kinase inhibitor including, but not limited to, Inlyta, ALK inhibitors and sunitinib, BRAF inhibitor, epigenetic modifier, IDO1 inhibitor, JAK inhibitor, STAT inhibitor, cyclin-dependent kinase inhibitor, biotherapeutic (including, but not limited to, antibodies against VEGF, VEGFR, EGFR, Her2/neu, other growth factor receptors, CD40, CD-40L, CTLA-4, OX-40, 4-1BB , TIGIT and ICOS), immunogenic agent (eg, attenuated malignant cells, tumor antigens, antigen presenting cells such as tumor antigen or nucleic acid-injected dendritic cells, immunostimulatory cytokines (eg, IL-2, IFNα2, GM-CSF) and cells transfected with genes encoding immunostimulatory cytokines such as, but not limited to, GM-CSF).

Examples of biotherapeutics include therapeutic antibodies, immunomodulatory agents, and therapeutic immune cells.

Therapeutic antibodies may have specificity for various antigens. For example, therapeutic antibodies can be directed against a tumor-associated antigen such that binding of the antibody to the antigen promotes the death of the cell expressing the antigen. In another example, therapeutic antibodies can be directed against an antigen (eg, PD-1) on an immune cell such that binding of the antibody prevents down-regulation of the activity of the antigen-expressing cell (and thus promotes the activity of the antigen-expressing cell). In some cases, a therapeutic antibody may function through a variety of different mechanisms (eg, it may i) promote the death of a cell expressing the antigen, and ii) prevent the antigen from inducing downregulation of the activity of immune cells in contact with the cell expressing the antigen).

Therapeutic antibodies can be directed against, for example, the antigens listed below. For some antigens, examples of antibodies against the antigen are also included below (in parentheses/parentheses after the antigen). The following antigens may also be referred to herein as "target antigens" or the like. Target antigens for therapeutic antibodies herein include, for example: 4-1BB (eg, utomilumab); 5T4; A33; alpha-folate receptor 1 (eg, mirvetuximab soravtansine); Alk-1; BCMA [eg, PF-06863135 (see US Patent No. 9969809)]; BTN1A1 (for example, see WO2018222689); CA-125 (eg, abagovomab); carbonic anhydrase IX; CCR2; CCR4 (eg, mogamulizumab); CCR5 (eg leronlimab); CCR8; CD3 [eg, blinatumomab (anti-CD3/CD19 bispecific antibody), PF-06671008 (anti-CD3/P-cadherin bispecific antibody), PF-06863135 (anti-CD3/BCMA bispecific antibody), CD19 (eg, blinatumomab, MOR208); CD20 (eg ibritumomab tiuxetan, obinutuzumab, ofatumumab, rituximab, ublituximab); CD22 (inotuzumab ozogamicin, moxetumomab pasudotox); CD25; CD28; CD30 (eg, brentuximab vedotin); CD33 (eg gemtuzumab ozogamicin); CD38 (eg, daratumumab, isatuximab), CD40; CD-40L; CD44v6; CD47; CD52 (eg, alemtuzumab); CD63; CD79 (eg, polatuzumab vedotin); CD80; CD123; CD276/B7-H3 (eg, omburtamab); CDH17; CEA; ClhCG; CTLA-4 (eg, ipilimumab, tremelimumab), CXCR4; desmoglein 4; DLL3 (eg, rovalpituzumab tesirin); DLL4; E-cadherin; EDA; EDB; EFNA4; EGFR (eg, cetuximab, depatuxizumab mafodotin, necitumumab, panitumumab); EGFRvIII; endosialin; EpCAM (eg, oportuzumab monatox); FAP; fetal acetylcholine receptor; FLT3 (eg see WO2018/220584); GD2 (eg dinutuximab, 3F8); GD3; GITR; GloboH; GM1; GM2; GUCY2C (eg PF-07062119); HER2/neu [eg, margetuximab, pertuzumab, trastuzumab; ado-trastuzumab emtansine, trastuzumab duocarmazine, PF-06804103 (see US8828401)]; HER3; HER4; ICOS; IL-10; ITG-AvB6; LAG-3 (eg, relatlimab); Lewis Y antigen; LG; Ly-6; M-CSF [eg PD-0360324 (see US Patent No. 7326414)]; MCSP; mesothelin; MUC1; MUC2; MUC3; MUC4; MUC5AC; MUC5B; MUC7; MUC16; Notch1; Notch3; nectin-4 (eg, enfortumab vedotin); OX40 [eg, PF-04518600 (see US Patent No. 7960515)]; P-cadherin [eg PF-06671008 (see WO2016/001810)]; PCDHB2; PD-1 [eg, BCD-100, camrelizumab, cemiplimab, genolimzumab (CBT-501), MEDI0680, nivolumab, pembrolizumab, RN888 (see WO2016/092419), sintilimab, spartalizumab, STI-A1110, tislelizumab, TSR-042] ; PD-L1 (eg, atezolizumab, durvalumab, BMS-936559 (MDX-1105) or LY3300054); PDGFRA (eg, olaratumab); plasma cell antigen; poly-SA; PSCA; PSMA; PTK7 [eg, PF-06647020 (see US Patent No. 9409995)]; Ror1; SAS; SCRx6; SLAMF7 (eg, elotuzumab); SHH; SIRPa (eg ED9, Effi-DEM); STEAP; TGF-beta; TIGIT; TIM-3; TMPRSS3; TNF-alpha precursor; TROP-2 (eg, sacituzumab govitecan); TSPAN8; VEGF (eg, bevacizumab, brolucizumab); VEGFR1 (eg, ranibizumab); VEGFR2 (eg, ramucirumab, ranibizumab); Wue-1.

The therapeutic antibodies may be in any suitable format. For example, therapeutic antibodies can be in any format described elsewhere herein. In some embodiments, the therapeutic antibody may be a naked antibody. In some embodiments, the therapeutic antibody may be linked to a drug/agent (also known as an “antibody-drug conjugate” (ADC)). In some embodiments, a therapeutic antibody against a specific antigen may be included in a multispecific antibody (eg, a bispecific antibody).

In some embodiments, an antibody against an antigen can be conjugated to a drug/agent. Bound antibody-drug molecules are also referred to as “antibody-drug conjugates” (ADCs). Drugs/agents can be coupled to the antibody directly or indirectly through a linker. Most often, toxic drugs are combined with an antibody in such a way that the binding of the ADC to the corresponding antigen promotes the destruction of cells expressing the antigen. For example, ADCs that are coupled to toxic drugs are particularly suitable for targeting tumor-associated antigens to promote the killing of tumor cells expressing tumor-associated antigens. In other embodiments, the agents that can be coupled to the antibody may be, for example, an immunomodulatory agent (for example, to modulate the activity of immune cells in the vicinity of an ADC), an imaging agent (for example, to facilitate visualization of an ADC in an individual or in a biological sample of an individual), or an agent to increase the serum half-life or biological activity of an antibody.

Methods for conjugating a cytotoxic agent or other therapeutic agents with antibodies are described in various publications. For example, chemical modification can be carried out in antibodies through lysine side chain amines or cysteine sulfhydryl groups activated by reduction of interchain disulfide bonds to allow a conjugation reaction to occur. See, for example, Tanaka et al., FEBS Letters 579:2092-2096, 2005, and Gentle et al., Bioconjugate Chem. 15:658-663, 2004. Reactive cysteine residues designed at specific antibody sites for specific drug conjugation with a specific stoichiometry are also described. See, for example, Junutula et al., Nature Biotechnology, 26:925-932, 2008. Conjugation using a tag containing an acyl donor glutamine, or endogenous glutamine made reactive (i.e., with the ability to form a covalent bond as acyl donor) by constructing a polypeptide in the presence of a transglutaminase and an amine (eg, a cytotoxic agent containing or attached to a reactive amine), is also described in international applications WO2012/059882 and WO2015015448. In some embodiments, an ADC may have any of the features or characteristics of an ADC presented in WO2016166629, thereby being incorporated by reference for all purposes.

Drugs/agents that can be coupled to an antibody in ADC format may include, for example, cytotoxic agents, immunomodulatory agents, imaging agents, therapeutic proteins, biopolymers, or oligonucleotides.

Examples of cytotoxic agents that can be included in the ADC include anthracycline, auristatin, dolastatin, carrier, duokocarmitsin, dimer pyrrolobenzodiazepine, dimer indolino-Benzodiazepinp, yendin, geldanamycin, mayanzin, puromycin, dachshunds, coaline barvinovinka, camptothecine, tubulizin, tubulizin , hemiasterlin, Playsostatin, pladienolide and their stereoisomers, isostereomers, analogues or derivatives.

Examples of immunomodulatory agents that may be included in an ADC include ganciclovir, etanercept, tacrolimus, sirolimus, voclosporin, cyclosporine, rapamycin, cyclophosphamide, azathioprine, mycophenolate mofetil, methotrexate, glucocorticoid and its analogues, cytokines, stem cell growth factors, lymphotoxins, necrosis factor tumors (TNF), hematopoietic factors, interleukins (for example, interleukin-1 (IL-1), IL-2, IL-3, IL-6, IL-10, IL-12, IL-18 and IL-21), colony-stimulating factors (eg, granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF)), interferons (eg, interferon-alpha, -beta, and gamma), stem cell growth factor, designated "factor S 1", erythropoietin and thrombopoietin or a combination thereof.

Examples of imaging agents that can be included in the ADC include fluorescein, rhodamine, lanthanide-activated phosphorus and derivatives thereof, or a radioactive isotope coupled to a chelator. Non-limiting examples of fluorophores include fluorescein isothiocyanate (FITC) (e.g., 5-FITC), fluorescein amidite (FAM) (e.g., 5-FAM), eosin, carboxyfluorescein, erythrosine, Alexa Fluor ^™ (e.g., Alexa 350, 405, 430, 488, 500, 514, 532, 546, 555, 568, 594, 610, 633, 647, 660, 680, 700 or 750), carboxytetramethylrhodamine (TAMRA) (e.g. 5,-TAMRA), tetramethylrhodamine (TMR) and sulforhodamine (SR ) (for example, SR101). Non-limiting examples of chelators include 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), 1,4,7-triazacyclononane, 1-glutaric acid-4,7-acetic acid (deferoxamine), diethylenetriaminepentaacetic acid (DTPA) and 1,2-bis(o-aminophenoxy)ethane-N,N,N ',N'-tetraacetic acid (BAPTA).

Examples of therapeutic proteins that can be included in an ADC include a toxin, a hormone, an enzyme, and a growth factor.

Examples of biocompatible polymers that can be included in ADCs include water-soluble polymers such as polyethylene glycol (PEG) or derivatives thereof and zwitterion-containing biocompatible polymers (eg, phosphorylcholine-containing polymer).

Examples of biocompatible polymers that can be included in ADCs include antisense oligonucleotides.

In some embodiments, an antibody against an antigen provided herein can be included in a bispecific antibody molecule. Bispecific antibodies are monoclonal antibodies that have binding specificity for at least two different antigens.

Immunomodulatory agents include various types of molecules that can stimulate an immune response in an individual, such as pattern recognition receptor (PRR) agonists, immunostimulatory cytokines, and antitumor vaccines.

Pattern recognition receptors (PRRs) are receptors expressed by cells of the immune system that recognize various molecules associated with pathogens and/or cell damage or death. PRRs are involved in the innate immune response and the adaptive immune response. PRR agonists can be used to stimulate an immune response in an individual. There are many classes of PRR molecules, including Toll-like receptors (TLRs), RIG-I-like receptors (RLRs), nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), C-type lectin receptors (CLRs), and protein -stimulator of interferon genes (STING).

The terms "TLR" and "Toll-like receptor" refer to any Toll-like receptor. Toll-like receptors are receptors involved in the activation of immune responses. TLRs recognize, for example, pathogen-associated molecular patterns (PAMPs) expressed in microorganisms, as well as endogenous damage-associated molecular patterns (DAMPs) released from dead or dying cells.

Molecules that activate TLRs (and thus activate immune responses) are referred to herein as “TLR agonists.” TLR agonists may include, for example, small molecules (eg, an organic molecule having a molecular weight of up to about 1000 Da) as well as large molecules (eg, oligonucleotides and proteins). Some TLR agonists are specific for one type of TLR (eg, TLR3 or TLR9), while some TLR agonists activate two or more types of TLR (eg, both TLR7 and TLR8).

Examples of TLR agonists provided herein include TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 and TLR9 agonists.

Examples of small molecule TLR agonists include those described, for example, in US patent No. 4689338; 4929624; 5266575; 5268376; 5346905; 5352784; 5389640; 5446153; 5482936; 5756747; 6110929; 6194425; 6331539; 6376669; 6451810; 6525064; 6541485; 6545016; 6545017; 6573273; 6656938; 6660735; 6660747; 6664260; 6664264; 6664265; 6667312; 6670372; 6677347; 6677348; 6677349; 6683088; 6756382; 6797718; 6818650 and 77091214; US Patent Publications Nos. 2004/0091491, 2004/0176367 and 2006/0100229; and international patent publications Nos. WO 2005/18551, WO 2005/18556, WO 2005/20999, WO 2005/032484, WO 2005/048933, WO 2005/048945, WO 2005/051317, WO 2005/051324 ,WO 2005/066169 , WO 2005/066170, WO 2005/066172, WO 2005/076783, WO 2005/079195, WO 2005/094531, WO 2005/123079, WO 2005/123080, WO 2006/009826, WO 2006/009832, WO 2006/026760 , WO 2006/028451, WO 2006/028545, WO 2006/028962, WO 2006/029115, WO 2006/038923, WO 2006/065280, WO 2006/074003, WO 2006/083440, WO 2006/086449, WO 2006/091394 , WO 2006/086633, WO 2006/086634, WO 2006/091567, WO 2006/091568, WO 2006/091647, WO 2006/093514 and WO 2006/098852.

Additional examples of small molecule TLR agonists include certain purine derivatives (such as those described in US Patent Nos. 6,376,501 and 6,028,076), certain imidazoquinoline amide derivatives (such as those described in US Patent No. 6,069,149), certain imidazopyridine derivatives (such as those described in US Patent No. 6,518,265 ), certain benzimidazole derivatives (such as those described in US Pat. No. 6,387,938), certain 4-aminopyrimidine derivatives fused to a five-membered nitrogen-containing heterocyclic ring (such as the adenine derivatives described in US Pat. Nos. 6,376,501, 6,028,076 and 6,329,381 and WO 02/08905) and certain 3-.beta.-D-ribofuranosylthiazolo[4,5-d]pyrimidine derivatives (such as those described in US Patent Publication No. 2003/0199461) and certain small molecule immunopotentiating compounds such as those described in e.g. US Patent Publication No. 2005/0136065.

Examples of large TLR agonists include oligonucleotide sequences. Some TLR agonist oligonucleotide sequences contain cytosine-guanine (CpG) dinucleotides and are described, for example, in US Patent Nos. 6194388, 6207646, 6239116, 6339068 and 6406705. Some CpG-containing oligonucleotides may include synthetic immunomodulatory structural motifs, such as those described, for example, in US patent Nos. 6426334 and 6476000. Other TLR agonist nucleotide sequences lack CpG sequences and are described, for example, in international patent publication No. WO 00/75304. Other TLR agonist nucleotide sequences include guanosine- and uridine-rich single-stranded RNA (ssRNA), such as those described, for example, in Heil et al, Science, vol. 303, pp. 1526-1529, Mar. 5, 2004.

Other TLR agonists include biological molecules such as aminoalkylglucosaminide phosphates (AGP) and are described, for example, in US Pat. Nos. 6,113,918, 6,303,347, 6,525,028, and 6,649,172.

TLR agonists also include inactivated pathogens or fractions thereof, which can activate many different types of TLR receptors. Examples of pathogen-derived TLR agonists include BCG, Mycobacterium obuense extract, talimogene laherparepvec (T-Vec) (derived from HSV-1), and Pexa-Vec (derived from vaccinia virus).

In some embodiments, the TLR agonist may be an agonist antibody that specifically binds to a TLR.

Below is a brief description of the various TLRs as well as TLR agonists. The listing of TLR agonists below for a particular TLR should not be interpreted as indicating that said TLR agonist necessarily activates only that TLR (e.g., some molecules can activate multiple types of TLRs or even multiple classes of PRRs). For example, some of the molecules listed below as examples of TLR4 agonists may also be TLR5 agonists.

The terms "TLR1" and "Toll-like receptor 1" refer to any form of the TLR1 receptor, as well as variants, isoforms and species homologues that retain at least a portion of the activity of TLR1. Unless otherwise indicated, for example by reference to human TLR1, TLR1 includes the native TLR1 sequence of all mammalian species, such as human, monkey and mouse. One example of human TLR1 is provided under UniProt accession number Q15399.

As used herein, the term "TLR1 agonist" means any molecule that, upon binding to TLR1, (1) stimulates or activates TLR1, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR1, or (3) increases, promotes or induces TLR1 expression. TLR1 agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, bacterial lipoproteins and derivatives thereof that bind TLR1.

Examples of TLR1 agonists that can be used in the treatments, formulations and applications of the present invention include, for example, bacterial lipoproteins and derivatives thereof, such as SPM-105 (derived from autoclaved mycobacteria), OM-174 (lipid A derivative), OmpS1 (porin from Salmonella typhi ), OmpS1 (porin from Salmonella typhi ), OspA (from Borrelia burgdorferi ), MALP-2 (macrophage-activating lipopeptide-2kD mycoplasma), STF (soluble tuberculosis factor), CU-T12-9, diprozyme and lipopeptides derived from cell wall components such as PAM2CSK4, PAM3CSK4 and PAM3Cys.

TLR1 can form a heterodimer with TLR2 and thus many TLR1 agonists are also TLR2 agonists.

The terms "TLR2" and "Toll-like receptor 2" refer to any form of the TLR2 receptor, as well as variants, isoforms and species homologues that retain at least a portion of the activity of TLR2. Unless otherwise indicated, for example by reference to human TLR2, TLR2 includes the native TLR2 sequence of all mammalian species, such as human, monkey and mouse. One example of human TLR2 is listed under UniProt accession number O60603.

The term "TLR2 agonist" as used herein means any molecule that, upon binding of TLR2, (1) stimulates or activates TLR2, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR2, or (3) increases, promotes or induces expression TLR2. TLR2 agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, bacterial lipoproteins and derivatives thereof that bind to TLR2.

Examples of TLR2 agonists that can be used in the treatments, medicinal products and applications of the present invention include, for example, bacterial lipoproteins (eg, diacylated lipoproteins) and derivatives thereof, such as SPM-105 (derived from autoclaved mycobacteria), OM-174 (lipid A derivative), OmpS1 (porin from Salmonella typhi ), OmpS1 (porin from Salmonella typhi ), OspA (from Borrelia burgdorferi ), MALP-2 (macrophage-activating lipopeptide-2kD of mycoplasma), STF (soluble tuberculosis factor), CU -T12-9, diprovozyme, amplifier and lipopeptides derived from cell wall components such as PAM2CSK4, PAM3CSK4 and PAM3Cys.

TLR2 can form a heterodimer with TLR1 or TLR6, and thus many TLR2 agonists are also TLR1 or TLR6 agonists.

The terms "TLR3" and "Toll-like receptor 3" refer to any form of the TLR3 receptor, as well as variants, isoforms and species homologs that retain at least a portion of the activity of TLR3. Unless otherwise indicated, for example by reference to human TLR3, TLR3 includes the native TLR3 sequence of all mammalian species, such as human, monkey and mouse. One example of human TLR3 is provided under UniProt accession number O15455.

As used herein, the term “TLR3 agonist” means any molecule that, upon binding to TLR3, (1) stimulates or activates TLR3, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR3, or (3) increases, promotes or induces TLR3 expression. TLR3 agonists that can be used in any of the treatments, drugs and applications of the present invention include, for example, nucleic acid ligands that bind to TLR3.

Examples of TLR3 agonists that can be used in the treatments, formulations and applications of the present invention include TLR3 ligands such as synthetic dsRNA, polyinosinic polycytidylic acid ["poly(I:C)"] (available, for example, from InvivoGen at high molecular weight (HMW) and low molecular weight (LMW) drugs), polyadenyl-polyuridylic acid ["poly(A:U)"] (available, for example, from InvivoGen), poly-ICLC (see Levy et al., Journal of Infectious Diseases , vol. 132, no. 4, pp. 434-439, 1975), ampligen (see Jasani et al., Vaccine, vol. 27, no. 25-26, pp. 3401-3404, 2009), hiltonol, rintatolimod and RGC100 (see Naumann et al., Clinical and Developmental Immunology, vol. 2013, article ID 283649).

The terms "TLR4" and "Toll-like receptor 4" refer to any form of the TLR4 receptor, as well as variants, isoforms and species homologs that retain at least a portion of the activity of TLR4. Unless otherwise indicated, for example by reference to human TLR4, TLR4 includes the native sequence of TLR4 from all mammalian species, for example, human, monkey and mouse. One example of human TLR4 is listed under UniProt accession number O00206.

As used herein, the term "TLR4 agonist" means any molecule that, upon binding to TLR4, (1) stimulates or activates TLR4, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR4, or (3) increases, promotes or induces TLR4 expression. TLR4 agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, bacterial lipopolysaccharides (LPS) and derivatives thereof that bind TLR4.

Examples of TLR4 agonists that can be used in the treatments, formulations and applications of the present invention include, for example, bacterial lipopolysaccharides (LPS) and derivatives thereof, such as B:0111 (Sigma), monophosphoryl lipid A (MPLA), 3DMPL (3-O-deacylated MPL), GLA-AQ, G100, AS15, ASO2, GSK1572932A (GlaxoSmithKline, UK).

The terms "TLR5" and "Toll-like receptor 5" refer to any form of the TLR5 receptor, as well as variants, isoforms and species homologs that retain at least a portion of the activity of TLR5. Unless otherwise indicated, for example by reference to human TLR5, TLR5 includes the native sequence of TLR5 from all mammalian species, such as human, monkey and mouse. One example of human TLR5 is listed under UniProt accession number O60602.

As used herein, the term “TLR5 agonist” means any molecule that, upon binding to TLR5, (1) stimulates or activates TLR5, (2) increases, promotes, induces, or prolongs the activity, function, or presence of TLR5, or (3) increases, promotes, or induces TLR5 expression. TLR5 agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, bacterial flagellins and derivatives thereof that bind TLR5.

Examples of TLR5 agonists that can be used in the treatments, medicinal products and uses of the present invention include, for example, bacterial flagellin purified from B. subtilis , flagellin purified from P. aeruginosa , flagellin purified from S. typhimurium , and recombinant flagellin (all of which are available from InvivoGen), entolimod (CBLB502; a pharmacologically optimized flagellin derivative).

The terms "TLR6" and "Toll-like receptor 6" refer to any form of the TLR6 receptor, as well as variants, isoforms and species homologues that retain at least a portion of the activity of TLR6. Unless otherwise indicated, for example by reference to human TLR6, TLR6 includes the native sequence of TLR6 from all mammalian species, for example, human, monkey and mouse. One example of human TLR6 is listed under UniProt accession number Q9Y2C9.

As used herein, the term "TLR6 agonist" means any molecule that, upon binding to TLR6, (1) stimulates or activates TLR6, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR6, or (3) increases, promotes or induces TLR6 expression. TLR6 agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, bacterial lipopeptides and derivatives thereof that bind TLR6.

Examples of TLR6 agonists that can be used in the treatments, formulations and applications of the present invention include, for example, many of the TLR2 agonists presented above, because TLR2 and TLR6 can form a heterodimer. TLR6 can also form a heterodimer with TLR4, and TLR6 agonists can include the various TLR4 agonists presented above.

The terms "TLR7" and "Toll-like receptor 7" refer to any form of the TLR7 receptor, as well as variants, isoforms and species homologs that retain at least a portion of the activity of TLR7. Unless otherwise indicated, for example by reference to human TLR7, TLR7 includes the native sequence of TLR7 from all mammalian species, such as human, monkey and mouse. One example of human TLR7 is listed under UniProt accession number Q9NYK1.

As used herein, the term "TLR7 agonist" means any molecule that, upon binding to TLR7, (1) stimulates or activates TLR7, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR7, or (3) increases, promotes or induces TLR7 expression. TLR7 agonists that can be used in any of the treatments, drugs and applications of the present invention include, for example, nucleic acid ligands that bind to TLR7.

Examples of TLR7 agonists that can be used in the treatments, medicinal products and applications of the present invention include recombinant single-stranded ("ss")RNA, imidazoquinoline compounds such as imiquimod (R837), gardiquimod and resiquimod (R848); loxoribine (7-allyl-7,8-dihydro-8-oxo-guanosine) and related compounds; 7-thia-8-oxoguanosine, 7-deazaguanosine and related guanosine analogues; ANA975 (Anadys Pharmaceuticals) and related compounds; SM-360320 (Sumimoto); 3M-01, 3M-03, 3M-852 and 3M-S-34240 (3M Pharmaceuticals); GSK2245035 (GlaxoSmithKline; 8-oxoadenine molecule), AZD8848 (AstraZeneca; 8-oxoadenine molecule), MEDI9197 (Medimmune; formerly known as 3M-052), ssRNA40 and adenosine analogues such as UC-1V150 (Jin et al., Bioorganic Medicinal Chem Lett (2006) 16:4559–4563, compound 4). Many TLR7 agonists are also TLR8 agonists.

The terms "TLR8" and "Toll-like receptor 8" refer to any form of the TLR8 receptor, as well as variants, isoforms and species homologues that retain at least a portion of the activity of TLR8. Unless otherwise indicated, for example by reference to human TLR8, TLR8 includes the native sequence of TLR8 from all mammalian species, such as human, monkey and mouse. One example of human TLR8 is listed under UniProt accession number Q9NR97.

As used herein, the term “TLR8 agonist” means any molecule that, upon binding to TLR8, (1) stimulates or activates TLR8, (2) increases, promotes, induces, or prolongs the activity, function, or presence of TLR8, or (3) increases, promotes, or induces TLR8 expression. TLR8 agonists that can be used in any of the treatments, drugs and applications of the present invention include, for example, nucleic acid ligands that bind to TLR8.

Examples of TLR8 agonists that can be used in the treatments, medicinal products and applications of the present invention include recombinant single-stranded ssRNA, imiquimod (R837), gardiquimod, resiquimod (R848), 3M-01, 3M-03, 3M-852 and 3M- S-34240 (3M Pharmaceuticals); GSK2245035 (GlaxoSmithKline; 8-oxoadenine molecule), AZD8848 (AstraZeneca; 8-oxoadenine molecule), MEDI9197 (Medimmune; formerly known as 3M-052), poly-G10, motolimod, and the various TLR7 agonists presented above (as described above, many TLR7 agonists are also TLR8 agonists).

The terms "TLR9" and "Toll-like receptor 9" refer to any form of the TLR9 receptor, as well as variants, isoforms and species homologs that retain at least a portion of the activity of TLR9. Unless otherwise indicated, for example with reference to human TLR9, TLR9 includes the native TLR9 sequence of all mammalian species, such as human, monkey and mouse. One example of human TLR9 is listed under UniProt accession number Q9NR96.

As used herein, the term "TLR9 agonist" means any molecule that, upon binding to TLR9, (1) stimulates or activates TLR9, (2) increases, promotes, induces or prolongs the activity, function or presence of TLR9, or (3) increases, promotes or induces TLR9 expression. TLR9 agonists that can be used in any of the treatments, medicinal products and applications of the present invention include, for example, nucleic acid ligands that bind to TLR9.

Examples of TLR9 agonists that can be used in the treatments, medicinal products and applications of the present invention include unmethylated CpG-containing DNA, immunostimulatory oligodeoxynucleotides (ODNs) such as CpG-containing ODNs such as CpG24555, CpG10103, CpG7909 (PF-3512676 /agatolimod), CpG1018, AZD1419, ODN2216, MGN1703, SD-101, 1018ISS and CMP-001. TLR9 agonists also include nucleotide sequences containing synthetic cytosine phosphate 2'-deoxy-7-deazaguanosine dinucleotide (CpR) (Hybridon, Inc.), dSLIM-30L1, and immunoglobulin-DNA complexes. Examples of TLR9 agonists are described in WO2003/015711, WO2004/016805, WO2009/022215, PCT/US95/01570, PCT/US97/19791 and US Patent Nos. 8552165, 6194388 and 6239116, each of which is hereby incorporated herein. as a reference for all purposes.

RLRs include various cytosolic PRRs that sense, for example, dsRNA. Examples of RLRs include, for example, retinoic acid-inducible gene I (RIG-I), melanoma differentiation-associated gene 5 (MDA-5), and Laboratory of Genetics and Physiology protein 2 (LGP2).

As used herein, the term "RLR agonist" means any molecule that, upon binding to RLR, (1) stimulates or activates RLR, (2) increases, promotes, induces, or prolongs the activity, function, or presence of RLR, or (3) increases, promotes, or induces RLR expression. RLR agonists that can be used in any of the treatments, medicinal products and applications of the present invention include, for example, nucleic acids and derivatives thereof that bind to RLR, and agonistic monoclonal antibodies (mAbs) that specifically bind to RLR.

Examples of RLR agonists that can be used in the treatments, formulations and applications of the present invention include, for example, short double-stranded RNA with uncapped 5'-triphosphate (RIG-I agonist); poly I:C (MDA-5 agonist) and BO-112 (MDA-A agonist).

NLRs include various PRRs that define, for example, damage-associated molecular pattern molecules (DAMPs). NLRs include the NLRA-A, NLRB-B, NLRC-C, and NLRP-P subfamilies. Examples of NLRs include, for example, NOD1, NOD2, NAIP, NLRC4 and NLRP3.

As used herein, the term "NLR agonist" means any molecule that, upon binding to an NLR, (1) stimulates or activates an NLR, (2) increases, promotes, induces, or prolongs the activity, function, or presence of an NLR, or (3) increases, promotes, or induces NLR expression. NLR agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, DAMPs and derivatives thereof that bind to NLR, and agonistic monoclonal antibodies (mAbs) that specifically bind to NLR.

Examples of NLR agonists that can be used in the treatments, formulations and applications of the present invention include, for example, liposomal muramyl tripeptide/mifamurtide (NOD2 agonist).

CLRs include various PRRs that sense, for example, carbohydrates and glycoproteins. CLRs include transmembrane CLRs and secreted CLRs. Examples of CLRs include, for example, DEC-205/CD205, macrophage mannose receptor (MMR), Dectin-1, Dectin-2, Mincle, DC-SIGN, DNGR-1 and mannose binding lectin (MBL).

As used herein, the term "CLR agonist" means any molecule that, upon binding to CLR, (1) stimulates or activates CLR, (2) increases, promotes, induces, or prolongs the activity, function, or presence of CLR, or (3) increases, promotes, or induces CLR expression. CLR agonists that can be used in any of the treatments, formulations and applications of the present invention include, for example, carbohydrates and derivatives thereof that bind to CLR and agonistic monoclonal antibodies (mAbs) that specifically bind to CLR.

Examples of CLR agonists that can be used in the treatments, formulations and applications of the present invention include, for example, MD-fraction (purified soluble beta-glucan extract from Grifola frondosa ) and Imprime PGG (beta 1,3/1,6- glucan PAMP derived from yeast).

The STING protein functions as a cytosolic DNA sensor and adapter protein in type 1 interferon signaling. The terms "STING" and "interferon gene stimulator" refer to any form of the STING protein, as well as variants, isoforms and species homologs that retain at least a portion of STING activity . Unless otherwise indicated, for example by reference to human STING, STING includes the native STING sequence of all mammalian species, such as human, monkey and mouse. One example of human TLR9 is listed under UniProt accession number Q86WV6. STING is also known as TMEM173.

As used herein, "STING" means any molecule that, upon binding to TLR9, (1) stimulates or activates STING, (2) increases, promotes, induces or prolongs the activity, function or presence of STING, or (3) increases, promotes or induces expression STING. STING agonists that can be used in any of the treatments, medicinal products and applications of the present invention include, for example, nucleic acid ligands that bind to STING.

Examples of STING agonists that can be used in the treatments, medicinal products and applications of the present invention include various immunostimulatory nucleic acids such as synthetic double-stranded DNA, cyclic di-GMP, cyclic GMP-AMP (cGAMP), synthetic cyclic dinucleotides (CDN) , such as MK-1454 and ADU-S100 (MIW815), and small molecule compounds such as P0-424.

Other PRRs include, for example, DNA-dependent activator of IFN-regulatory factors (DAI) and Absent in Melanoma 2 (AIM2) protein.

Immunostimulatory cytokines include various signaling proteins that stimulate the immune response, such as interferons, interleukins, and hematopoietic growth factors.

Examples of immunostimulatory cytokines include GM-CSF, G-CSF, IFN-alpha, IFN-gamma; IL-2 (eg, denileukin diftitox), IL-6, IL-7, IL-11, IL-12, IL-15, IL-18, IL-21 and TNF-alpha.

The immunostimulatory cytokines may be in any suitable format. In some embodiments, the immunostimulatory cytokine may be a recombinant version of the wild-type cytokine. In some embodiments, the immunostimulatory cytokine may be a mutein having one or more amino acid changes compared to the corresponding wild-type cytokine. In some embodiments, the immunostimulatory cytokine can be included in a chimeric protein containing the cytokine and at least one other functional protein (eg, an antibody). In some embodiments, the immunostimulatory cytokine may be covalently linked to a drug/agent (eg, any drug/agent described elsewhere herein as a possible component of an ADC).

Tumor vaccines include various compositions that contain tumor-associated antigens (or that can be used to produce a tumor-associated antigen in an individual), and thus can be used to provoke an immune response in an individual that will be directed to tumor cells containing the tumor-associated antigen.

Examples of materials that can be included in a tumor vaccine include attenuated cancer cells, tumor antigens, antigen presenting cells such as dendritic cells to which tumor antigen has been added or nucleic acids encoding tumor-associated antigens. In some embodiments, a tumor vaccine can be produced using a patient's own cancer cells. In some embodiments, a tumor vaccine can be produced using biological material other than the patient's own cancer cells.

Tumor vaccines include, for example, sipuleucel-T and talimogene laherparepvec (T-VEC).

Immune cell therapy involves treating a patient with immune cells capable of targeting malignant cells. Immune cell therapies include, for example, tumor-infiltrating lymphocytes (TILs) and chimeric antigen receptor T cells (CAR-T cells).

Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkylsulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa and uredopa; ethyleneimines and methylmelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; acetogenins (in particular bullatacin and bullatacinone); camptothecin (including a synthetic analogue of topotecan); bryostatin; kallistatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin and bizelesin); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including synthetic analogues, KW-2189 and CBI-TMI); eleutherobin; pancratistatin; sarcodictin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphasine, holophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembiquin, phenesterine, prednimustine, trofosfamide, uramustine; nitrosoureas such as carmustine, chlorosotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as enediyne antibiotics (e.g. calicheamycin, in particular calicheamycin gamma 1I and calicheamycin phi I1, see, for example, Agnew, Chem. Intl. Ed. Engl., 33:183-186 (1994); dinemycin, including dinemycin A; bisphosphonates such as clodronate; esperamycin; and neocarcinostatin chromophore and related chromoprotein chromophores of enediyne antibiotics), aclacinomysins, actinomycin, outramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carcinophilin, chromomycins, dactinomycin, daunorubicin , detorubicin, 6 -diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), pegylated liposomal doxorubicin, epirubicin, ezorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprin, thioguanine; pyrimidine analogues such as ancytabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocytabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepithiostane, testolactone; anti-adrenal drugs such as aminoglutemide, mitotane, trilostane; a folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatrexate; defofamine; demecolcine; diaziquon; eflornithine; elliptinium acetate; epothilone; ethoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamole; nitracrine; pentostatin; phenomet; pirarubicin; losoxantrone; podophyllic acid; 2-ethylhydrazide; procarbazine; razoxane; rhizoxin; sisofuran; spirogermanium; tenuazonic acid; triaziquon; 2,2′,2″-trichlorotriethylamine; trichothecenes (particularly T-2 toxin, verracurine A, roridin A and anguidine); urethane; vindesine; dacarbazine; mannomustin; mitobronitol; mitolactol; pipobromance; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, for example, paclitaxel and doxetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; Novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above compounds. Also included are antihormonal agents that regulate or inhibit the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifene, trioxifene, keousifene, LY117018, onapristone, and toremifene (Fareston ); aromatase inhibitors, which inhibit the aromatase enzyme that regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole, vorozole, letrozole and anastrozole; and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; KRAS inhibitors; MCT4 inhibitors; MAT2a inhibitors; tyrosine kinase inhibitors such as sunitinib, axitinib; alk/c-Met/ROS inhibitors such as crizotinib, lorlatinib; mTOR inhibitors such as temsirolimus, gedatolisib; src/abl inhibitors such as bosutinib; cyclin-dependent kinase (CDK) inhibitors such as palbociclib, PF-06873600; erb inhibitors such as dacomitinib; PARP inhibitors such as talazoparib; SMO inhibitors such as glasdegib, PF-5274857; EGFR T790M inhibitors such as PF-06747775; EZH2 inhibitors such as PF-06821497; PRMT5 inhibitors such as PF-06939999; TGFRβr1 inhibitors such as PF-06952229; and pharmaceutically acceptable salts, acids or derivatives of any of the above compounds. In specific embodiments, such additional therapeutic agent is bevacizumab, cetuximab, sirolimus, panitumumab, 5-fluorouracil (5-FU), capecitabine, tivozanib, irinotecan, oxaliplatin, cisplatin,trifluridine, tipiracil, leucovorin, gemcitabine, regorafinib, or erlotinib hydrochloride.

In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination with one or more other therapeutic agents targeting an immune checkpoint modulator or costimulatory agent, such as, but not limited to, a CTLA-targeting agent -4, LAG-3, B7-H3, B7-H4, B7-DC (PD-L2), B7-H5, B7-H6, B7-H8, B7-H2, B7-1, B7-2, ICOS, ICOS-L, TIGIT, CD2, CD47, CD80, CD86, CD48, CD58, CD226, CD155, CD112, LAIR1, 2B4, BTLA, CD160, TIM1, TIM-3, TIM4, VISTA (PD-H1), OX40, OX40L , GITR, GITRL, CD70, CD27, 4-1BB, 4-BBL, DR3, TL1A, CD40, CD40L, CD30, CD30L, LIGHT, HVEM, SLAM (SLAMF1, CD150), SLAMF2 (CD48), SLAMF3 (CD229), SLAMF4 (2B4, CD244), SLAMF5 (CD84), SLAMF6 (NTB-A), SLAMCF7 (CS1), SLAMF8 (BLAME), SLAMF9 (CD2F), CD28, CEACAM1 (CD66a), CEACAM3, CEACAM4, CEACAM5, CEACAM6, CEACAM7 , CEACAM8, CEACAM1-3AS, CEACAM3C2, CEACAM1-15, PSG1-11, CEACAM1-4C1, CEACAM1-4S, CEACAM1-4L, IDO, TDO, CCR2, CD39-CD73-adenosine (A2AR) pathway, BTK, TIK, CXCR2 , CXCR4, CCR4, CCR8, CCR5, CSF-1 or a modulator of the innate immune response.

In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination, for example, with an anti-CTLA-4 antagonist antibody, such as, for example, ipilimumab; an anti-LAG-3 antagonist antibody such as BMS-986016 and IMP701; antagonistic antibody against TIM-3; an anti-B7-H3 antagonist antibody such as, for example, MGA271; antagonistic antibody against VISTA; antagonistic antibody against TIGIT; antibody; anti-CD80 antibody; anti-CD86 antibody; antagonistic antibody against B7-H4; agonistic antibody against ICOS; agonistic antibody against CD28; a modulator of the innate immune response (eg, TLR, KIR, NKG2A) and an IDO inhibitor.

In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination with an OX40 agonist, such as, for example, an anti-OX-40 agonist antibody. In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination with a GITR agonist, such as, for example, an anti-GITR agonist antibody such as, but not limited to, TRX518. In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination with an IDO inhibitor. In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination with a cytokine therapy such as, but not limited to, IL-15, CSF-1, M-CSF-1, etc.

In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody is used in combination with one or more other therapeutic antibodies, such as, but not limited to, an anti-CD19, CD22, CD40, CD52, or CCR4 antibody.

In some embodiments, the anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody composition contains at least one additional agent, such as bevacizumab, cetuximab, sirolimus, panitumumab, 5-fluorouracil (5-FU), capecitabine, tivozanib, irinotecan, oxaliplatin, cisplatin, trifluridine, tipiracil, leucovorin, gemcitabine and erlotinib hydrochloride.

In some embodiments, an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody may be co-administered or administered sequentially before or after another agent at intervals ranging from minutes to weeks. In embodiments, if other agents and/or proteins or polynucleotides are administered separately, it will generally be ensured that no significant period of time has passed between each administration, such that the agent and composition of the present invention will still be able to cause predominantly combined effect on an individual. In such cases, it is envisaged that both agents can be administered within approximately 12-24 hours of each other, and more preferably within approximately 6-12 hours of each other. However, in some cases, it may be desirable to significantly increase the period of time for administration, with several days (2, 3, 4, 5, 6, or 7) to several weeks (1, 2, 3, 4, 5, 6) between appropriate administrations. , 7 or 8).

In some embodiments, the anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody composition is combined with a treatment regimen further comprising a conventional therapy selected from the group consisting of: surgery, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, angiogenesis inhibition and palliative care.

The composition used in the present invention may further contain pharmaceutically acceptable carriers, excipients or stabilizers (Remington: The Science and practice of Pharmacy 21st Ed., 2005, Lippincott Williams and Wilkins, Ed. K. E. Hoover) in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients or stabilizers are non-toxic to recipients in dosages and concentrations and may contain buffers such as phosphate, citrate and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkylparabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrans; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; complex compounds with metals (for example, Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONIC™ or polyethylene glycol (PEG). Pharmaceutically acceptable excipients are further provided herein.

In one aspect, the present invention provides a method for producing an anti-GUCY2c antibody or a bispecific antibody as provided herein, comprising culturing a host cell under conditions leading to the production of an anti-GUCY2c antibody or a bispecific antibody as provided herein, and purifying the antibody or bispecific antibody from culture.

In another aspect, the present invention relates to the use of an anti-GUCY2c antibody, bispecific antibody, pharmaceutical composition, polynucleotide, supernatant, vector or host cell provided herein in the manufacture of a medicament for the detection, diagnosis and/or treatment of a GUCY2c-associated disorder.

An additional aspect of the invention relates to a kit containing an anti-GUCY2c antibody or an anti-CD3-GUCY2c bispecific antibody as described herein above and instructions for use in accordance with any of the methods of the invention provided herein. Typically, these instructions include a description of the administration of an anti-GUCY2c antibody or a bispecific anti-CD3-GUCY2c antibody for the therapeutic treatment described above. This kit contains any pharmaceutical composition presented in the present description. The pharmaceutical compositions and other reagents may be present in the kits in any convenient form, such as, for example, a solution or powder.

In another aspect, the kit further contains one or more other prophylactic or therapeutic agents that can be used in the treatment of cancer in one or more containers. In one embodiment, the other prophylactic or therapeutic agent is a chemotherapeutic agent. In other aspects, the prophylactic or therapeutic agent is a biological or hormonal therapeutic agent.

Certain aspects of the pharmaceutical compositions, prophylactic or therapeutic agents of the invention are preferably tested in vitro , in a cell culture system and in an animal model organism, such as a rodent model system, for the desired therapeutic activity before use in humans.

The toxicity and efficacy of the prophylactic and/or therapeutic methods of the present invention can be determined by standard pharmaceutical methods in cell culture or experimental animals, for example, to determine the LD ₅₀ (dose lethal in 50% of the population) and ED ₅₀ (dose therapeutically effective in 50% of the population). % of the population). The dose relationship between toxic and therapeutic effects represents the therapeutic index and can be expressed as the ratio LD ₅₀ /ED ₅₀ . Prophylactic and/or therapeutic agents demonstrating large therapeutic indices are preferred.

In addition, any assay known to those skilled in the art can be used to evaluate the prophylactic and/or therapeutic utility of the therapeutic agents or combination therapy provided herein for the treatment or prevention of cancer.

Instructions regarding the use of the anti-GUCY2c antibody or anti-CD3-GUCY2c bispecific antibody provided herein generally include information regarding the dose, dosage regimen, and route of administration for the intended treatment. Containers may be single dose, multi-dose packages or sub-single dose. The instructions provided in the kits of the invention are typically written instructions on a label or package insert (eg, a piece of paper included in the kit), but machine-readable instructions (eg, instructions on a magnetic or optical storage device) are also acceptable.

The kits of the present invention are provided in suitable packaging. Suitable packaging includes, but is not limited to, vials, ampoules, tubes, carboys, jars, flexible packaging (eg, sealed mylar or plastic bags), and the like. for each pharmaceutical composition and other included reagents, such as buffers, balanced salt solutions, etc., for use in administering the pharmaceutical compositions to individuals. Packages are also provided for use in combination with a specific device, such as an inhaler, a nasal delivery device (eg, an atomizer), or an infusion device such as a minipump. The kit may have a sterile entry port (eg, the container may be an intravenous solution bag or a vial having a stopper pierced by a hypodermic needle). The container may also have a sterile entry port (eg, the container may be an intravenous solution bag or a vial having a stopper pierced by a hypodermic needle). At least one active agent in the composition is an anti-GUCY2c antibody or a bispecific anti-CD3-GUCY2c antibody. The container may further contain a second pharmaceutically active agent.

Typically, the kit contains a container and a label or package inserts on or attached to the container.

Deposition of biological material

Representative materials of the present invention are deposited in the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209, USA, as of February 13, 2018. Chain Vector A of GUCY2C-1608 (VH-cavity SEQ ID NO: 220), having ATCC accession number PTA-124943, contains a DNA insert (SEQ ID NO: 247) encoding the VH-cavity chain of the bispecific antibody GUCY2C-1608, and the chain vector B GUCY2C-1608 (VL overhang SEQ ID NO: 216), having ATCC accession number PTA-124944, contains a DNA insert (SEQ ID NO: 246) encoding the VL overhang strand of the bispecific antibody GUCY2C-1608.

The deposit was carried out in accordance with the provisions of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure and Regulations Thereunder (Budapest Treaty). This ensures that a viable deposit culture is maintained for a period of 30 years from the date of deposit. The deposit will be made available to ATCC under the Budapest Treaty and is subject to an agreement between Pfizer Inc. and an ATCC providing for the continued and unrestricted availability of the progeny of the deposit culture to the public upon the issuance of a permanent U.S. patent or the issuance of any U.S. patent application or foreign patent application, whichever occurs first, and making the progeny available to anyone designated by the Patent and Trademark Office US marks as entitled to do so under 35 U.S.C. Section 122. and Agency rules consistent therewith (including 37 C.F.R. section 1.14 with specific reference to 886 OG 638).

The transferee of this application agrees that if the deposited materials perish, are lost, or are destroyed during cultivation under suitable conditions, the materials will be promptly replaced upon notice with the same materials. The availability of the material deposited should not be construed as a license to practice the present invention in violation of the rights granted by any government authority under patent law.

The following examples of specific aspects of the present invention are presented for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

Examples

Example 1: Immunization and hybridoma production of mouse anti-GUCY2c antibodies

Recombinant human GUCY2c immunogen protein or cells overexpressing human GUCY2c on their surface (SEQ ID NO: 224) was injected into Balb/c mice to produce hybridomas. Eight-week-old female Balb/c mice were immunized with huGUCY2c-mIgG2a soluble protein or the 300.19 cell line expressing human GUCY2c (SEQ ID 224/225). For mice immunized with the recombinant huGUCY2c protein, the animals were dosed according to the RIMMS protocol (Kilpatrick, et al. Hybridoma. 1997. 16:381-389). Briefly, mice were immunized six times with recombinant human GUCY2c-mouse IgG2a Fc fusion protein (SEQ ID NO: 230) over two weeks by subcutaneous injection. The adjuvants used were Freund's complete and incomplete adjuvants. For cell-based immunization, Balb/c mice were immunized with 5 x 10 ⁶ 300.19 cells overexpressing human GUCY2c twice weekly for one month without adjuvant via ip injection. To determine mouse anti-GUCY2c antibody titers, blood samples from immunized animals were obtained and GUCY2c-specific titers were examined by enzyme-linked immunosorbent assay (ELISA) using recombinant human GUCY2c mIgG2a-Fc (SEQ ID NO: 230) and cynomolgus GUCY2c-mIgG2a-Fc (SEQ ID NO: 234). Animals with the highest titers of GUCY2c were selected for hybridoma production. In the case of mice immunized with the soluble recombinant protein, LN lymphocytes and splenocytes were used for fusion, while in the case of mice immunized with 300.19/hGUCY2c, only splenocytes were used for fusion. To obtain a hybridoma, splenocytes and/or lymphocytes from lymph nodes were fused with P3X myeloma in a 1:1 ratio using PEG. The fusion cells were selected in the presence of HAT for seven days, after which the hybridomas were maintained in HT-containing media prior to screening. To identify anti-huGUCY2c antibody-specific hybridomas, hybridoma supernatants were screened for anti-huGUCY2c IgG reactivity by ELISA. Potential anti-GUCY2c hybridomas were then pooled and assayed for reactivity against human and cynomolgus GUCY2c on the surface of transfected 300.19 cell lines and primary tumor cell lines expressing GUCY2c. Briefly, 300.19 cell lines expressing various GUCY2c proteins (SEQ ID NOs: 224, 226 and 228) were seeded at 4 x 10 ⁴ cells/well in sterile 96-well tissue culture plates (Becton Dickinson) on day 1 and incubated at 37°C/5% CO ₂ for 1-2 days until a confluent monolayer was achieved. Cells were washed 4 times with PBS+Ca ²⁺ and Mg ²⁺ and blocked for 1 hour at room temperature with 3% milk/1% BSA/PBS+Ca ²⁺ and Mg ²⁺ . Serially diluted samples (1:3 in blocking buffer) were applied to the plate. The plates were incubated at room temperature for 2 hours before washing with PBS+Ca ²⁺ and Mg ²⁺ . HRP-conjugated secondary antibody, goat anti-mouse Fc IgG (Thermo Scientific), diluted (1:4,000) in blocking buffer, was then added and incubated with the cells for 1 hour. The plates were washed with PBS+Ca ²⁺ and Mg ²⁺ before development with TMB substrate solution for 10 minutes, and then 0.18M⋅H ₂ SO ₄ was added to stop the reaction. The absorbance was measured at OD 450 nM, the data was plotted and analyzed using Microsoft Excel and Graphpad-Prism software. Hybridomas that specifically bind to human and cynomolgus GUCY2c were identified.

Example 2: Binding of Anti-GUCY2c Hybridoma Antibodies in Flow Cytometry and Immunohistochemistry Assays

Anti-GUCY2c hybridoma antibodies were screened by flow cytometry for cell surface binding to human GUCY2c on T84 and HT55 tumor cells. HT29 cells were used as negative controls. Cells were dissociated with cell dissociation buffer (Sigma) and blocked on ice with PBS+3% BSA. Cells were stained on ice with primary antibody for 1 hour and then washed with cold PBS. 10 μg/ml secondary antibody (PE-conjugated anti-mouse secondary antibody, Jackson Immunoresearch) was added on ice for an hour. Cells were washed again with cold PBS, resuspended in PBS, stained with DAPI to distinguish live/dead cells, and assayed for GUCY2c binding using a BD LSRII Fortessa. Several clones were identified that showed specific binding to GUCY2c-expressing cell lines (T84 and HT55) and lack of binding to GUCY2c-negative cells (HT29).

Hybridomas from BALB/c mice immunized against GUCY2c were tested for immunoreactivity in formalin-fixed paraffin-embedded cell pellets. Cell pellets obtained from this screen consisted of parental 300.19 cells, 300.19 cells overexpressing mouse, cynomolgus or human GUCY2c, one human colorectal cancer cell line expressing high endogenous levels of GUCY2c (T84), and one human colorectal cancer cell line negative for GUCY2c expression (HT29). Cell lines were fixed for 24 h in 10% neutral buffered formalin (Thermo Scientific) and centrifuged at 300 × g for 4 minutes (min) to pellet. Formalin was removed and cell pellets were carefully resuspended with prewarmed 50ºC Histogel (Thermo Scientific). Cell pellets embedded in Histogel were cooled at 4°C for 1-2 hours before processing overnight in a VIP automated histoprocessor (Tissue-Tek). The treated cell pellets were embedded in paraffin. Five-μm sections of cell micropanels were cut, transferred to a water bath, and mounted on Superfrost Excell slides (Fisher). The glasses were allowed to dry overnight. After deparaffinization and rehydration of tissue sections, thermally induced epitope unmasking was performed using a Retriever 2100 digester autoclave (Electron Microscopy Sciences) in Borg Decloaker buffer, pH 9.5 (Biocare Medical), or citrate buffer, pH 6.0 (Thermo Scientific), followed by cooling to room temperature (RT). Endogenous peroxidase activity was inactivated using Peroxidazed 1 (Biocare Medical) for 10 min. Nonspecific protein interactions were blocked for 10 min using Background Punisher (Biocare Medical). Each hybridoma was incubated without dilution for 1 hour under conditions of thermally induced epitope unmasking. Sections were washed in TBS and hybridoma binding was determined using Envision+ Mouse HRP (DAKO) for 30 min. Slides were washed in TBS and immunoreactivity was determined using the Betazoid DAB Chromogen Kit (Biocare Medical) for 5 min, followed by rinsing with distilled water. Immunostained media were quickly counterstained with hematoxylin CAT (Biocare Medical), rinsed with tap water, dehydrated with a battery of alcohols, cleared in xylene, and coverslipped with Permount histology media (FisherChemicals). The slides were evaluated by a veterinary pathologist for immunoreactivity analysis. This screen identified clones with immunoreactivity for human, cynomolgus and mouse GUCY2c.

Results regarding the binding of anti-GUCY2c hybridoma antibodies by flow cytometry and immunohistochemistry (IHC) are shown in Table 11.

Table 11

Clone T84+ HT55+ HT29- binding by flow cytometry IHC binding (human GUCY2c) IHC binding (GUCY2c cynomolgus) IHC binding (mouse GUCY2c) 1A7 + + + - 2D4 + 4F2 + 4F12 + 4G3 + 5G7 + 9H3 + + + + 20D11 + 3H6 + 18E10 +

Example 3: Quantitative Analysis of GUCY2c Expression Levels in Cell Lines

GUCY2c receptor density on cell lines was measured using clone GUCY2C-9H3 conjugated 1:1 to phycoerythrin (Thermo Scientific). 1x10 ⁵ cells were stained with 10 μg/ml PE-labeled anti-GUCY2c mAb for saturable binding. Cells were washed and resuspended in FACS buffer with DAPI and analyzed using a BD LSRII Fortessa with FACS Diva software. QuantiBRITE PE-labeled beads (BD) were used under the same PE loading conditions to calculate the number of PE-labeled antibodies per cell (ABC) based on the geometric mean of PE fluorescence intensity. Measurements of GUCY2c receptor densities in human tumor cell lines are shown in Table 12. Cell lines with GUCY2c receptor densities were characterized in this assay and used in subsequent assays (eg, cytotoxicity assays) to evaluate the functional activity of anti-CD3-GUCY2c bispecific antibodies.

Table 12

Cell line Antibody binding capacity (ABC)/cell T84 8066.6 LS1034 6007.7 SNU16 3029.3 HT55 2811.5 SW1116 2650.7 CACO-2 4214.6 JHH7 2290 LS174T 1666.8 ASPC-1 495.6

Example 4: Cloning and Sequencing of Anti-GUCY2c Hybridoma Antibodies

Hybridoma supernatants with confirmed cell surface binding activity were subcloned into mouse IgG1 format. RNA was isolated and variable (V) region DNA sequences were obtained from the expressed antibodies by reverse transcription-polymerase chain reaction (RT-PCR) cloning as described below.

One to five million subcloned hybridoma cells were homogenized for total RNA isolation using the QIAGEN RNAeasy Mini kit. First-strand cDNA was then prepared using the SuperScript III RT kit (Invitrogen). Double-stranded cDNA for the anti-GUCY2c IgG variable regions was then prepared and amplified by PCR using primers for the mouse IgG heavy chain (IgG1, IgG2a, IgG2b) and light chain (kappa or lambda) constant regions as described below. The PCR cycle conditions were as follows: 1 cycle at 95°C for 1 min; 25 cycles of 95°C for 1 min, 63°C for 1 min, and 72°C for 1 min. The resulting RT-PCR products were cloned into the TOPO-Blunt cloning vector (Invitrogen) and sequenced.

Residues in the variable domain were numbered according to Kabat, which is the numbering system used for heavy chain variable domains or light chain variable domains of antibody compilation (Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD). Using this numbering system, the exact linear amino acid sequence may contain fewer or more amino acids, corresponding to a truncation or insertion in the FR or CDR of the variable domain. For example, a heavy chain variable domain may include a single amino acid insertion (Kabat residue 52a) after H2 residue 52 and inserted residues (eg, Kabat residues 82a, 82b and 82c) after heavy chain framework residue (FR) 82. Kabat numbering of residues can be determined for a specified antibody by alignment in regions of homology of the antibody sequence with a “standard” Kabat numbered sequence. Various Kabat numbering algorithms are available. Herein, the algorithm embodied in version 2.3.3 of Abysis (www.abysis.org) is used to assign Kabat numbering to the CDR1 VL, CDR2 VL, CDR3 VL, CDR2 VH and CDR3 VH variable regions. The AbM definition is used for CDR1 VH. FR residues are residues of the antibody variable domain other than CDR residues. The VH or VL domain scaffold contains four scaffold subregions, FR1, FR2, FR3 and FR4, interspersed with CDRs in the following order: FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4.

Chimeric T cell bispecific proteins with mouse V regions and modified human IgG constant regions (hIgG1) were prepared as follows: V region cDNA from parental mouse TOPO vectors was subcloned into mammalian expression vectors consisting of a first and second heterodimerization promoting domain, each which contains a cysteine linker (linker 3), such as GCPPCP (SEQ ID NO: 192), and a knob Fc chain (SEQ ID NO: 188) or a trench Fc chain (SEQ ID NO: 189) . In this case, the heavy chain variable region (VH) of the mouse anti-GUCY2c antibody (SEQ ID NOs: 11, 19, 26, 33 and 41) was fused in frame to the light chain variable region (VL) of the anti-human CD3 antibody (SEQ ID NO: 84), separated by a linker GGGSGGGG (SEQ ID NO: 190), and then another linker GCPPCP (SEQ ID NO: 192), connecting to the dimpled Fc chain (SEQ ID NO: 189). The VL regions against mouse GUCY2c (SEQ ID NO: 92, 100, 104, 106, 112 and 119) were fused in frame to the linker GGGSGGGG (SEQ ID NO: 190), the VH region against human CD3 (SEQ ID NO: 1) , followed by a GCPPCP linker (SEQ ID NO: 192) and an overhang Fc chain (SEQ ID NO: 188). Fragments were amplified by standard PCR using primers containing 15-25 nucleotide overhangs with homology to the host vector for isothermal assembly (ITA) cloning. The fragments were recombined into the above linearized vectors using the Infusion ITA cloning kit (Clontech).

The ligation products were then transformed into competent E. coli DH5α (Life Technologies) and grown overnight at 37°C on agarose plates containing 100 μg/ml carbenicillin. Colonies were counted, picked, and grown overnight in YT broth with 100 μg/ml carbenicillin, and DNA was isolated by standard methods. All DNA constructs were then sequenced on both strands prior to expression in mammalian cells using conventional sequencing methods.

Example 5: Expression and Purification of Chimeric Anti-CD3-GUCY2c Bispecific Antibodies

Complementary pair constructs (12.5 μg of each strand) were cotransfected into 25 ml of log-phase culture containing 1 million cells/ml HEK 293 cells using the ExpiFectamine™ 293 transfection kit (Life Technologies). 24 hours after transfection, the transfection enhancer ExpiFectamine was added and cells were allowed to grow for an additional 4–5 days before harvesting. Depleted cultures were then harvested, centrifuged to remove cellular debris, and then passed through a 20-μm filter.

Purified conditioned media containing anti-CD3-GUCY2c bispecific antibodies were then purified using Protein A affinity chromatography. Samples were injected into 0.45 mL microcolumns (Repligen) pre-packed with MabSelect SuRe™ Protein A resin (GE Healthcare) using liquid manipulator (Tecan). Bound protein was washed with PBS, pH 7.2, then eluted with 20 mM citric acid, 150 mM sodium chloride, pH 3.5, and neutralized with 2 M Tris, pH 8.0. Samples were then desalted in PBS, pH 7.2, using G25 Sephadex dropping columns (GE Healthcare) according to the manufacturer's instructions. Purified proteins were analyzed for purity using analytical-scale size exclusion chromatography using a Mab HTP column (Tosoh Bioscience) and an Aglient 1200 HPLC according to the manufacturer's instructions. Concentrations were determined by measuring absorbance at OD280 nm using a microspectrophotometer (Trinean).

Example 6: Recombinant Protein Binding Assay

Proteins were screened for binding to human, cynomolgus and mouse recombinant proteins by ELISA. Purified human, cynomolgus, mouse, or human CD3 epsilon delta ECDs were coated on 384-well Nunc-Maxisorb ELISA plates at 1 μg/ml in 20 μl in PBS-CMF (without calcium and magnesium) at 4°C. The plates were washed 3 times with PBS+0.05% TWEEN20 and blocked with PBS+3% milk for 1 h and shaken at RT. The blocking solution was removed, and samples serially diluted in the blocking solution were added and incubated for 1 h with shaking at RT. The plates were washed 3 times as described above, and a secondary antibody was added (goat anti-human Fc IgG-HRP - Southern Biotech L2047-05, 1:5000; goat anti-mouse Fc IgG-HRP - Thermo Scientific, diluted 1:4000; or mouse anti-penta-HIS-HRP antibody, Qiagen, 1:1000), followed by shaking for 1 hour at RT. The plates were washed as described above and signal developed using TMB, the reaction was stopped with H ₂ SO ₄ and the absorbance was read at 450 nm using an Envision plate reader (Perkin Elmer). An alternative detection method was also periodically used instead of TMB/H ₂ SO ₄ . After addition and initial wash, 20 μl per well of anti-human IgG europium conjugate (Perkin Elmer EU-N1) diluted 1:1000 in DELFIA assay buffer (Perkin Elmer 1244-330) was added and incubated at RT for 1 h. Then The plates were washed 3 times with 1× DELFIA wash solution (Perkin Elmer 1244-114), 20 µl per well of enhancement solution (Perkin Elmer 1244-105) was added and incubated for 30 minutes. Time-resolved fluorescence was then measured at 320 and 615 nm using an EnVision® Plate Reader Spectrophotometer (EnVision® Multichannel Plate Reader Spectrophotometer, Perkin Elmer) according to the manufacturer's instructions.

To test bispecific binding to GUCY2c and CD3, a sandwich ELISA was performed. Human CD3 epsilon delta (SEQ ID NO: 242) was coated on 384-well Nunc-Maxisorb ELISA plates at 1 μg/ml in 20 μl PBS-CMF (without calcium and magnesium) at 4°C. The plates were washed 3 times with PBS+0.05% TWEEN20 and blocked with PBS+3% milk for 1 h with shaking at RT. The blocking solution was removed, samples serially diluted in the blocking solution were added, and incubated for 1 h with shaking at RT. The plates were washed 3 times as described above, and purified human, cynomolgus, or mouse Fc GUCY2c fusion protein was added to the plate at 1 μg/ml and incubated at RT with shaking for 1 h. The plates were washed 3 times. as described above, and secondary antibody (goat anti-mouse Fc IgG-HRP - Thermo Scientific, diluted 1:4000) was added, followed by shaking for 1 h at RT. The plates were washed as described above and signal developed using TMB, the reaction was stopped with H ₂ SO ₄ and the absorbance was read at 450 nm using an EnVision® plate reader (Perkin Elmer). An alternative detection method was also periodically used instead of TMB/H ₂ SO ₄ . After adding and washing the purified human, cynomolgus, or mouse GUCY2c ECD fusion protein-mouse IgG2a Fc fusion protein, 20 μl per well of anti-mouse IgG europium conjugate (Perkin Elmer EU-N1) diluted 1:1000 in DELFIA assay buffer (Perkin Elmer) was added 1244-330) and incubated at RT for 1 hour. The plates were washed 3 times with DELFIA wash buffer and incubated for 30 minutes with 20 μl of enhancement solution (Perkin Elmer 1244-105). Time-resolved fluorescence was then measured at 3240 and 615 nm using an EnVision® plate reader spectrophotometer (Perkin Elmer) according to the manufacturer's instructions. Chimeric anti-CD3-GUCY2c bispecific antibodies were screened for binding activity against recombinant human GUCY2c and human CD3 epsilon by ELISA. The top 6 clones showed dual binding activity in the low or sub-nanomolar range (Table 13).

Table 13

Clone Human EC ₅₀ GUCY2C (nM) EC ₅₀ human CD3 (nM) EC ₅₀ bispecific binding (nM) EC ₅₀ negative control (nM) GUCY2C-0074 >100 1.128 4,051 N.B. GUCY2C-0077 27.53 0.8873 7,504 N.B. GUCY2C-0078 23.51 1,446 9.68 N.B. GUCY2C-0098 60.19 7,244 185.6 N.B. GUCY2C-0104 >100 0.4179 2,479 N.B. GUCY2C-0105 79.76 0.3547 5.045 N.B.

Example 7: Bispecific Antibody-Mediated T Cell Activity

Human PBMC were isolated from healthy donor blood using Histopaque-177 (Sigma). Naïve T cells were isolated from PBMCs using a T cell enrichment kit from Stem Cell Technologies (T cell negative selection). GUCY2c-expressing human T84 tumor cells transfected with a luciferase expression construct were resuspended in R10 medium (RPMI, 10% FBS, 1% penicillin/streptomycin, 3 ml 45% glucose). T cells were also resuspended in R10 media and added to T84 cells at an effector to target ratio (E:T ratio) of 10:1 or 5:1. Cells were treated with serial dilutions of anti-GUCY2c bispecific antibody or a negative control anti-CD3 bispecific antibody and incubated at 37°C for 48 hours, followed by measurement of the luciferase signal using Neolite reagent (Perkin Elmer) using a Victor device (Perkin Elmer). EC ₅₀ values were calculated using Graphpad PRISM using four-parameter nonlinear regression analysis. Target-negative HCT116 or HT29 cells did not exhibit any anti-GUCY2c bispecific antibody-mediated cytotoxicity. The results of cell killing assays using chimeric anti-CD3-GUCY2c bispecific antibodies (E:T ratio=10:1) are shown in Table 14. All anti-CD3-GUCY2c bispecific antibodies demonstrated specific and potent T cell-dependent killing of GUCY2c-positive cells.

Table 14

Clone EC ₅₀ T84 tumor cells (nM) GUCY2C-0074 2.6 GUCY2C-0077 28.7 GUCY2C-0078 7.8 GUCY2C-0098 0.41 GUCY2C-0104 14.9 GUCY2C-0105 7.51

The results of cell killing assays using humanized anti-CD3-GUCY2c bispecific antibodies (E:T ratio=5:1) are shown in Table 15. All bispecific antibodies exhibited T cell-dependent cytotoxicity against GUCY2c-positive cells.

Table 15

Clone EC ₅₀ T84 tumor cells (nM) GUCY2C-0240 0.53 GUCY2C-0247 0.07 GUCY2C-1186 0.22 GUCY2C-1467 0.2 GUCY2C-1476 0.07 GUCY2C-1478 0.24 GUCY2C-1481 1.57 GUCY2C-1512 0.57 GUCY2C-1518 0.37 GUCY2C-1526 0.13 GUCY2C-1538 0.05 GUCY2C-1554 12.6 GUCY2C-1555 1.13 GUCY2C-1556 5.04 GUCY2C-1557 0.22 GUCY2C-1590 11.07 GUCY2C-1592 2.6 GUCY2C-1608 0.19

Example 8: Epitope Binning with Octet

The bispecific proteins GUCY2C-0074, -0077, -0104, -0105 and -0098 were assessed for competitive and non-competitive binding to human GUCY2c (SEQ ID NO: 224) using a tandem binning assay with OctetRED 384 (ForteBio). The Octet assay was performed at room temperature. Biotinylated human GUCY2c was diluted to 10 μg/ml in phosphate-buffered saline (PBS) without calcium and magnesium and captured onto streptavidin (SA) biosensors (18-5020, ForteBio) according to the manufacturer's instructions. Bispecific proteins were diluted to 300 nM in PBS. GUCY2c-coated biosensors were immersed in the first bispecific protein for 300 seconds, then immersed in the second bispecific protein for 300 seconds. Each of the bispecific antibodies was tested in this pairwise combinatorial manner. Bispecific antibodies competing for the same binding region on human GUCY2c were grouped in the same bin. By epitope binning of chimeric bispecific anti-CD3-GUCY2c antibodies using Octet, unique groups of epitopes recognized by anti-GUCY2c binding domains were identified (Table 16). The “+” symbol indicates competing epitopes; the symbol "+/-" indicates partially overlapping epitopes; the symbol "-" indicates non-competing epitopes; and “NT” indicates that no comparative analysis was performed.

Table 16

Clone GUCY2C-0074 + GUCY2C-0077 NT + GUCY2C-0098 + +/- + GUCY2C-0104 NT + +/- + GUCY2C-0105 NT + - + + GUCY2C-0074 GUCY2C-0077 GUCY2C-0098 GUCY2C-0104 GUCY2C-0105 Clone

Example 9: Humanization of Anti-GUCY2c Mouse Antibody Binding Domain

Humanization of the mouse anti-GUCY2c antibody variable regions was accomplished using a CDR grafting strategy. Complementary DNA (cDNA) containing the human acceptor framework, VH3-7 (SEQ ID NO: 176) for the heavy chain and VK1-39 (SEQ ID NO: 180) for the light chain, with complementarity determining region (CDR) donor sequences against GUCY2c were synthesized in vectors containing the human IgG1-3m constant region or the human kappa chain in the case of the light chain. The heavy chain vector consisted of a CH1 region (SEQ ID NO: 185), a hinge region (SEQ ID NO: 186) and a CH2-CH3 region including mutations affecting effector function, L234A, L235A and G237A (SEQ ID NO: 187; numbering according to the EU index). The light chain vector consisted of a kappa CL domain (SEQ ID NO: 184). Back mutations were inserted into the mouse sequence in the framework regions of the VH and VL regions and assessed for complete restoration of binding activity. Alignments of the VH and VL regions of the human acceptor framework, mouse parental clones, and fully active humanized variants are shown in Tables 17A, 17B, 18A, and 18B below for clone GUCY2C-0098.

Back mutations into the mouse sequence in the framework regions of the VH and VL regions were inserted and assessed for complete restoration of binding activity. Alignments of the VH and VL regions of the human acceptor framework, the mouse parental clones, and the fully active humanized variant GUCY2C-0241 are shown in Tables 17A and 17B below for clone GUCY2C-0098.

Table 17A

Clone HFW1 VHCDR1 HFW2 huIGHV3-7 EVQLVESGGGLVQPGGSLRLSCAAS
(SEQ ID NO: 5) GFTFSSYWMS (SEQ ID NO: 177) (ABM);
SYWMS (SEQ ID NO: 271) Kabat WVRQAPGKGLEWVA (SEQ ID NO: 6) GUCY2C-0098 QVQLQQPGAELVKPGASVKLSCKAS
(SEQ ID NO: 30) GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) WVKQRPGQGLEWIG (SEQ ID NO: 16) GUCY2C-0241 EVQLVESGGGLVQPGGSLRLSCAAS
(SEQ ID NO: 5) GYTFTSYWMH (SEQ ID NO: 27) (ABM);
SYWMH (SEQ ID NO: 259) (Kabat) WVRQAPGKGLEW IG (SEQ ID NO: 49)

Table 17B

Clone CDR2 VH HFW3 CDR3 VH huIGHV3-7 NIKQDGSEKYYVDSVKG(SEQ ID NO: 178) (Kabat);
KQDGSE (SEQ ID NO: 272) Chothia RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR(SEQ ID NO: 7) No GUCY2C-0098 EIKPSNGLTNYIEKFKN(SEQ ID NO: 28)(Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) KATLTVDKSATTAYMQLSSLTAEDSAVYYCTR(SEQ ID NO: 31) TITTTEGYWFFDV
(SEQ ID NO: 29) GUCY2C-0241 EIKPSNGLTNYIEKFKN(SEQ ID NO: 28) (Kabat);
KPSNGL (SEQ ID NO: 264) (Chothia) RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR(SEQ ID NO: 61) TITTTEGYWFFDV
(SEQ ID NO: 29)

Alignments of the VL region of the human acceptor scaffold, the mouse parental clone, and the fully humanized variant are shown in Tables 18A and 18B for clone GUCY2C-0098. CDRs are underlined when numbering according to Kabat. Bold indicates the original non-human germline amino acids present in the fully humanized clone.

Table 18A

Clone LFW1 VLCDR1 LFW2 huIGKV1-39 DIQMTQSPSSLSSASVGDRVTITTC
(SEQ ID NO: 80) RASQSISSYLN
(SEQ ID NO: 181) WYQQKPGKAPKLLIY
(SEQ ID NO: 81) GICU2C-0098 DIVLTQSPASLAVSLGQRATISC
(SEQ ID NO: 96) RASESVDYYGTSLMQ
(SEQ ID NO: 107) WYQQKPGQPPKLLIY
(SEQ ID NO: 110) GICU2C-0241 DIQ L TQSPSSLSSASVGDRVTITTC
(SEQ ID NO: 126) RASESVDYYGTSLMQ
(SEQ ID NO: 107) WYQQKPGKAPKLLIY
(SEQ ID NO: 81)

Table 18B

Clone VLCDR2 LFW3 CDR3 VL huIGKV1-39 AASSLQS
(SEQ ID NO: 182) GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
(SEQ ID NO: 82) QQSYSTPYT
(SEQ ID NO: 183) GICU2C-0098 AASNVES
(SEQ ID NO: 108) GVPARFSGSGSGTDFSLNIHPVEEDDIAMYFC
(SEQ ID NO: 111) QQTRKVYT
(SEQ ID NO: 109) GICU2C-0241 AAS NVE S
(SEQ ID NO: 108) GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
(SEQ ID NO: 82) QQ TRKV YT (SEQ ID NO: 109)

Table 19 shows that humanized GUCY2c binding domains as in human IgG1 retained binding to recombinant GUCY2c protein by ELISA.

Table 19

Clone Direct binding of huGUCY2C (nM) GUCY2C-0166 0.06935 GUCY2C-0179 5.143 GUCY2C-0193 0.2297 GUCY2C-0210 8,961 GUCY2C-0212 0.7694 GUCY2C-0241 0.1605

Table 20 shows that the humanized GUCY2c binding domains in both the anti-CD3 bispecific antibodies retained simultaneous binding to recombinant human GUCY2c and human CD3 proteins. GUCY2C-0240 is a humanized version of the anti-CD3 IgG bispecific antibody GUCY2C-0166. GUCY2C-0247 and GUCY2C-0250 are humanized bispecific anti-CD3 antibodies GUCY2C-0241 with different anti-CD3 variable regions.

Table 20

Clone huCD3-huGUCY2C in sandwich ELISA (nM) GUCY2C-0240 0.07992 GUCY2C-0247 0.705 GUCY2C-0250 76.9

Example 10: Further Humanization and Optimization of GUCY2C-Binding Domains Using Phage Display

In some embodiments, the substitution is a human germline substitution, in which the CDR residue is replaced with a corresponding human germline residue to increase the human amino acid content and reduce the potential immunogenicity of the antibody. For example, if the human germline VH3-7 backbone and the example antibody GUCY2C-0241 are used, the CDR1 alignment of the VH antibody GUCY2C-0241 (SEQ ID NO: 27) and human germline VH3-7 is shown in Table 21 (using the Kabat numbering system with determination of AbM CDR1 VH):

Table 21

Position 26 27 28 29 thirty 31 32 33 34 35 VH3-7 human germline G F T F S S Y W M S GUGY2C-0241 G Y T F T S Y W M H

In the case of positions 26, 28, 29, 31, 32, 33 and 34, the human germline residue and the corresponding residue of GUCY2C-0241 are the same, and germline substitution is not possible. In the case of positions 27, 30 and 35 (in bold and underlined), the human germline residue and the corresponding GUCY2C-0241 residue are different. The GUCY2C-0241 residues at these positions can be replaced with the corresponding human germline VH3-7 residue to further increase the human residue content.

The antibody or antigen binding fragment thereof may comprise a VH framework containing a human germline VH framework sequence. The VH framework sequence can be derived from human germline VH3, germline VH1, germline VH5, or germline VH4. Preferred human germline heavy chain scaffolds are those derived from germline VH1, VH3 or VH5. For example, VH scaffolds from the following germlines can be used: IGHV3-23, IGHV3-7, or IGHV1-69 (germline names based on the IMGT germline definition). Preferred human germline light chain scaffolds are those derived from germline Vκ or Vλ. For example, VL scaffolds from the following germlines can be used: IGKV1-39 or IGKV3-20 (germline names are based on the IMGT germline definition). Alternatively or additionally, the framework sequence may be a human germline consensus framework sequence, such as a human Vλ1 consensus framework, a Vκ1 consensus sequence, a Vκ2 consensus sequence, a Vκ3 consensus sequence, a VH3 germline consensus sequence, a VH1 germline consensus sequence, a germline consensus sequence VH5 lineage or VH4 germline consensus sequence. Human germline scaffold sequences are available in various public databases such as V-base, IMGT, NCBI or Abysis.

The antibody or antigen binding fragment thereof may comprise a VL framework containing a human germline VL framework sequence. The VL scaffold may contain one or more amino acid substitutions, additions or deletions while maintaining functional and structural similarity to the germ line from which it is derived. In some aspects, the VL frame is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the human germline VL framework sequence. In some aspects, the antibody or antigen binding fragment thereof comprises a VL framework comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid substitutions, additions or deletions relative to a human germline VL framework sequence.

The human germline VL scaffold may be a DPK9 scaffold (IMGT name: IGKV1-39). The human germline VL scaffold may be a DPK12 scaffold (IMGT name: IGKV2D-29). The human germline VL scaffold may be a DPK18 scaffold (IMGT name: IGKV2-30). The human germline VL scaffold may be a DPK24 scaffold (IMGT name: IGKV4-1). The human germline VL scaffold may be the HK102_V1 scaffold (IMGT name: IGKV1-5). The human germline VL scaffold may be a DPK1 scaffold (IMGT name: IGKV1-33). The human germline VL scaffold may be a DPK8 scaffold (IMGT name: IGKV1-9). The human germline VL scaffold may be a DPK3 scaffold (IMGT name: IGKV1-6). The human germline VL scaffold may be a DPK21 scaffold (IMGT name: IGKV3-15). The human germline VL scaffold may be the Vg_38K scaffold (IMGT name: IGKV3-11). The human germline VL scaffold may be a DPK22 scaffold (IMGT name: IGKV3-20). The human germline VL scaffold may be a DPK15 scaffold (IMGT name: IGKV2-28). The human germline VL scaffold may be the DPL16 scaffold (IMGT name: IGLV3-19). The human germline VL scaffold may be a DPL8 scaffold (IMGT name: IGLV1-40). The human germline VL scaffold may be the V1-22 scaffold (IMGT name: IGLV6-57). The human germline VL framework may be a consensus sequence of the human Vλ framework. The human germline VL framework may be a consensus sequence of the human Vλ1 framework. The human germline VL framework may be a consensus sequence of the human Vλ3 framework. The human germline VL framework may be a consensus sequence of the human Vκ framework. The human germline VL framework may be a consensus sequence of the human Vκ1 framework. The human germline VL framework may be a consensus sequence of the human Vκ2 framework. The human germline VL framework may be a consensus sequence of the human Vκ3 framework.

Determination of anti-GUCY2c antibody paratopes by extended binary substitution

A method for identifying critical CDR residues has been described that selects functional antibody variants from a library containing a human germline or corresponding rodent residue at each CDR position except CDR-H3 (Townsend et al., 2015, Proc. Natl. Acad. Sci USA 112(50):15354-15359). A scFv phage library was constructed in which all CDR positions except the human germline VH GUCY2C-0241 CDR3 (VH3-7/VK1-39) were randomized such that approximately 50% of the clones encoded the mouse amino acid GUCY2C-0241 and approximately 50% encoded a human germline amino acid at this position (Tables 17A, 17B, 18A and 18B). Libraries were prepared and subjected to 3-4 rounds of selection for human GUCY2c (SEQ ID NO: 232) as described below. Clones that retained binding to GUCY2c were isolated and their sequences determined.

In this experiment, positions at which human sequence was observed in less than approximately 20% of the binding clones were defined as essential mouse residues. Mouse residues were predominantly conserved at 25 positions (heavy chain residues H35, E50, P52a, S53, N54, G55, L56, T57, N58, I60, E61, K62, F63, and N65; light chain residues E27, V29, D30, L32, M33, Q34, N53, E55, T91, R92, and K93 according to the Kabat numbering system, and the AbM definition was used for CDR1 VH; the Chothia numbering system was used for CDR1 VL), suggesting that replacement of these residues with human germline residues is extremely unfavorable. Mouse and human residues were found at similar frequencies (defined as more than 20% human residue content) at heavy chain residues 27 (F/Y) and 30 (S/T) and light chain residues 54 (L/V) and 94 (A/ V; according to the Kabat numbering system; the AbM definition was used for CDR1 VH), indicating that the mouse sequence at these positions is not critical.

In addition, the extended binary replacement strategy included framework (FW) residues previously requiring back mutation into mouse amino acids (heavy chain positions I48 and G49 according to Kabat numbering). After phage selection and screening, the human amino acid sequence for heavy chain backbone position 48 was determined in approximately 10% of cases, suggesting that back mutation is favored. Heavy chain position 49 contained a mouse amino acid residue in 100% of cases, suggesting that this back mutation is necessary. The frequency of mouse or human amino acid sequence at the CDR or FW positions after extended binary substitution is shown in Table 22. Table 22 shows the frequency of inclusion of human amino acids at the GUCY2C-0098 CDR positions tested by extended binary substitution (using the Kabat numbering system, and for CDR1 VH used the AbM definition; for CDR1 VL the Chothia numbering system was used).

Table 22

Plot Mouse amino acids GUCY2C-0098 Human germline amino acids % human remains in source library % human residues in linking clones H27 Y F 50 21 H30 T S 35 28 H35 H S 39 2 H50 E N thirty 0 H52A P Q 46 0 H53 S D 33 0 H54 N G 37 0 H55 G S 41 1 H56 L E 39 0 H57 T K 46 0 H58 N Y 37 1 H60 I V 43 2 H61 E D 48 3 H62 K S 41 2 H63 F V 35 5 H65 N G 26 4 L27 E Q 48 12 L29 V I 46 4 L30 D S 28 2 L32 L Y 48 0 L33 M L 35 13 L34 Q N 20 6 L53 N S 20 2 L54 V L 50 26 L55 E Q 35 10 L91 T S 43 4 L92 R Y 57 2 L93 K S 48 10 L94 V T 46 24 H48 I V 26 10 H49 G A 48 0

Further optimization of binding domains after GUCY2c

Anti-GUCY2c antibody variants with improved stability were isolated by the following methods. The VH and VL regions of the humanized clone GUCY2C-0241 (SEQ ID NOs: 60 and 137) were synthesized as a single chain variable fragment (scFv) in a phage expression vector containing C-terminal His6 and c-myc tags (Blue Heron). Random mutations were introduced into the VH and VL CDR domains through NNK soft randomization (methods previously described in US Pat. No. 9884921). Oligonucleotides designed to selectively mutate one CDR residue at a time were synthesized by the supplier (IDT) and then used to make changes to the nucleic acid template by previously described methods (Townsend et al., 2015, Proc. Natl. Acad. Sci. USA 112(50):15354-15359). The oligonucleotide design also includes a mutation designed to eliminate a potential post-translational modification site (G55E heavy chain), which removes a potential deamidation site for asparagine NG (Chelius et al., Anal. Chem. 77(18):6004-6011, 2005) . Mutant clones were selected and submitted for sequencing to estimate mutation frequency. These phage expression vectors containing the mutagenized VH and VL CDR variants GUCY2C-0241 (SEQ ID NOs: 60 and 137) were then pooled into small sub-libraries for phage display.

Recombinant human, cynomolgus and mouse GUCY2c proteins were used in the selection and subsequent screening steps (SEQ ID NOs: 230, 232, 234, 236, 238 and 240). For selection involving biotinylated proteins, aliquots of the phage pools described above and streptavidin magnetic beads (Dynabeads M-280 with streptavidin) were blocked separately in 3% milk/PBS for 1 hour at room temperature, 60°C, or 65°C on a rotator. mixer (20 rpm). After incubation at different temperatures, the blocked phage aggregates were centrifuged at 4000 rpm for 10 minutes, the supernatants were transferred to new tubes and mixed with 6% blocking buffer. Blocked phages were incubated with excess molar concentrations of deselection reagents (i.e., target-free streptavidin particles containing dsDNA, insulin, and/or membrane extract reagents), incubated at room temperature for 1 hour on a rotary shaker (20 rpm ), were mixed with blocked magnetic particles with streptavidin and incubated for another hour. The deselected library was collected by particle deposition using a magnetic separator. Using a Kingfisher magnetic purification device (ThermoFisher, Springfield NJ, USA), biotinylated selection antigen (at various concentrations shown in Table 23) was incubated with the deselected phage library in a 96-well deep well plate for 1 hour at room temperature with periodic mixing. Particles were separated using a magnetic separator on a Kingfisher device and washed 4 times with PBS/0.1% Tween20 and 3 times with PBS in separate 96-well deep well plates. Bound phage was eluted by incubation with 100 mM triethylamine (TEA) for 8 minutes in a new 96-well plate followed by separation from magnetic beads. The eluted phage was neutralized with 2 M Tris-HCl, pH 7.5. An example of phage selection conditions for optimizing the binding domain of an anti-GUCY2c antibody is shown in Table 23. Table 23 also shows selection conditions used for phage display. "Dslxn" means the deselection conditions used. SA=streptavidin; D=DNA; I=insulin; M=membrane extract proteins. "Therm" corresponds to the thermal incubation performed before incubation with the target.

Table 23

Round Conditions R1 Group C1R1 Target huGUCY2C Target concentration 10 nM Dslxn SA, D, I, M Therm No R2 Group C1R2A Target cyGUCY2C Target concentration 1 nM Dslxn SA, D, I, M Therm 60°C R3 Group C1R3A1 Target huGUCY2C Target concentration 0.1 nM Dslxn SA, D, I, M Therm 65°C

The eluted phage was used to infect a 10 ml culture of E. coli ER2738 grown to mid-log phase (corresponding to OD ₆₀₀ ~0.5). Bacteria were infected with phage for 1 hour at 37°C with shaking at 150 rpm, concentrated after a centrifugation step and plated on 2× YT bioassay agar plates containing 2% glucose and 100 μg/ml ampicillin (2 -multiple YTAG). To determine phage titers, various dilutions of E. coli cultures infected with the input or output phage were also plated on 2× YTAG agar. After overnight growth at 30°C, 10 ml of 2× YTAG medium was added to each assay plate and the cells were resuspended by scraping off the bacterial lawn. Glycerol was added to this cell suspension to achieve a final concentration of 17% and stored in aliquots at -80°C until further use. To “rescue” the phage for the next round of selection, 100 µl of this cell suspension was used to inoculate 20 ml of 2× YTAG medium, grown at 37°C (300 rpm) to an OD ₆₀₀ of 0.3-0.5. Cells were then superinfected with 3.3 μl of helper phage MK13K07 and incubated at 37°C (150 rpm) for 1 hour. The cells were then centrifuged and the pellet was resuspended in kanamycin-containing/glucose-free medium (2× YT with 50 μg/ml kanamycin and 100 μg/ml ampicillin). This culture was grown overnight at 30°C (300 rpm). The phage was collected in the supernatant after centrifugation and was ready for use in the next round of selection as described above.

Preparation of raw periplasmic material for use in ELISA

ScFv can be expressed on the surface of the phage particle or in solution in the bacterial periplasmic space, depending on the growth conditions used. To induce scFv release into the periplasm, 96-well deep-well plates containing 2× YT media with 0.1% glucose/100 μg/ml ampicillin were inoculated with material from thawed glycerol stocks (one clone per well) using a - QPix colony picker (Molecular Devices) and grown at 37°C (999 rpm) for ∼4 h. Cultures were induced using IPTG at a final concentration of 0.02 mM and grown overnight at 30°C (999 rpm). Bacterial periplasmic contents (PeriPreps) were released using osmotic shock. Briefly, plates were centrifuged and pellets were resuspended in 150 μl of TES periplasmic buffer (50 mM Tris, 1 mM EDTA, 20% sucrose, pH7.4) followed by the addition of 150 μl of 1:5 TES:water and incubated on ice for 30 minutes. The plates were centrifuged for 20 minutes at 4000 rpm and the scFv-containing supernatant was collected.

Competitive ELISA

ELISA plates (NUNC-Maxisorb 460372) were coated with 1 μg/ml of the antigen of interest in PBS buffer overnight at 4°C. The coating solution was discarded and the plates were then blocked at room temperature for 2 hours with 70 μl PBS-3% BSA per well. The blocking solution was discarded and 20 µl of primary antibody was added, which was a mixture of the unknown sample as PeriPreps or purified protein (at a predetermined dilution, typically 1:5) and pure parent antibody at the desired concentration (typically EC ₅₀ -EC ₈₀ ). The plates were incubated for 2 hours at room temperature with slow shaking. Plates were washed 5 times with 100 μl per well wash buffer (Perkin Elmer 1244-114) and incubated for 1 hour with 20 μl of secondary antibody europium conjugate diluted 1:1000 in Delfia assay buffer (Perkin Elmer 1244-111) for streptavidin -Eu or anti-human IgG diluted 1:500 in the case of anti-mouse IgG. The plates were washed 5 times as described above and incubated for 30 minutes with 20 μl of enhancement solution (Perkin Elmer 1244-105). Time-resolved fluorescence was then measured at 320 and 615 nm using an EnVision® plate reader spectrophotometer (EnVision® multichannel plate reader spectrophotometer, Perkin Elmer) according to the manufacturer's instructions.

Conversion of ScFv to scFv-Fc, IgG and bispecific antibody

An ScFv exhibiting the desired properties was selected for subcloning into scFv-Fc, where the human IgG1 Fc domain was fused to the 3' end of the scFv. Briefly, fragments were amplified by standard polymerase chain reaction (PCR) with primers BssHII for 5' amplification and BclI for 3' amplification. Fragments were cut with appropriate restriction enzymes according to the manufacturer's instructions (New England Biolabs). Anti-GUCY2c scFv-Fc amplicons were purified from the gel (Qiagen Gel Purification Kit) and ligated into a pre-digested Pfizer proprietary mammalian expression vector containing the human IgG1 Fc domain. ScFv-Fc constructs were transformed into E. coli and sequenced as described above. Sequence verified expression vectors were then used for transient expression in HEK 293 mammalian cells as described above. Samples were purified using protein A affinity chromatography followed by buffer exchange to PBS as described above. Purified samples were then screened for recombinant GUCY2c protein binding activity and stability. An example of a scFv-Fc sequence is GUCY2C-0405 (SEQ ID NO: 213). This format consists of scFvs in a VH-VL orientation connected by linker 4, which is a glycine-serine linker (SEQ ID NO: 193). VH-VL is connected to a modified human IgG1 Fc domain (SEQ ID NO: 187) using two linker sequences, linker 5 (SEQ ID NO: 194) and linker 6 (SEQ ID NO: 195).

The optimized scFvs were also subcloned into human IgG1 and T cell bispecific formats as described above. Primers specific for variable region genes were designed for ITA cloning as described previously.

Example 11: Surface Plasmon Resonance of Optimized Anti-GUCY2c Antibodies

Optimized anti-GUCY2c antibodies were assessed for binding by surface plasmon resonance using a BIACORE™ 8K device (GE Healthcare). Using the CM5 sensor chip, anti-human Fc antibody (GE BR-1008-39) was first applied according to the manufacturer's instructions. Human anti-GUCY2c IgG was poured onto the chip for 30 sec at 0.5 μg/ml, 10 μl/min in Hanks' buffered saline (HBS) -EP+ pH=7.4. Human GUCY2c protein (SEQ ID NO: 224) at concentrations of 450 nM, 150 nM and 50 nM was then allowed to associate by flooding the chip for 54 sec at 50 μl/min and then dissociated for 300 sec. The chip was regenerated after each run with 3x 3M MgCl ₂ for 30 sec at 5 BIACORE™. The binding affinities ( _KD , nM) of optimized anti-GUCY2c binding domains in human IgG1 format to recombinant human GUCY2c protein are shown in Table 24. Several optimized anti-GUCY2c binding domains, such as GUCY2C-0486 (SEQ ID NOs: 214 and 215), demonstrated improved binding affinity compared to the parent, non-optimized binding domain (GUCY2C-0241).

Table 24

Clone K _D huGUCY2C (nM) GUCY2C-0210 85.67±2.76 GUCY2C-0212 25.98±0.01 GUCY2C-0241 9.81 GUCY2C-0456 13.75 GUCY2C-0478 29.85±0.26 GUCY2C-0483 6.92±0.13 GUCY2C-0486 5.47±0.07

Example 12: Binding Analysis by Thermal Test

To evaluate the improved thermal stability of the optimized variants, periplasmic preparations of the optimized phage clones (described above) or purified proteins were assessed for binding activity after thermal testing at elevated temperatures in a PCR apparatus. Greiner bio-one 384-well white ELISA plates (781074) were coated with 1 μg/mL target protein coating buffer (25 mM Na ₂ CO ₃ , 75 mM NaHCO ₃ , pH 9.6) O/N at 4°C. The plates were washed 3 times with PBS+0.05% TWEEN20 and then blocked for 1 hour with shaking at RT with blocking buffer (PBS+3% milk). Separately, plates were prepared for dilution of periplasmic drugs or purified protein by binding 1:10 periplasmic drugs or 1 μg/ml (EC ₈₀ ) of recombinant protein at room temperature in blocking buffer and 100 μl was transferred to a 96-well PCR plate. The PCR plate containing 100 μL of the diluted sample was then heated at the desired temperatures (typically 55°C to 75°C) for 30 minutes. The plates were allowed to cool for 15 minutes at room temperature and then centrifuged at 4000 rpm for 10 minutes. Blocked ELISA plates were emptied and 20 μl of heat-tested protein supernatant per well was added, then incubated for 1 h, vortexed at RT. The plates were washed 3 times as described above, and 20 μl of anti-6xHIS-Europium protein reagent (Perkin Elmer EU-N1) or anti-human IgG-Europium conjugate protein reagent (Perkin Elmer EU-N1) diluted 1:1000 in assay was added. DELFIA buffer (Perkin Elmer 1244-330) per well and incubated at RT for 1 hour with shaking. The plates were washed 3 times with 1× DELFIA wash solution (Perkin Elmer 1244-114), and 20 μl of enhancement solution (Perkin Elmer 1244-105) was added to each well and incubated for 30 minutes. The plates were read on an Envision plate reader for time-resolved fluorescence determination, first at OD320, then at OD615 according to the manufacturer's instructions. The temperature at which 50% activity was maintained was recorded as T50. In a thermal binding stability assay, the optimized GUCY2c-binding domains as scFv-Fc observed improved thermal stability after incubation at elevated temperatures compared to the non-optimized parental clone GUCY2C-0241 (designated GUCY2C-0295 in scFv-Fc format) (Tables 25A and 25B ).

Tables 25A and 25B show time-resolved fluorescence (TRF) for optimized GUCY2c-binding domains (as scFv-Fc proteins) at elevated temperature. The T50 value represents the temperature at which the clone retains 50% binding activity.

Table 25A

Clone 50°C 50.5°C 51.7°C 53.2°C 55.5°C 58.4°C 61.8°C GUCY2C-0295 113537.5 68863 40896 27068 21936 21409.5 21197 GUCY2C-0389 497589.5 517068 495444.5 397372 464052 150312.5 78680 GUCY2C-0393 477566.5 511271 501945.5 480373.5 451437.5 77154.5 3630.5 GUCY2C-0401 506357.5 526131 535157.5 517779 523479 525247 469885.5 GUCY2C-0403 488139.5 544047.5 526733 536576 526171.5 522549.5 506987.5 GUCY2C-0411 471676.5 522040 542235 531706 539339 515333.5 425089 GUCY2C-0413 574068 598919 607913.5 603587.5 606227 590548 562273.5 GUCY2C-0418 547580 567189.5 526030.5 539879 526607 492907.5 407950 GUCY2C-0421 515013 487958.5 511088.5 510749 547231.5 529472 527035.5 GUCY2C-0423 411893.5 399052.5 462363.5 343803 464695.5 363302 8103 GUCY2C-0425 484807 479583 556277.5 510768 551751.5 548408 527384.5 GUCY2C-0427 512520 546980.5 547056 523887.5 537909.5 533628 498146 GUCY2C-0428 517436.5 541570.5 548392.5 536214.5 536255.5 523982.5 507148 GUCY2C-0435 487047.5 528185 517133.5 489329 490196.5 326153.5 42327 GUCY2C-0439 628296 568971 578626.5 576348.5 591356 585622 597929.5 GUCY2C-0445 536524 543057.5 522756 529305.5 529928 461742 36773.5 GUCY2C-0447 489262.5 499721 483027.5 484959 490929.5 470553.5 424560.5 GUCY2C-0448 565712.5 572475 566258 558343.5 560063 559518 547452 GUCY2C-0450 579746.5 595076.5 582737 578649.5 574037.5 563058.5 555110.5 GUCY2C-0451 564219.5 565212 541781 561199.5 534570 542623 528037

Table 25B

Clone 64.6°C 66.8°C 68.1°C 69.6°C 70°C T50 GUCY2C-0295 9853.5 1922 594 642.5 464.5 50.41 GUCY2C-0389 51040.5 21131.5 5766 2710.5 640.5 57.98 GUCY2C-0393 1677 909.5 2306 532 745.5 57.65 GUCY2C-0401 26531 3014.5 3061.5 2215 1178.5 62.77 GUCY2C-0403 498400.5 448403.5 298626.5 158161.5 77527 68.01 GUCY2C-0411 52317 20580 5945.5 1917 1162 62.44 GUCY2C-0413 573518.5 541790.5 469893 256224 108873 68.65 GUCY2C-0418 231526 136359 41704 7131.5 2812.5 62.23 GUCY2C-0421 489965 156593 1903.5 1135 980.5 66.44 GUCY2C-0423 2270.5 1542.5 785.5 879.5 929.5 59.23 GUCY2C-0425 494844.5 160062.5 2404.5 944 804 66.45 GUCY2C-0427 486978 224510 2906.5 1084 902.5 66.67 GUCY2C-0428 476739 96235.5 2133.5 1663.5 1332.5 66.1 GUCY2C-0435 17360 2504.5 1119.5 820.5 1061.5 58.65 GUCY2C-0439 562458.5 469494 249716.5 35294 6786.5 67.88 GUCY2C-0445 9384.5 1404.5 372.5 318.5 416 59.21 GUCY2C-0447 241324.5 12424.5 1271 391.5 1092.5 64.6 GUCY2C-0448 510974.5 221540 2645 724.5 648 66.62 GUCY2C-0450 518052.5 131373 1147 468.5 593 66.26 GUCY2C-0451 496304.5 189156 1814.5 693.5 731.5 66.53

The amino acid sequences of anti-GUCY2c antibody clones demonstrating improved thermal stability were compared to the parental clone GUCY2C-0241 (VH SEQ ID NO: 60 and VL SEQ ID NO: 137). Changes in amino acid content of the VH CDR resulting in improved stability are shown in Table 26A. Changes at positions 59, 60, 61, and 62 appear to have the greatest impact on stability, correlating with the largest changes in stability (according to the Kabat numbering system using the AbM definition for CDR1 VH).

Table 26A

Position Parent amino acid Optimized Amino Acid CDR1 VH thirty T G 31 S N 32 Y D CDR2 VH 59 Y V, I, L 60 I N, H, A, S, G 61 E D, P 62 K Q, R, T CDR3 VH 95 T L 96 I V 97 T L, M 98 T K 99 T P, K, A, S, R

Amino acid changes in CDR VL resulting in improved stability are shown in Table 26B (using the Chothia numbering system for CDR1 VL and the Kabat numbering system for CDR2 VL and CDR3). Changes at positions 53, 54, 55 in CDR2 VL and positions 93 and 94 in CDR3 VL appear to have the greatest impact on stability, correlating with the largest changes in stability.

Table 26B

Position Parent amino acid Optimized Amino Acid CDR1 VH 24 R T, K 30a Y W,A 30d T S, H, A, R, N 33 M L 34 Q H CDR2 VH 50 A G 51 A G 52 S T, H 53 N K, H 54 V L, R, I, G, K, T 55 E A, Y, W, V 56 S P, T, A CDR3 VH 89 Q M, L, N 91 T S, N 93 K R, N, Q, S 94 V A, E, G, D, I, Q 97 T S

Example 13: Recombinant Protein Binding by Optimized Anti-CD3-GUCY2c Bispecific Antibodies

The optimized anti-GUCY2c binding domains were assessed in a bispecific format for simultaneous binding of the recombinant protein to human GUCY2c (SEQ ID NO: 224) and human CD3 (SEQ ID NO: 242) at room temperature and 60°C by the DELFIA method described above. The optimized bispecific antibodies were also assessed for direct binding to human GUCY2c (SEQ ID NO: 232) and cynomolgus GUCY2c (SEQ ID NO: 236) at room temperature and 60°C by the DELFIA method described above. The dual binding of optimized anti-GUCY2c bispecific antibodies to human GUCY2c and human CD3 are shown in Table 27. Optimized anti-CD3-GUCY2c bispecific antibodies, such as GUCY2C-1478, showed strong binding at room temperature and 60°C.

Table 27

Clone DELFIA huCD3-huGUCY sandwich format at RT (1 µg/ml) DELFIA huCD3-huGUCY sandwich format at 60°C (1 µg/ml) GUCY2C-1186 536148 137679 GUCY2C-1467 830590 620774 GUCY2C-1476 860194.5 682977.5 GUCY2C-1478 833155.5 727935 GUCY2C-1481 790943.5 694815.5 GUCY2C-1512 800436 673743.5 GUCY2C-1518 430465 87434.5 GUCY2C-1526 568727.75 147975 GUCY2C-1527 405448.25 67409 GUCY2C-1538 617581 151603 GUCY2C-1554 138640 14584 GUCY2C-1555 353355.25 68362.25 GUCY2C-1556 217761.5 40731.75 GUCY2C-1557 473322 190626.25 GUCY2C-1590 165141 71515 GUCY2C-1591 422409.25 159723.25 GUCY2C-1592 262425.75 79415.75 GUCY2C-1608 523911.5 192704.5

The results of the direct binding assay of optimized anti-GUCY2c bispecific antibodies to human and cynomolgus GUCY2c are shown in Table 28. Optimized anti-CD3-GUCY2C bispecific antibodies, such as GUCY2C-1478, showed strong binding at room temperature to human and cynomolgus GUCY2c, but observed some loss of binding signal after incubation at 60°C.

Table 28

Clone EC ₅₀ huGUCY2C at RT (nM) EC ₅₀ CyGUCY2C at RT (nM) EC ₅₀ huGUCY2C at 60°C (nM) EC ₅₀ CyGUCY2C at 60°C (nM) GUCY2C-1478 4,387 4,902 19.24 15.65 GUCY2C-1481 7,836 8.34 27.36 64.99 GUCY2C-1554 18.32 13.93 N.B. N.B. GUCY2C-1555 5,391 6.85 39.44 94.86 GUCY2C-1556 13.58 10.14 65.58 62.41 GUCY2C-1557 6,382 7.51 38.2 53.52 GUCY2C-1590 20.25 13.31 48.65 57.3 GUCY2C-1592 13.38 14.39 94.09 75.97 GUCY2C-1608 8,775 9.835 23.67 49.87 Negative control N.B. N.B. N.B. N.B.

Example 14: Cell Binding of Anti-CD3-GUCY2c Bispecific Antibodies by Flow Cytometry

Optimized anti-CD3 bispecific antibodies were titrated for cell surface binding to GUCY2c on T84 cells and to human CD3 on naïve human T cells isolated from fresh human peripheral blood mononuclear cells (PBMC) by the flow cytometry methods described above. The humanized and optimized anti-GUCY2c clones GUCY2C-0247 and GUCY2C-1608 exhibited binding affinities to T84 cells expressing human GUCY2c in the low and subnanomolar range (Figures 2A and 2B). Humanized and optimized anti-GUCY2c bispecific antibodies were also assessed for binding to cell surface CD3 by flow cytometry (Figures 3A and 3B) and observed binding affinity to naïve human T cells in the low to mid nanomolar range.

Example 15: Reducing the Risk of Immunogenicity of Anti-GUCY2c Bispecific Antibodies

When certain therapeutic proteins are administered to patients, unwanted immune responses such as the formation of anti-drug antibodies (ADA) affect the effectiveness of the drug and raise safety concerns for the patient (Yin L., et al., Cell Immunol. Jun; 295 (2):118-26, 2015). ADAs can be classified into two groups: neutralizing ADAs (NAbs) or non-neutralizing ADAs (non-NAbs) depending on whether they inhibit the pharmacological activity of the therapeutic protein (Yin L., et al., Cell Immunol. Jun; 295(2) :118-26, 2015). There are two possible mechanisms by which NAbs and non-NAbs may contribute to decreased drug efficacy. First, NAbs directly block the binding of the therapeutic protein to the target molecule, thereby reducing its therapeutic efficacy (Yin L., et al., Cell Immunol. Jun; 295(2):118-26, 2015). Second, NAbs and non-NAbs may contribute to increased clearance by affecting the pharmacokinetics (PK) of TPP, thereby interfering with the effectiveness of the drug (Yin L., et al., Cell Immunol. Jun; 295(2): 118-26, 2015). Immunogenicity to a therapeutic protein can be generated through T cell-dependent and T cell-independent pathways. In the T cell-dependent pathway, T cells are activated by recognizing therapeutic protein-derived antigenic peptides presented by major histocompatibility complex class II (MHC II) molecules on antigen-presenting cells. The activated T cells then stimulate the B cells to produce ADAs specific for the therapeutic protein (Yin L., et al., Cell Immunol. Jun; 295(2):118-26, 2015). To minimize the potential for immunogenicity resulting from T cell-dependent responses, attempts have been made to reduce the antigenic potential of anti-CD3-GUCY2c bispecific antibody sequences by in silico methods described below.

Sequences were analyzed in two ways (detailed below) to identify potential T cell epitopes. Any sequence marked according to the rules presented herein for any method was considered an epitope. In the present description, sequences are examined primarily at the amino acid 9-mer level.

Method 1: Sequences are subjected to EpiMatrix analysis in the ISPRI software package (ISPRI v 1.8.0, EpiVax Inc., Providence, RI (2017); Schafer JRA, Jesdale BM, George JA, Kouttab NM, De Groot AS. Prediction of well-conserved HIV-1 ligands using a matrix-based algorithm, EpiMatrix. Vaccine 16(19), 1880-84, 1998). The raw results are a ranking of the binding probability of each 9-mer amino acid fragment relative to 8 different HLA types. Thus, there are 8 predictions ("observations") for each 9-mer. 9-mers are obtained starting from each individual linear numbering position of the sequence (thus the same 9-mer can occur more than once in the same sequence). If any 4 observations indicate that the 9-mer is in the top 5% of binders (meaning it is predicted to be in the top 5% of binders for at least 4 HLA types), the 9-mer is considered a predicted epitope ( "epitope"). Alternatively, if any 1 of 8 predictions indicate that the 9-mer is in the top 1% of binders, the 9-mer is also considered a predicted epitope.

Method 2: Sequences were analyzed using the MHC-II consensus binding method (Wang P, Sidney J, Kim Y, Sette A, Lund O, Nielsen M, Peters B. Peptide binding predictions for HLA DR, DP and DQ molecules. BMC Bioinformatics 11:568, 2010; Wang P, Sidney J, Dow C, Mothé B, Sette A, Peters B. A systematic assessment of MHC class II peptide binding predictions and evaluation of a consensus approach. PLoS Comput Biol. 4(4), e1000048, 2008in IEDB (Vita R, Overton JA, Greenbaum JA, Ponomarenko J, Clark JD, Cantrell JR, Wheeler DK, Gabbard JL, Hix D, Sette A, Peters B. The immune epitope database (IEDB) 3.0. Nucleic Acids Res. Jan 28 (43), D405-12, 2015; IEDB MHC-II Binding Predictions, http://www.iedb.org).

The software output categorizes the results into 15 measures. Consensus scores and percentile rankings for each 15-mer and HLA type combination are obtained. But the individual scores from which the consensus results for each 15-mer are derived are a ranking of some of the 9-mers found in the 15-mer: in each method used to obtain the consensus results, the report is provided as a percentile rank for the 9-mer in 15-measure, and the consensus score taken as the value for the overall 15-measure is a prediction for the 9-measure having the median scores. We have classified a 9-mer as an epitope if (a) it is selected as a consensus typical result for a 15-mer, and (b) it has a percentile rank in the top 10% of binders for the HLA type in question, and if criteria (a) and (b) matched for three or more distinct HLA types for the same 9-mer (i.e., three observations). The HLA types considered were DRB1*01, 1*03, 1*04, 1*07, 1*08, 1*11, 1*13, and 1*15, which are the same HLA types in the standard ISPRI/EpiMatrix report. Thus, although the primary output of the method is a ranking of 15-mers, the present inventors reinterpret the data to obtain a list of predicted 9-mer epitopes for ease of comparison with Method 1.

The present inventors have classified each epitope as a germline or non-germline epitope. In the case of antibodies, the present inventors further classified each epitope based on its location in the antibody (CDR or non-CDR). We filtered human V domain sequences obtained from IMGT (www.imgt.org) to remove germline annotations as pseudogenes or open reading frames (ORFs). Any predicted 9-mer epitope found in the remaining sequences was considered a germline epitope. Epitopes found in the J or C regions (including IgG1, IgG2, IgG3, and IgG4) or junctions between these regions were also classified as germline epitopes. In other words, the epitope is classified as a non-germline epitope. CDR definitions are based on the Kabat numbering system (Kabat EA, Wu TT, Perry HM, Gottesman KS, Foeller C. Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH Publication No. 91-3242, 1991), where CDRs are defined to include the following residues: CDR1 VH (H26-H35, including insertions such as H35A, up to but not including H36), CDR2 VH (H50-H65 inclusive), CDR3 VH (H95-H102 inclusive), CDR1 VL (L24-L34 inclusive), CDR2 VL (L54-L56, inclusive), CDR3 VL (L89-L97, inclusive). A predicted 9-mer epitope is a CDR epitope if any of these amino acids are part of the CDR region. It should be noted that the starting position (H26) chosen by the authors of the present invention for CDR1 VH differs from that described in some other publications using the Kabat annotation.

In the case of a single chain or pair of antibody VH and VL domains, total scores can be calculated by summing each of the 9-mers as follows. All individual combinations of 9-mer and HLA type (“cases”) are examined, regardless of whether the 9-mer is an epitope. If a particular observation indicates that a peptide is in the top 5% of binders for a specified HLA type, the EpiMatrix Z-score for that observation is added to the running sum associated with the entire protein sequence. The total number of observations studied is also recorded. The only exception is that all observations involving 9-mers identified by ISPRI as “T-regitopes” are considered to have an EpiMatrix score of zero. In the subtotal, the raw scores of 0.05*2.2248 are subtracted from each observation (including T-regitopes). Final scores are calculated as follows:

Adjusted T-reg scores = (subtotal)*1000/(number of observations)

Lower scores indicate lower predicted immunogenic potential.

Scores only include prediction using ISPRI/EpiMatrix and do not include information derived from IEDB. Thus, any strong HLA binders predicted by the IEDB, but not the ISPRI, do not contribute to the scores. In theory, sequences could contain multiple IEDB-predicted HLA binders and still have favorable adjusted T-reg scores unless the same sequences are also predicted as likely binders by EpiMatrix.

Germline alignment of unoptimized and optimized VH and VL regions against GUCY2c. Residues in bold represent T cell epitope hotspots determined by the in silico methods described above. In some embodiments, the substitution is a human germline substitution in which the CDR residue is replaced with a corresponding human germline residue to increase the human amino acid content and reduce the potential immunogenicity of the antibody (as described above). In other embodiments, the substitution is a different amino acid that reduces the potential immunogenicity of the antibody and does not interfere with the binding properties of the antibody.

For example, if the human germline VH3-7 framework is used with the example antibody GUCY2C-0241, then the CDR2 alignment of GUCY2C-0241 VH (SEQ ID NO: 28) and human germline VH3-7 (SEQ ID NO: 178) is as follows: presented in Table 29 below (using the Kabat numbering system):

Table 29

Clone 50 51 52 52 A 53 54 55 56 57 58 59 60 61 62 63 64 65 VH3-7 N I K Q D G S E K Y Y V D S V K G GUGY2c-0241 E I K P S N G L T N Y I E K F K N

In the case of positions 51, 52, 59 and 64, the human germline residue and the corresponding residue of GUCY2C-0241 are the same, and germline substitution is not possible. In the case of positions 50, 52A, 53, 54, 55, 56, 57, 58, 60, 61, 62, 63 and 65 (in bold), the human germline residue and the corresponding GUCY2C-0241 residue are different. When the GUCY2C-0241 sequence is subjected to in silico T cell epitope analysis, potential peptide sequences are identified (underlined). They are then further analyzed for non-human germline content with T cell epitope scores >10 and the non-human germline residues are used for mutagenesis (shown above in bold and underlined). Using soft randomization mutagenesis (methods previously described in US Pat. No. 9,884,921), these residues were mutagenized and tested for retention of binding activity and stability.

Example 16: T cell-mediated cell killing activity of optimized anti-CD3-GUCY2c bispecific antibodies

Optimized anti-CD3-GUCY2c bispecific antibodies were assessed for T cell-mediated cytotoxicity against tumor cells as previously described. Optimized bispecific antibodies against GUCY2c demonstrated improved effective T cell-mediated killing of T84 tumor cells. The results of cell killing assays using the humanized anti-CD3-GUCY2c bispecific antibody (E:T ratio=5:1) are shown in Table 30 and Figures 4A-4D.

Table 30

Clone EC ₅₀ T84 tumor cells (nM) GUCY2C-0240 0.53 GUCY2C-0453 17.65 GUCY2C-0454 14.34 GUCY2C-0247 0.07 GUCY2C-0381 2.7 GUCY2C-1186 0.22 GUCY2C-1467 0.2 GUCY2C-1476 0.07 GUCY2C-1478 0.24 GUCY2C-1481 1.57 GUCY2C-1512 0.57 GUCY2C-1518 0.37 GUCY2C-1526 0.13 GUCY2C-1538 0.05 GUCY2C-1554 12.6 GUCY2C-1555 1.13 GUCY2C-1556 5.04 GUCY2C-1557 0.22 GUCY2C-1590 11.07 GUCY2C-1592 2.6 GUCY2C-1608 0.19

Example 17: Differential Scanning Calorimetry (DSC) Analysis

Proteins were diluted in phosphate-buffered saline (PBS) to 0.6 mg/ml in a volume of 400 μl. PBS was used as a blank buffer in the reference cell. PBS contained 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na ₂ HPO ₄ and 1.47 mM KH ₂ PO ₄ , pH 7.2. Samples were dispensed into a MicroCal VP-Capillary DSC sample tray using an autosampler (Malvern Instruments Ltd, Malvern, UK). Samples were equilibrated for 5 minutes at 10°C and then brought to 110°C at a rate of 100°C per hour. The filtration period was chosen to be 16 seconds. The raw data were background corrected and protein concentrations were normalized. Origin 7.0 software (OriginLab Corporation, Northampton, MA) was used to fit the data to the MN2-State model with the appropriate number of transitions. Differential scanning calorimetry results of optimized binding domains against GUCY2c in scFv-Fc format are shown in Table 31. Optimized clones exhibit improved unfolding temperature (Tm1) compared to the parent, non-optimized clone GUCY2C-0247 (designated GUCY2C-0295 in scFv-Fc format) .

Table 31

Clone Tm1 DSC Tm2 DSC Tm3 DSC GUCY2C-0295 57.9 68.8 83.6 GUCY2C-0389 64.2 67.5 83.6 GUCY2C-0393 64.1 67.5 83.6 GUCY2C-0401 69.1 69.1 83.6 GUCY2C-0403 73.3 73.3 84.5 GUCY2C-0411 69.1 69.1 83.8 GUCY2C-0413 71.6 76.2 84.5 GUCY2C-0418 51 68.1 83.7 GUCY2C-0421 71.5 71.5 84 GUCY2C-0423 66.1 66.1 83.8 GUCY2C-0425 71.6 71.6 84 GUCY2C-0427 68.3 72.1 84 GUCY2C-0428 71.2 71.2 84.1 GUCY2C-0435 65.3 65.3 83.7 GUCY2C-0439 72.8 72.8 84.4 GUCY2C-0445 67 67 83.8 GUCY2C-0447 69.7 69.7 84.3 GUCY2C-0448 71.6 71.6 84.3 GUCY2C-0450 71.2 71.2 84.2 GUCY2C-0451 71.6 71.6 84.2

Example 18: Surface Plasmon Resonance (SPR) Analysis of Anti-CD3-GUCY2c Bispecific Antibodies

The binding affinities of anti-CD3-GUCY2c bispecific antibodies to human GUCY2c (SEQ ID NO: 224), cynomolgus GUCY2c (SEQ ID NO: 236) and mouse GUCY2C (SEQ ID NO: 240) were determined using the BIACORE™ device T200 (GE Healthcare) at 25°C and 37°C with a collection rate of 10 Hz. Human GUCY2c, cynomolgus GUCY2c, and mouse GUCY2c were directly immobilized onto the surface of three different flow cells of a CM5 sensor chip (BR100530, GE Healthcare) using an amino coupling kit (BR100050, GE Healthcare) according to the manufacturer's instructions. The final immobilization levels of human, cynomolgus, and mouse GUCY2c were 142 resonance units (RU), 61 RU, and 201 RU, respectively. Flow cell 1 was used as the reference flow cell. A series of threefold dilutions of anti-CD3-GUCY2c bispecific antibodies with concentrations ranging from 450 nM to 5.56 nM were injected onto the sensor surface over 52 seconds at a flow rate of 50 μL/min. Dissociation was monitored for 400 seconds and the surface was regenerated with 10 mM glycine, pH 2.1. The running and sample buffer were 10 mM HEPES, pH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.05% P-20 (HBS-EP+). Data were obtained in two parallels. Binding affinities and rate constants were determined by fitting the resulting sensogram data to a 1:1 Langmuir model using BIACORE™ T200 version 3.0 analysis software (GE Healthcare). The binding affinities of anti-GUCY2c bispecific antibodies to human CD3 (SEQ ID NO: 242) and cynomolgus CD3 (SEQ ID NO: 244) were determined using a BIACORE™ T200 device (GE Healthcare) at 25°C and 37°C with an acquisition rate of 10 Hz Human and cynomolgus CD3 were captured on the surface of three different flow cells of the CM5 sensor chip (BR100050, GE Healthcare) using the His-group capture kit (28995056, GE Healthcare) according to the manufacturer's instructions. Human CD3 (SEQ ID NO: 242) capture levels were 8 resonance units (RU) and 16 RU on flow cells 2 and 3, respectively, with 10 RU cynomolgus CD3 captured on flow cell 4. Flow cell 1 was used as reference . A series of threefold dilutions of anti-GUCy2c bispecific protein with concentrations ranging from 100 nM to 3.7 nM were injected onto the touch surface over 55 seconds at a flow rate of 50 μL/min. Dissociation was monitored for 300 seconds and the surface was regenerated with 10 mM glycine, pH 1.5. The running and sample buffer were 10 mM HEPES, pH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.05% P-20 (HBS-EP+). Data were obtained in two parallels. Binding affinities and rate constants were determined by fitting the resulting sensogram data to a 1:1 Langmuir model using BIACORE™ T200 version 3.0 analysis software (GE Healthcare). The results of the BIACORE™ anti-CD3-GUCY2c bispecific antibody binding assay to human, cynomolgus and mouse GUCY2c under various conditions, including assay orientation and temperature, are shown in Tables 32A-32J. Humanized and optimized GUCY2C-binding domains showed preserved or improved binding to GUCY2C and CD3 as bispecific antibodies. In some cases, a decrease in binding affinity was observed at elevated temperature (37°C).

Table 32A shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to human GUCY2c. The capture method used was capture with anti-human IgG antibody at 25°C.

Table 32A

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0074 4.41E+4±2.17E+3 2.00E-3±4.27E-5 45.40±2.90 4 GUCY2C-0098 1.36E+5±4.79E+1 6.75E-4±1.97E-6 4.96±0.02 2 GUCY2C-0240 9.62E+4±4.76E+3 3.64E-3±1.22E-5 37.90±1.75 2 GUCY2C-0247 9.82E+4±1.64E+3 4.57E-4±2.84E-6 4.65±0.09 4 GUCY2C-1478 1.18E+5±7.24E+2 4.34E-4±3.05E-5 3.69±0.28 2

Table 32B shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to human GUCY2c. The capture method used was direct immobilization at 25°C.

Table 32B

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0077 1.43E+5±1.7E+4 2.24E-2±1.00E-3 160.00±25.00 2 GUCY2C-0098 1.45E+5±2.41E+3 1.04E-3±2.61E-5 7.19±0.30 2 GUCY2C-0104 1.48E+5±1.00E+4 3.86E-3±2.00E-5 26.10±1.90 2 GUCY2C-0240 9.62E+4±4.76E+3 3.64E-3±1.22E-5 37.90±1.75 2 GUCY2C-0247 1.44E+5±1.83E+3 4.33E-4±2.88E-6 3.01±0.02 2 GUCY2C-1608 1.01E+5±2.19E+3 7.58E-4±1.68E-6 7.47±0.15 2

Table 32C shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to human GUCY2c. The capture method used was direct immobilization at 37°C.

Table 32C

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 2.51E+5±4.65E+3 3.77E-3±1.85E-5 15.02±0.35 2 GUCY2C-1608 1.61E+5±8.47E+3 2.17E-3±6.58E-5 13.52±1.12 2

Table 32D shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to cynomolgus GUCY2c. The capture method used was capture with anti-human IgG antibody at 25°C.

Table 32D

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0074 4.73E+4±7.70E+3 2.84E-3±3.60E-4 60.35±2.15 2 GUCY2C-0098 1.34E+5±1.63E+3 5.24E-4±3.93E-5 3.93±0.34 2 GUCY2C-0240 2.61E+4±7.10E+3 2.34E-3±5.24E-4 91.20±12.50 4 GUCY2C-0247 8.55E+4±1.35E+4 3.69E-4±4.35E-5 4.35±0.24 4

Table 32E shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to cynomolgus GUCY2c. The capture method used was direct immobilization at 25°C.

Table 32E

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 2.06E+5±1.17E+3 5.77E-4±3.50E-6 2.80±0.03 2 GUCY2C-1608 1.38E+5±1.04E+3 4.17E-4±7.61E-6 3.01±0.03 2

Table 32F shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to cynomolgus GUCY2c. The capture method used was direct immobilization at 37°C.

Table 32F

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 3.10E+5±7.75E+3 2.64E-3±3.76E-5 8.52±0.33 2 GUCY2C-1608 1.91E+5±1.18E+4 1.66E-3±4.67E-5 8.68±0.29 2

Table 32H shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to mouse GUCY2c. The capture method used was capture with anti-human IgG antibody at 25°C.

Table 32H

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0074 1.48E+4±5.32E+1 2.82E-3±1.08E-4 191.01±6.60 2 GUCY2C-0098 3.75E+4±1.31E+3 4.02E-3±2.25E-5 107.19±3.15 2 GUCY2C-0240 1.06E+4±8.30E+1 3.36E-3±6.89E-5 316.54±8.95 2

Table 32I shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to mouse GUCY2c. The capture method used was direct immobilization at 25°C.

Table 32I

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 1.34E+5±1.61E+3 2.68E-3±1.55E-5 19.97±0.36 2 GUCY2C-0240 1.22E+5±3.83E+3 5.15E-3±1.35E-5 42.34±1.44 2 GUCY2C-0247 1.50E+5±1.67E+3 1.83E-3±9.99E-6 12.22±0.07 2 GUCY2C-1608 9.12E+4±1.13E+3 1.57E-3±3.04E-6 17.27±0.25 2

Table 32J shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to mouse GUCY2c. The capture method used was direct immobilization at 37°C.

Table 32J

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 1.96E+5±1.25E+4 2.01E-2±1.48E-3 102.66±0.97 2 GUCY2C-1608 1.29E+5±7.10E+3 1.16E-2±1.61E-4 90.17±3.71 2

Tables 33A-33F show the BIACORE™ binding assay bispecific antibodies against CD3-GUCY2c with human and cynomolgus CD3 under different conditions, including assay orientation and temperature.

Table 33A shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to human CD3 protein. The capture method used was capture with anti-human IgG antibody at 25°C.

Table 33A

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0074 4.71E+5±2.16E+5 8.61E-4±1.56E-5 2.17±0.99 4 GUCY2C-0098 2.97E+5±1.28E+3 9.67E-4±2.15E-5 3.26±0.09 2 GUCY2C-0240 3.47E+5±8.83E+3 1.29E-3±4.5E-5 3.73±0.19 4 GUCY2C-0247 2.95E+5±2.17E+3 9.85E-4±6.34E-6 3.34±0.04 4

Table 33B shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to human CD3. The capture method used was capture with anti-His antibody at 25°C.

Table 33B

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 1.10E+6±2.14E+4 5.24E-3±8.88E-5 4.75±0.08 4 GUCY2C-1478 3.55E+5±6.10E+3 9.56E-3±1.37E-4 26.93±0.76 3 GUCY2C-1608 4.18E+5±2.78E+4 9.99E-3±2.41E-4 23.97±0.97 4

Table 33C shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to human CD3. The method used was capture with anti-His antibody at 25°C.

Table 33C

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 2.51E+6±6.34E+5 5.05E-2±1.43E-2 19.93±0.72 4 GUCY2C-1608 7.44E+5±2.31E+5 9.00E-2±2.18E-2 123.77±10.95 4

Table 33D shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to cynomolgus CD3. The method used was capture with anti-His antibody at 25°C.

Table 33D

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0074 4.25E+5±1.36E+4 1.12E-3±8.98E-5 2.64±0.28 4 GUCY2C-0098 2.84E+5±9.80E+3 1.01E-3±3.55E-5 3.56±0.25 2 GUCY2C-0240 4.40E+5±5.00E+2 2.07E-3±8.00E-5 4.71±0.19 2 GUCY2C-0247 2.80E+5±1.48E+4 1.00E-3±6.76E-5 3.61±0.43 4

Table 33E shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to cynomolgus CD3. The method used was capture with anti-His antibody at 25°C.

Table 33E

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 9.69E+5±1.85E+4 4.78E-3±1.03E-4 4.94±0.01 2 GUCY2C-1608 3.81E+5±2.33E+4 9.16E-3±2.31E-4 24.12±0.87 2

Table 33F shows the binding kinetics of anti-CD3-GUCY2c bispecific antibodies to cynomolgus CD3. The method used was capture with anti-His antibody at 25°C.

Table 33F

Clone ka (1/Ms) kd (1/s) K _D (nM) N GUCY2C-0098 2.28E+6±3.59E+5 4.42E-2±7.86E-3 19.34±0.40 2 GUCY2C-1608 5.14E+5±1.86E+4 6.65E-2±1.46E-3 129.78±7.53 2

Example 19: Cytokine Release Measurements in vitro in the case of bispecific antibodies against CD3-GUCY2c

T84 cells were treated with T cells (E:T ratio 5:1) and GUCY2C-1608 at different doses. Supernatants were collected at different time points (24 h, 48 h, 96 h), analyzed using a Lumninex multiplex assay according to the manufacturer's instructions, and read on a Luminex 200 using xPONENT software. The cytokines measured were IFN-gamma, IL-10, IL-2, IL-4, IL-6, human TNF-alpha. Figures 5A-5F illustrate the cytokine release profiles of GUCY2C-1608. These cytokines were up-regulated when T84 cells were treated with GUCY2C-1608 in the presence of T cells, confirming the mechanism of T cell-mediated activity of the anti-CD3-GUCY2c bispecific antibody in GUCY2c-expressing cells.

Example 20: Evaluation in vivo activity mediated by bispecific antibody against CD3-GUCY2c

Human PBMCs were thawed in media (X-VIVO 15 (Lonza). 5% human serum albumin (Gemini #100-318), 1% penicillin/streptomycin, 0.01 mM 2-mercaptoethanol was added to approximately 5 million cells per ml. Cells centrifuged and resuspended in Robosep buffer (Stem Cell Technologies) at a concentration of 50 million cells per ml T cells were isolated using the EasySep human T cell enrichment kit (Stem Cell Technologies) T cells were activated and grown using the activation kit /human T cell expansion (Miltenyi) On day 2, T cells were transferred to a G-Rex cell culture apparatus for expansion, media were supplemented with IL-2 (Stem Cell Technologies) and replenished 2 days later. 1 week after activation/expansion At the time of collection, particles were removed using a magnet and cells were resuspended in DPBS at 1 x 10 ⁷ cells/ml for in vivo inoculation.

For xenograft studies, NSG mice were inoculated subcutaneously in the flank with colorectal tumor cell lines (T84, H55, LS1034) or patient-derived xenograft fragments (PDX-CRX-11201, PDX-CRX-12213, PDX-CRX-24225).

Tumors were measured using a digital caliper and volumes were calculated using a modified 1/2 × length × width 2 ellipsoid formula. Mice were randomized and staged according to tumor size of 150–200 mm ³ . An initial dose of anti-GUCY2c bispecific antibody, negative control bispecific antibody, or vehicle was administered to animals on day 0, and 2 million cultured T cells were inoculated the next day. Mice were dosed as bolus injections of 0.2 ml weekly up to 5 times, and all compound and T cell administrations were intravenous through the lateral tail vein of each animal. Tumors were measured twice weekly along with continuous monitoring for signs of graft-versus-host disease (Figures 6-15). All CD3-GUCY2c bispecific antibodies demonstrated dose-dependent T cell-mediated antitumor activity in xenograft cell line and patient-derived xenograft models.

Example 21: Cyclic Guanosine Monophosphate (cGMP) Assay to Characterize Anti-GUCY2c-CD3 Bispecific Antibodies for Potential GUCY2c Pathway Modulation Activity

To test cyclic guanosine monophosphate (cGMP) production by T84 cells in response to GUCY2c pathway stimulation, 2 x 10 ⁶ T84 cells were seeded in a 24-well plate overnight. Cells were washed with DMEM containing 50 mM HEPES and treated with DMEM containing 50 mM HEPES and 1 mM 3-isobutyl-1-methylxanthine (IBMX) for 15 minutes. Cells were then treated with GUCY2C-1608 or a negative control bispecific antibody at concentrations ranging from 0.1 to 99 μM. As a positive control, the GUCY2c agonist STp enterotoxin (BACHEM) was added (0.02 to 51 μM) for 45 min. Particulate matter was removed by centrifugation, and 200 μl of supernatant was used to assay cGMP production using the cGMP Parameter Assay Kit (R&D Systems) according to the manufacturer's instructions. To assess the potential neutralizing activity of GUCY2C-1608, 0.2 μM STp was added in the same range as GUCY2C-1608 or the negative control bispecific antibody as described above for 45 min, and the supernatants were similarly analyzed for cGMP levels.

GUCY2c pathway activity was assessed in T84 cells by measuring cGMP production. Although the GUCY2c pathway agonist bacterial enterotoxin STp increased cGMP in a dose-dependent manner, cGMP production was not increased by the addition of increasing concentrations of GUCY2C-1608. In addition, GUCY2C-1608 had no effect on cGMP production induced by 0.2 μM STp (Figure 16). These results indicate that GUCY2C-1608 is not an agonist of the GUCY2c pathway and does not neutralize GUCY2c pathway function in the presence of ligand.

Example 22: Polyspecificity ELISA for DNA and Insulin

Nonspecific binding of antibodies to molecules other than the target has been suggested to be a mechanism for rapid clearance in vivo (Hötzel et al., 2010, MAbs 4(6):753-760). Evidence for such polyreactive nonspecific binding can be obtained by measuring the binding of membrane preparations (Xu et al., Protein Eng. Des. Select. 26(10):663-70, 2013), baculovirus particles (Hötzel et al., mAbs 4, 753- 760, (2012)) or negatively charged substrates such as DNA, insulin and heparin (Tiller et al., J. Immunol. Methods 329(1-2):112-124, 2008). The DNA and insulin ELISA used a low stringency method originally developed for the detection of low affinity autoantibodies from patients with systemic lupus erythematosus (Tiller et al., J. Immunol. Methods 329, 112, 2008; US Patent No. 7314622).

384-well ELISA plates (Nunc Maxisorp) were coated overnight at 4°C with DNA (10 μg/ml) and insulin (5 μg/ml) in PBS, pH 7.5. ELISA adapted from assays described in Tiller et al., J. Immunol. Methods 329, 112, 2008; US patent No. 7314622, was carried out using a PerkinElmer Janus automated liquid handling device. Wells were washed with water, blocked with 50 μl of polyreactivity ELISA buffer (PEB; PBS containing 0.5% Tween-20, 1 mM EDTA) for 1 hour at room temperature and washed three times with water. Serially diluted mAbs in 25 μl were added to the wells in quadruplicate and incubated for 1 hour at room temperature. The plates were washed 3 times with water and 25 μl of 10 ng/ml goat anti-human IgG antibody (Fcγ-fragment-specific) conjugated to horseradish peroxidase (Jackson ImmunoResearch) was added to each well. The plates were incubated for 1 hour at room temperature, washed 3 times with 80 μl of water, and 25 μl of TMB substrate (Sigma Aldrich) was added to each well. Reactions were stopped after 6 minutes 50 seconds by adding 25 μl of 0.18 M orthophosphoric acid to each well and absorbance was read at 450 nm. DNA and insulin binding scores were calculated as the ratio of the ELISA antibody signal at 10 μg/ml to the signal in the well containing the buffer. Table 34 shows the polyspecificity analysis of anti-CD3-GUCY2c bispecific antibodies using DNA-insulin binding ELISA. The optimized bispecific antibodies showed low nonspecific binding potential.

Table 34

Clone DNA polyreactivity scores Insulin polyreactivity scores GUCY2C-0405 3 1 GUCY2C-0486 2 1 GUCY2C-1186 13 7 GUCY2C-1467 6 7 GUCY2C-1476 7 7 GUCY2C-1478 5 7 GUCY2C-1481 4 5 GUCY2C-1518 6 3 GUCY2C-1526 6 4 GUCY2C-1538 13 7 GUCY2C-1554 3 2 GUCY2C-1555 4 2 GUCY2C-1556 3 1 GUCY2C-1557 5 3 GUCY2C-1590 2 1 GUCY2C-1591 3 2 GUCY2C-1592 4 2 GUCY2C-1606 2 1 GUCY2C-1608 3 2

Example 23: Affinity Capture Nanoparticle Self-Interaction Spectroscopic Analysis (AC-SINS)

Antibody and antibody-like proteins have the potential to interact with themselves, especially at elevated concentrations. This self-interaction can lead to formulation-related viscosity issues during drug development, as well as an increased risk of clearance (Avery et al., mAbs, 2018, Vol. 0, No. 0, 1-12). Self-interaction spectroscopic analysis of affinity capture nanoparticles measures self-interaction and is used to help predict high viscosity and potentially poor pharmacokinetic properties.

The AC-SINS assay was standardized in a 384-well format using a Perkin-Elmer Janus automated liquid handler. 20 nm gold nanoparticles (Ted Pella, Inc., #15705) were coated with a mixture of 80% goat anti-human Fc antibody (Jackson ImmunoResearch Laboratories, Inc. #109-005-098) and 20% nonspecific goat polyclonal antibody (Jackson ImmunoResearch Laboratories, Inc. . # 005-000-003), buffer exchanged to 20 mM sodium acetate, pH 4.3, and diluted to 0.4 mg/ml. After incubation for one hour at room temperature, unoccupied sites on gold nanoparticles were blocked with thiolated polyethylene glycol (2 kDa). The coated nanoparticles were then concentrated 10-fold using a syringe filter and 10 μL was added to 100 μL of mAb at 0.05 mg/mL in PBS, pH 7.2. The coated nanoparticles were incubated with the antibody of interest for 2 h in a 96-well polypropylene plate and then transferred to a 384-well polystyrene plate and read using a Tecan M1000 spectrophotometer. Absorbance was read at 450–650 in 2 nm increments, and Microsoft Excel macro was used to identify maximum absorbance, smooth the data, and fit the data using a second order polynomial. The smoothed maximum absorbance of the average blank sample (PBS buffer only) was subtracted from the smoothed maximum absorbance of the antibody sample to determine the AC-SINS scores of the antibody. Table 35 shows the AC-SINS nonspecificity analysis for the optimized anti-CD3-GUCY2c bispecific antibodies, which showed low potential for self-interaction and nonspecificity.

Table 35

Clone AC-SINS GUCY2C-0405 4 GUCY2C-0486 7 GUCY2C-1186 2 GUCY2C-1467 2 GUCY2C-1476 2 GUCY2C-1478 2 GUCY2C-1481 2 GUCY2C-1518 1 GUCY2C-1526 2 GUCY2C-1538 2 GUCY2C-1554 1 GUCY2C-1555 1 GUCY2C-1556 0 GUCY2C-1557 1 GUCY2C-1590 0 GUCY2C-1591 1 GUCY2C-1592 1 GUCY2C-1608 1

Example 24: Stable Cell Line Generation, Expression and Purification of GUCY2c-1608 Bispecific Antibody

The two chains encoding the bispecific antibody GUCY2C-1608 were cloned into the mammalian expression vector pRY19-GA-Q containing a dual promoter and a recombination site for single site integration (SSI) (Zhang, L. et al. Biotechnology Progress., 31: 1645 -1656, 2015) into the genome of the CHO cell. SSI is based on the insertion of a gene of interest using a recombinase-based cassette replacement at a chromosomal region known for high stable gene expression. The GUCY2C-1608-hollow and GUCY2C-1608-overhang constructs were cloned into the expression vector in cassette one and two, respectively. Using the resulting plasmid GUCY2C-1608, 10E9 CHO-K1 SV cells were transfected by electroporation using the pRY19 vector containing the gene of interest, followed by positive and negative selection pressure using hygromycin-B and ganciclovir. These CHO populations were subjected to a 3-week recovery phase. When the observed viability exceeded 90%, cell banks were obtained for subsequent use. The resulting CHO populations were then subjected to 12 days of expression on a fed-batch culture platform in CD-CHO media (ThermoFisher Scientific, Waltham, MA). At the end of the 12-day period, cells were centrifuged at 3,000 × g for 10 minutes, followed by sterile filtration of conditioned media using a 0.2-μm PES bottle plug filter (Corning, Oneonta, NY). In addition, stable CHO populations were scaled to a 10 L controlled swing bioreactor (Finesse, Santa Clara, CA). CHO populations were grown in Pfizer's proprietary media in a 12-day fed-batch culture that included chemically defined feed to maintain culture viability. Once complete, the culture was subjected to depth filtration using a filtration path consisting of a 20" 5 µm Pall Profile II filter followed by a 10" 0.2 µm Pall Supor filter (Pall Corporation, Port Washington, NY). Titers were assessed using Protein A affinity chromatography. Protein purification was then carried out by methods known in the art, including Protein A chromatography, hydrophobic interaction chromatography and ion exchange chromatography. The expression titer of GUCY2C-1608 protein from the stably transfected CHO cell population is shown in Table 36. Expression titers for the GUCY2C-1608 clone populations indicated expression levels ranging from 0.48 to 0.84 g/liter.

Table 36

Clone population Expression (g/l) Population A 0.842 Population B 0.710 Population C 0.480

Example 25: Mass Spectrometry Analysis of Bispecific Antibody GUCY2C-1608

LC/MS analysis was performed to confirm the formation of the correct paired purified bispecific antibody GUCY2C-1608. The molecular weights of a bispecific molecule are determined from its unique amino acid sequences, and by determining the exact molecular weight, evidence of the presence of correctly paired anti-GUCY2c bispecific molecules is obtained. Briefly, a sample of the anti-CD3-GUCY2c bispecific antibody was analyzed by LC/MS analysis using a Waters Acquity H-class HPLC coupled to a Bruker maXis II Q-ToF mass spectrometer. Analytes were loaded onto an SMT SAM C2 column (2.1 mm × 100 mm, maintained at 80°C) and eluted using a tertiary gradient of buffer B (80% 1-propanol, 20% acetonitrile with 0.05% trifluoroacetic acid) and buffer C (100% acetonitrile with 0.05% trifluoroacetic acid) at a flow rate of 300 µl/min (5 min ramp to 19% B and 4% C, 20 min ramp to 22% B and 16% C, 10 min ramp increasing to 0% B and 98% C). Mass spectrometric detection was carried out in positive mode with a capillary voltage of 3.5 kV. Data analysis was performed using MaxEnt 1 deconvolution in Compass DataAnalysis v4.2. Figure 17 shows the high purity at predicted molecular weight of the final purified GUCY2C-1608 bispecific antibody.

Example 26: Sequences

Tables 37A and 37B summarize the SEQ ID NOs of the antibodies or antibody fragments provided herein. Table 37A uses the ABM numbering system for VH CDR1 sequences. Table 37A uses the Kabat numbering system for all other sequences, including CDR2 VH.

Table 37A

Subsequence CDR_H1 CDR_H2 CDR_H3 CDR_L1 CDR_L2 CDR_L3 FV_H FV_L FW_H1 FW_H2 FW_H3 FW_H4 FW_L1 FW_L2 FW_L3 FW_L4 Linker 1 Linker 2 CD3-0001 2 3 4 77 78 79 1 76 5 6 7 8 80 81 82 83 N.A. N.A. CD3-0004 2 3 4 85 78 79 1 84 5 6 7 8 86 87 88 89 But But CD3-0006 2 10 4 91 78 79 9 90 5 6 7 8 80 81 82 83 But But GUCY2C-0074 12 13 14 93 94 95 eleven 92 15 16 17 18 96 97 98 99 190 But GUCY2C-0077 20 21 22 101 102 95 19 100 23 24 25 18 96 97 103 99 190 But GUCY2C-0078 20 21 22 105 102 95 19 104 23 24 25 18 96 97 103 99 190 But GUCY2C-0098 27 28 29 107 108 109 26 106 thirty 16 31 32 96 110 111 99 190 But GUCY2C-0104 34 35 36 113 114 115 33 112 37 38 39 40 116 110 117 118 190 But GUCY2C-0105 42 43 44 120 78 121 41 119 45 46 47 32 122 123 124 99 190 But GUCY2C-0240 12 13 14 93 94 95 48 125 5 49 50 51 126 127 82 128 190 But GUCY2C-0315 12 53 14 93 94 95 52 129 54 55 56 51 130 131 132 133 But But GUCY2C-0179 42 43 44 120 78 121 57 134 5 49 58 59 80 135 82 83 But But GUCY2C-0193 27 28 29 107 108 109 60 136 5 49 61 51 80 81 82 128 But But GUCY2C-0210 27 28 29 107 108 109 62 137 5 63 7 51 126 81 82 128 But But GUCY2C-0212 27 28 29 107 108 109 64 137 5 49 7 51 126 81 82 128 But But GUCY2C-0241 27 28 29 107 108 109 60 137 5 49 61 51 126 81 82 128 But But GUCY2C-0247 27 28 29 107 108 109 60 138 5 49 61 51 126 139 82 128 190 But GUCY2C-1186 27 66 29 107 141 142 65 140 5 49 61 51 126 81 82 128 190 But GUCY2C-1467 27 28 29 107 144 109 60 143 5 49 61 51 126 81 82 128 190 But GUCY2C-1476 27 68 29 107 146 142 67 145 5 49 61 51 126 81 82 128 190 But GUCY2C-1478 27 70 29 148 149 142 69 147 5 49 61 51 126 81 82 128 190 191 GUCY2C-1481 27 70 29 107 151 142 69 150 5 49 61 51 126 81 82 128 190 But GUCY2C-1512 27 72 29 153 141 154 71 152 5 49 61 51 126 81 82 155 190 But GUCY2C-1518 27 70 29 153 157 142 69 156 5 49 61 51 126 81 82 128 190 But GUCY2C-1526 27 68 29 153 159 142 67 158 5 49 61 51 126 81 82 128 190 But GUCY2C-1527 27 70 29 107 161 154 69 160 5 49 61 51 126 81 82 128 190 But GUCY2C-1538 27 68 29 163 164 165 67 162 5 49 61 51 126 81 82 128 190 But GUCY2C-1554 74 75 29 167 168 169 73 166 5 49 61 51 80 81 82 128 190 But GUCY2C-1555 74 75 29 167 149 169 73 170 5 49 61 51 80 81 82 128 190 But GUCY2C-1556 74 75 29 167 168 142 73 171 5 49 61 51 80 81 82 128 190 But GUCY2C-1557 74 75 29 167 149 142 73 172 5 49 61 51 80 81 82 128 190 But GUCY2C-1590 74 75 29 148 168 169 73 173 5 49 61 51 126 81 82 128 190 But GUCY2C-1591 74 75 29 148 149 169 73 174 5 49 61 51 126 81 82 128 190 But GUCY2C-1592 74 75 29 148 168 142 73 175 5 49 61 51 126 81 82 128 190 But GUCY2C-1602 74 75 29 148 149 142 73 147 5 49 61 51 126 81 82 128 190 191 huIGHV3-7 177 178 179 But But But 176 But 5 6 7 But But But But But But But huIGKV1-39 But But But 181 182 183 But 180 But But But But 80 81 82 But But But GUCY2C-0405 27 70 29 148 149 142 69 147 5 49 61 51 126 81 82 128 But But GUCY2C-0486 27 70 29 148 149 142 69 147 5 49 61 51 126 81 82 128 But But GUCY2C-1640 74 75 29 148 149 142 73 147 5 49 61 51 126 81 82 128 But But GUCY2C-0250 27 28 29 107 108 109 60 137 5 49 61 51 126 81 82 128 190 But

Table 37B

Subsequence Linker 3 Linker 4 Linker 5 Linker 6 CL1 human IgG1 CH1 human IgG1 Human IgG1 hinge region Human IgG1 Fc (CH2-CH3) CH vs CD3 VL vs CD3 Fc chain with lug Fc chain with cavity IgG light chain IgG heavy chain VL with projection VH with cavity Full sequence Nucleotides of the first polypeptide chain Nucleotides of the second polypeptide chain CD3-0001 But But But But But But But But 1 76 But But But But But But But But But CD3-0004 But But But But But But But But 1 84 But But But But But But But But But CD3-0006 But But But But But But But But 9 90 But But But But But But But But But GUCY2C-0074 192 But But But But But But But 1 84 188 189 But But 196 197 198 But But GUCY2C-0077 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-0078 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-0098 192 But But But But But But But 1 84 188 189 But But 199 200 201 But But GUCY2C-0104 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-0105 192 But But But But But But But 1 84 188 189 But But 202 203 204 But But GUCY2C-0240 192 But But But But But But But 1 84 188 189 But But 205 206 207 But But GUCY2C-0315 But But But But 184 185 186 187 But But But But 214 223 But But But But But GUCY2C-0179 But But But But 184 185 186 187 But But But But 214 223 But But But But But GUCY2C-0193 But But But But 184 185 186 187 But But But But 214 223 But But But But But GUCY2C-0210 But But But But 184 185 186 187 But But But But 214 223 But But But But But GUCY2C-0212 But But But But 184 185 186 187 But But But But 214 223 But But But But But GUCY2C-0241 But But But But 184 185 186 187 But But But But 208 209 But But But But But GUCY2C-0247 192 But But But But But But But 1 84 188 189 But But 210 211 212 But But GUCY2C-1186 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1467 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1476 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1478 192 But But But But But But But 9 90 188 189 But But 216 217 218 But But GUCY2C-1481 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1512 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1518 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1526 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1527 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1538 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1554 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1555 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1556 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1557 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1590 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1591 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1592 192 But But But But But But But 1 84 188 189 But But But But But But But GUCY2C-1608 192 But But But But But But But 9 90 188 189 But But 216 220 221 246 247 hulGHV3-7 But But But But But But But But But But But But But But But But But But But hulGKV1-39 But But But But But But But But But But But But But But But But But But But GUCY2C-0405 But 193 194 195 But But But 187 But But But But But But But But 213 But But GUCY2C-0486 But But But But 184 185 186 187 But But But But 214 215 But But But But But GUCY2C-1640 But But But But 184 185 186 187 But But But But 214 223 But But But But But GUCY2C-0250 But But But But But But But But 1 76 188 189 But But 248 249 250 But But

Table 38 shows the sequences presented herein.

Table 38

SEQ ID NO: Description Subsequence 1 CD3-0001_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSS 2 CD3-0001_VH,
2B5-1038 VH,
2B5-1039 VH,
2B5-1040 VH,
CDR1 (extended/ABM) GFTFSDYYMT 3 CD3-0001_CDR2 VH FIRNRARGYTSDHNPSVKG 4 CD3-0001_VH,
2B5-1038 VH,
2B5-1039 VH,
2B5-1040 VH,
CDR3 DRPSYYVLDY 5 CD3-0001_VH FW_H1 EVQLVESGGGLVQPGGSLRLSCAAS 6 CD3-0001_VH FW_H2 WVRQAPGKGLEWVA 7 CD3-0001_VH FW_H3 RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR 8 CD3-0001_VH FW_H4 WGQGTTVTVSS 9 CD3-0006_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSS 10 CD3-0006_VH,
2B5-1038 VH,
2B5-1039 VH,
2B5-1040 VH,
CDR2 (Kabat) FIRNQARGYTSDHNPSVKG eleven GUCY2C-0074_VH EVQLQQSGAELARPGASVNLSCKASGYTFTTYWMQWVKQRPGQGLEWIGAIYPGDGMTTYTQKFKDKATLTADKSSSTAYMQLSSLASEDSAVYYCVRKGMDYWGQGTSVTVSS 12 GUCY2C-0074_CDR1 VH GYTFTTYWMQ 13 GUCY2C-0074_CDR2 VH AIYPGDGMTTYTQKFKD 14 GUCY2C-0074_CDR3 VH KGMDY 15 GUCY2C-0074_VH FW_H1 EVQLQQSGAELARPGASVNLSCKAS 16 GUCY2C-0074_VH FW_H2 WVKQRPGQGLEWIG 17 GUCY2C-0074_VH FW_H3 KATLTADKSSSTAYMQLSSLASEDSAVYYCVR 18 GUCY2C-0074_VH FW_H4 WGQGTSVTVSS 19 GUCY2C-0077_VH EVQLQQSGAELARPGASVKLSCKASGYTFTKYWMQWIKQRPGQGLEWIGAIYPGDGFTTYTQKFKGKATLTADKSSNTAYMQLSSLASEDSAVYYCARRNYGRTYGGDYWGQGTSVTVSS 20 GUCY2C-0077_CDR1 VH GYTFTKYWMQ 21 GUCY2C-0077_CDR2 VH AIYPGDGFTTYTQKFKG 22 GUCY2C-0077_CDR3 VH RNYGRTYGGDY 23 GUCY2C-0077_VH FW_H1 EVQLQQSGAELARPGASVKLSCKAS 24 GUCY2C-0077_VH FW_H2 WIKQRPGQGLEWIG 25 GUCY2C-0077_VH FW_H3 KATLTADKSSNTAYMQLSSLASEDSAVYYCAR 26 GUCY2C-0098_VH QVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTVDKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSS 27 GUCY2C-0098_CDR1 VH GYTFTSYWMH 28 GUCY2C-0098_CDR2 VH EIKPSNGLTNYIEKFKN 29 GUCY2C-0098_CDR3 VH TITTTEGYWFFDV thirty GUCY2C-0098_VH FW_H1 QVQLQQPGAELVKPGASVKLSCKAS 31 GUCY2C-0098_VH FW_H3 KATLTVDKSATTAYMQLSSLTAEDSAVYYCTR 32 GUCY2C-0098_VH FW_H4 WGAGTTVTVSS 33 GUCY2C-0104_VH EVQLQQSGAELVKPGASVKLSCTASGFNIKDTYIHWVKQRPEQGLEWIGRIDPANGNANYDPKFQGKATITADTSSNTAYLQLSSLTSEDTAVFYCSSLGTGTYWGQGTTLTVSS 34 GUCY2C-0104_CDR1 VH GFNIKDTYIH 35 GUCY2C-0104_CDR2 VH RIDPANGNANYDPKFQG 36 GUCY2C-0104_CDR3 VH LGTGTY 37 GUCY2C-0104_VH FW_H1 EVQLQQSGAELVKPGASVKLSCTAS 38 GUCY2C-0104_VH FW_H2 WVKQRPEQGLEWIG 39 GUCY2C-0104_VH FW_H3 KATITADTSSNTAYLQLSSLTSEDTAVFYCSS 40 GUCY2C-0104_VH FW_H4 WGQGTTLTVSS 41 GUCY2C-0105_VH EVQLQQSGPELVKPGASVKISCKASGYSFTDYIMLWVKQSHGKSLEWIGNSNPYYGSTSYNLKFKGKATLTVDKSSSTAYMHLNSLTSEDSAVYYCARSGYYGSSPYWYFDVWGAGTTVTVSS 42 GUCY2C-0105_CDR1 VH GYSFTDYIML 43 GUCY2C-0105_CDR2 VH NSNPYYGSTSYNLKFKG 44 GUCY2C-0105_CDR3 VH SGYYGSSPYWYFDV 45 GUCY2C-0105_VH FW_H1 EVQLQQSGPELVKPGASVKISCKAS 46 GUCY2C-0105_VH FW_H2 WVKQSHGKSLEWIG 47 GUCY2C-0105_VH FW_H3 KATLTVDKSSSTAYMHLNSLTSEDSAVYYCAR 48 GUCY2C-0240_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTTYWMQWVRQAPGKGLEWIGAIYPGDGMTTYTQKFKDRFTISADKAKNSAYLQMNSLRAEDTAVYYCVRKGMDYWGQGTLVTVSS 49 GUCY2C-0240_VH FW_H2 WVRQAPGKGLEWIG 50 GUCY2C-0240_VH FW_H3 RFTISADKAKNSAYLQMNSLRAEDTAVYYCVR 51 GUCY2C-0240_VH FW_H4 WGQGTLVTVSS 52 GUCY2C-0315_VH QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMQWVRQAPGQGLEWIGAIYPGDGMTTYTQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCVRKGMDYWGQGTLVTVSS 53 GUCY2C-0315_CDR2 VH AIYPGDGMTTYTQKFQG 54 GUCY2C-0315_VH FW_H1 QVQLVQSGAEVKKPGASVKVSCKAS 55 GUCY2C-0315_VH FW_H2 WVRQAPGQGLEWIG 56 GUCY2C-0315_VH FW_H3 RVTMTRDTSTSTVYMELSSLRSEDTAVYYCVR 57 GUCY2C-0179_VH EVQLVESGGGLVQPGGSLRLSCAASGYSFTDYIMLWVRQAPGKGLEWIGNSNPYYGSTSYNLKFKGRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARSGYYGSSPYWYFDVWGQGTMVTVSS 58 GUCY2C-0179_VH FW_H3 RFTISVDKAKNSAYLQMNSLRAEDTAVYYCAR 59 GUCY2C-0179_VH FW_H4 WGQGTMVTVSS 60 GUCY2C-0193_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 61 GUCY2C-0193_VH FW_H3 RFTISVDKAKNSAYLQMNSLRAEDTAVYYCTR 62 GUCY2C-0210_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIAEIKPSNGLTNYIEKFKNRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARTITTTEGYWFFDVWGQGTLVTVSS 63 GUCY2C-0210_VH FW_H2 WVRQAPGKGLEWIA 64 GUCY2C-0212_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARTITTTEGYWFFDVWGQGTLVTVSS 65 GUCY2C-1186_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNIHPKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 66 GUCY2C-1186_CDR2 VH EIKPSNGLTNIHPKFKN 67 GUCY2C-1476_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYNEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 68 GUCY2C-1476_CDR2 VH EIKPSNGLTNYNEKFKN 69 GUCY2C-1478_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNVHEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 70 GUCY2C-1478_CDR2 VH EIKPSNGLTNVHEKFKN 71 GUCY2C-1512_VH EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYAEQFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 72 GUCY2C-1512_CDR2 VH EIKPSNGLTNYAEQFKN 73 GUCY2C-1554_VH;
GUCY2C-1608_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 74 GUCY2C-1554_CDR1 VH GFTFSSYWMH 75 GUCY2C-1554_CDR2 VH EIKPSNELTNVHEKFKD 76 CD3-0001_VL DIQMTQSPSSSLSASVGDRVTITCTSSQSLFNVRSRKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIK 77 CD3-0001_CDR1 VL TSSQSLFNVRSRKNYLA 78 CD3-0001_CDR2 VL WASTRES 79 CD3-0001_CDR3 VL KQSYDLFT 80 CD3-0001_VL FW_L1 DIQMTQSPSSLSSASVGDRVTITTC 81 CD3-0001_VL FW_L2 WYQQKPGKAPKLLIY 82 CD3-0001_VL FW_L3 GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC 83 CD3-0001_VL FW_L4 FGGGTKVEIK 84 CD3-0004_VL DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIK 85 CD3-0004_CDR1 VL KSSQSLFNVRSRKNYLA 86 CD3-0004_VL FW_L1 DIVMTQSPDSLAVSLGERATINC 87 CD3-0004_VL FW_L2 WYQQKPGQPPKLLIS 88 CD3-0004_VL FW_L3 GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC 89 CD3-0004_VL FW_L4 FGSGTKLEIK 90 CD3-0006_VL DIQMTQSPSSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIK 91 CD3-0006_CDR1 VL TSSQSLFNVRSQKNYLA 92 GUCY2C-0074_VL DIVLTQSPASLAVSLGQRATISCRASESVDNYGISFMNWFQQKPGQPPKLLIYAASNPGSGVPARFSGSGSGTDFSLNIHPLEEDDTAMFFCQQSKEVPYTFGGGTKLEIK 93 GUCY2C-0074_CDR1 VL RASESVDNYGISFMN 94 GUCY2C-0074_CDR2 VL AASNPGS 95 GUCY2C-0074_CDR3 VL QQSKEVPYT 96 GUCY2C-0074_VL FW_L1 DIVLTQSPASLAVSLGQRATISC 97 GUCY2C-0074_VL FW_L2 WFQQKPGQPPKLLIY 98 GUCY2C-0074_VL FW_L3 GVPARFSGSGSGTDFSLNIHPLEEDDTAMFFC 99 GUCY2C-0074_VL FW_L4 FGGGTKLEIK 100 GUCY2C-0077_VL DIVLTQSPASLAVSLGQRATISCRASESVDNFDISFMNWFQQKPGQPPKLLIYAASNQGSGVPARFSGSGSGTDFSLNIHPMEEDDTAMYFCQQSKEVPYTFGGGTKLEIK 101 GUCY2C-0077_CDR1 VL RASESVDNFDISFMN 102 GUCY2C-0077_CDR2 VL AASNQGS 103 GUCY2C-0077_VL FW_L3 GVPARFSGSGSGTDFSLNIHPMEEDDTAMYFC 104 GUCY2C-0078_VL DIVLTQSPASLAVSLGQRATISCRAGESVDNFDISFMNWFQQKPGQPPKLLIYAASNQGSGVPARFSGSGSGTDFSLNIHPMEEDDTAMYFCQQSKEVPYTFGGGTKLEIK 105 GUCY2C-0078_CDR1 VL RAGESVDNFDISFMN 106 GUCY2C-0098_VL DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIK 107 GUCY2C-0098_CDR1 VL RASESVDYYGTSLMQ 108 GUCY2C-0098_CDR2 VL AASNVES 109 GUCY2C-0098_CDR3 VL QQTRKVYT 110 GUCY2C-0098_VL FW_L2 WYQQKPGQPPKLLIY 111 GUCY2C-0098_VL FW_L3 GVPARFSGSGSGTDFSLNIHPVEEDDIAMYFC 112 GUCY2C-0104_VL DIVMTQSPASLAVSLGQRATISCRASKGVTTSGYSYMHWYQQKPGQPPKLLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSREFPLTFGAGTKLELK 113 GUCY2C-0104_CDR1 VL RASKGVTTSGYSYMH 114 GUCY2C-0104_CDR2 VL LASNLES 115 GUCY2C-0104_CDR3 VL QHSREFPLT 116 GUCY2C-0104_VL FW_L1 DIVMTQSPASLAVSLGQRATISC 117 GUCY2C-0104_VL FW_L3 GVPARFSGSGSGTDFTLNIHPVEEEDAATYYC 118 GUCY2C-0104_VL FW_L4 FGAGTKLELK 119 GUCY2C-0105_VL DIVMTQSPSSLAVSVGEKVTVSCKSSQSLLYSSNQKNYLAWYQQRPGQSPKLLIYWASTRESGVPDRFTGSGSGTDFTLSISSVKAEDLAVYYCQQYYSYPTFGGGTKLEIK 120 GUCY2C-0105_CDR1 VL KSSQSLLYSSNQKNYLA 121 GUCY2C-0105_CDR3 VL QQYYSYPT 122 GUCY2C-0105_VL FW_L1 DIVMTQSPSSLAVSVGEKVTVSC 123 GUCY2C-0105_VL FW_L2 WYQQRPGQSPKLLIY 124 GUCY2C-0105_VL FW_L3 GVPDRFTGSGSGTDFTLSISSVKAEDLAVYYC 125 GUCY2C-0240_VL DIQLTQSPSSLSASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNPGSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSKEVPYTFGQGTKLEIK 126 GUCY2C-0240_VL FW_L1 DIQLTQSPSSLSSASVGDRVTITTC 127 GUCY2C-0240_VL FW_L2 WFQQKPGKAPKLLIY 128 GUCY2C-0240_VL FW_L4 FGQGTKLEIK 129 GUCY2C-0315_VL EIVLTQSPATLSLSPGERATLSCRASESVDNYGISFMNWYQQKPGQAPRLLIYAASNPGSGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSKEVPYTFGQGTKVEIK 130 GUCY2C-0315_VL FW_L1 EIVLTQSPATLSLSPGERATLSC 131 GUCY2C-0315_VL FW_L2 WYQQKPGQAPRLLIY 132 GUCY2C-0315_VL FW_L3 GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC 133 GUCY2C-0315_VL FW_L4 FGQGTKVEIK 134 GUCY2C-0179_VL DIQMTQSPSSSLSASVGDRVTITCKSSQSLLYSSNQKNYLAWYQQKPGKSPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYSYPTFGGGTKVEIK 135 GUCY2C-0179_VL FW_L2 WYQQKPGKSPKLLIY 136 GUCY2C-0193_VL DIQMTQSPSSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIK 137 GUCY2C-0210_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIK 138 GUCY2C-0247_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKPPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIK 139 GUCY2C-0247_VL FW_L2 WYQQKPGKPPKLLIY 140 GUCY2C-1186_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASKRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 141 GUCY2C-1186_CDR2 VL AASKRYS 142 GUCY2C-1186_CDR3 VL QQTRKAYT 143 GUCY2C-1467_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASKLWSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIK 144 GUCY2C-1467_CDR2 VL AASKLWS 145 GUCY2C-1476_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASKVAPGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 146 GUCY2C-1476_CDR2 VL AASKVAP 147 GUCY2C-1478_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 148 GUCY2C-1478_CDR1 VL RASESVDYYGSSLLQ 149 GUCY2C-1478_CDR2 VL AASKLAS 150 GUCY2C-1481_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNIAPGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 151 GUCY2C-1481_CDR2 VL AASNIAP 152 GUCY2C-1512_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKEYTFGQGTKLKIK 153 GUCY2C-1512_CDR1 VL RASESVDYYGSSLMQ 154 GUCY2C-1512_CDR3 VL QQTRKEYT 155 GUCY2C-1512_VL FW_L4 FGQGTKLKIK 156 GUCY2C-1518_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASNVAPGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 157 GUCY2C-1518_CDR2 VL AASNVAP 158 GUCY2C-1526_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASHRASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 159 GUCY2C-1526_CDR2 VL AASHRAS 160 GUCY2C-1527_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKEYTFGQGTKLEIK 161 GUCY2C-1527_CDR2 VL AASNVAS 162 GUCY2C-1538_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGHSLMQWYQQKPGKAPKLLIYAASNRYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYSFGQGTKLEIK 163 GUCY2C-1538_CDR1 VL RASESVDYYGHSLMQ 164 GUCY2C-1538_CDR2 VL AASNRYS 165 GUCY2C-1538_CDR3 VL QQTRKAYS 166 GUCY2C-1554_VL DIQMTQSPSSLSASVGDRVTITCRASQSVDYYGSSLLQWYQQKPGKAPKLLIYDASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQERKAYTFGQGTKLEIK 167 GUCY2C-1554_CDR1 VL RASQSVDYYGSSLLQ 168 GUCY2C-1554_CDR2 VL DASKLAS 169 GUCY2C-1554_CDR3 VL QQERKAYT 170 GUCY2C-1555_VL DIQMTQSPSSLSASVGDRVTITCRASQSVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQERKAYTFGQGTKLEIK 171 GUCY2C-1556_VL DIQMTQSPSSLSASVGDRVTITCRASQSVDYYGSSLLQWYQQKPGKAPKLLIYDASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 172 GUCY2C-1557_VL DIQMTQSPSSLSASVGDRVTITCRASQSVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 173 GUCY2C-1590_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYDASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQERKAYTFGQGTKLEIK 174 GUCY2C-1591_VL DIQLTQSPSSLSSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQERKAYTFGQGTKLEIK 175 GUCY2C-1592_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYDASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 176 huIGHV3-7_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARYYYYYGMDV 177 huIGHV3-7_CDR1 VH GFTFSSYWMS 178 huIGHV3-7_CDR2 VH NIKQDGSEKYYVDSVKG 179 huIGHV3-7_CDR3 VH YYYYYGMDV 180 huIGKV1-39_VL DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIK 181 huIGKV1-39_CDR1 VL RASQSISSYLN 182 huIGKV1-39_CDR2 VL AASSLQS 183 huIGKV1-39_CDR3 VL QQSYSTPYT 184 Human IgG1 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 185 CH1 human IgG1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV 186 Human IgG1 hinge region EPKSCDKTHTCPPCP 187 Human Fc IgG1-3m (CH2-CH3) APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK 188 Fc chain with lug APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPG 189 Fc chain with cavity APEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG 190 Linker 1 GGGSGGGG 191 Linker 2 GGGGSGGGG 192 Linker 3 GCPPCP 193 Linker 4 GGGGSGGGGSGGGGSG 194 Linker 5 RTSDQ 195 Linker 6 EPKSSDKTHTCPPCP 196 GUCY2C-0074_protrusion DIVLTQSPASLAVSLGQRATISCRASESVDNYGISFMNWFQQKPGQPPKLLIYAASNPGSGVPARFSGSGSGTDFSLNIHPLEEDDTAMFFCQQSKEVPYTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 197 GUCY2C-0074_cavity DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLQQSGAELARPGASVNLSCKASGYTFTTYWMQWVKQRPGQGLEWIGAIYPGDGMTTYTQKFKDKATLTAD KSSSTAYMQLSSLASEDSAVYYCVRKGMDYWGQGTSVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 198 GUCY2C-0074_monomer DIVLTQSPASLAVSLGQRATISCRASESVDNYGISFMNWFQQKPGQPPKLLIYAASNPGSGVPARFSGSGSGTDFSLNIHPLEEDDTAMFFCQQSKEVPYTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQMN SLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLQQSGAELAR PGASVNLSCKASGYTFTTYWMQWVKQRPGQGLEWIGAIYPGDGMTTYTQKFKDKATLTADKSSSTAYMQLSSLASEDSAVYYCVRKGMDYWGQGTSVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 199 GUCY2C-0098_protrusion DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQ MNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 200 GUCY2C-0098_cavity DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGQVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTV DKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSL SCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 201 GUCY2C-0098_monomer DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYLQ MNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGQVQLQQPGA ELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTVDKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 202 GUCY2C-0105_protrusion DIVMTQSPSSLAVSVGEKVTVSCKSSQSLLYSSNQKNYLAWYQQRPGQSPKLLIYWASTRESGVPDRFTGSGSGTDFTLSISSVKAEDLAVYYCQQYYSYPTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 203 GUCY2C-0105_cavity DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLQQSGPELVKPGASVKISCKASGYSFTDYIMLWVKQSHGKSLEWIGNSNPYYGSTSYNLKFKGKATLTVDK SSSTAYMHLNSLTSEDSAVYYCARSGYYGSSPYWYFDVWGAGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSL SCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 204 GUCY2C-0105_monomer DIVMTQSPSSLAVSVGEKVTVSCKSSQSLLYSSNQKNYLAWYQQRPGQSPKLLIYWASTRESGVPDRFTGSGSGTDFTLSISSVKAEDLAVYYCQQYYSYPTFGGGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLQQ PELVKPGASVKISCKASGYSFTDYIMLWVKQSHGKSLEWIGNSNPYYGSTSYNLKFKGKATLTVDKSSSTAYMHLNSLTSEDSAVYYCARSGYYGSSPYWYFDVWGAGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 205 GUCY2C-0240_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNPGSGVPSRFSGSGSGTDFTISSLQPEDFATYYCQQSKEVPYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYL QMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 206 GUCY2C-0240_cavity DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTTYWMQWVRQAPGKGLEWIGAIYPGDGMTTYTQKFKDRFTISADKAKNS AYLQMNSLRAEDTAVYYCVRKGMDYWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 207 GUCY2C-0240_monomer DIQLTQSPSSLSASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNPGSGVPSRFSGSGSGTDFTISSLQPEDFATYYCQQSKEVPYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLYL QMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLVESGLV QPGGSLRLSCAASGYTFTTYWMQWVRQAPGKGLEWIGAIYPGDGMTTYTQKFKDRFTISADKAKNSAYLQMNSLRAEDTAVYYCVRKGMDYWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 208 GUCY2C-0241_light chain DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 209 GUCY2C-0241_heavy chain EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 210 GUCY2C-0247_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKPPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 211 GUCY2C-0247_cavity DIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNSAY LQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 212 GUCY2C-0247_monomer DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKPPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIVMTQSPDSLAVSLGERATINCKSSQSLFNVRSRKNYLAWYQQKPGQPPKLLISWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCKQSYDLFTFGSGTKLEIKGGGSGGGGEVQLVESG LVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 213 GUCY2C-0405_scFv-Fc EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNVHEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGDIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLA SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKRTSDQEPKSSDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP 214 GUCY2C-0486_light chain; GUCY2C-1640_light chain DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC 215 GUCY2C-0486_heavy chain EVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNVHEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 216 GUCY2C-1478_protrusion;
GUCY2C-1608_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 217 GUCY2C-1478_cavity DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNVHEKFKNRFTISVDKAK NSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 218 GUCY2C-1478_monomer DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVES GGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNVHEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 219 Human IgG1 upper hinge region EPKSCDKTHT 220 GUCY2C-1608_cavity DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 221 GUCY2C-1608_monomer DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVES GGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREESVQYNSTYRVV LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 222 Human IgG1 lower hinge region CPPCP 223 GUCY2C-1640_heavy chain EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 224 Full size human GUCY2C SQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQD KEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYLEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQILMIAVFTLTGA VVLLLLVALLMLRKYRKDYELRQKKWSHIPPENIFPLETNETNHVSLKIDDDKRRDTIQRLRQCKYDKKRVILKDLKHNDGNFTEKQKIELNKLLQIDYYNLTKFYGTVKLDTMIFGVIEYCERGSLREVLNDTISYPDGTFMDWEFKISVLYDIAKGMSYLHSSKTEVHGRLKSTNCVVDSRMVVK ITDFGCNSILPPKKDLWTAPEHLRQANISQKGDVYSYGIIAQEIILRKETFYTLSCRDRNEKIFRVENSNGMKPFRPDLFLETAEEKELEVYLLVKNCWEEDPEKRPDFKKIETTLAKIFGLFHDQKNESYMDTLIRRLQLYSRNLEHLVEERTQLYKAERDRADRLNFMLLPRLVVKSLKEKGFVEPELYEEVTIYFSDIVGF TTICKYSTPMEVVDMLNDIYKSFDHIVDHHDVYKVETIGDAYMVASGLPKRNGNRHAIDIAKMALEILSFMGTFELEHLPGLPIWIRIGVHSGPCAAGVVGIKMPRYCLFGDTVNTASRMESTGLPLRIHVSGSTIAILKRTECQFLYEVRGETYLKGRGNETTYWLTGMKDQKFNLPTPPTVENQQRLQAEFSDMIANSLQKRQ AAGIRSQKPRRVASYKKGTLEYLQLNTTDKESTYFTRTRPLEQKLISEEDLAANDILDYKDDDDK 225 Full size human GUCY2C TCCCAGGTGAGTCAGAACTGCCACAATGGCAGCTATGAAATCAGCGTCCTGATGATGGGCAACTCAGCCTTTGCAGAGCCCCTGAAAAACTTGGAAGATGCGGTGAATGAGGGGCTGGAAATAGTGAGAGGACGTCTGCAAAATGCTGGCCTAAATGTGACTGTGAACGCTACTTTCATGTATTCGGATGGTCTGATTCATAACTCAGGCGACTGCCGGAGTAGCACCTGTGAAGGCCTCGACCTACTCA GGAAAATTTCAAATGCACAACGGATGGGCTGTGTCCTCATAGGGCCCTCATGTACATACTCCACCTTCCAGATGTACCTTGACACAGAATTGAGCTACCCCATGATCTCAGCTGGAAGTTTTGGATTGTCATGTGACTATAAAGAAACCTTAACCAGGCTGATGTCTCCAGCTAGAAAGTTGATGTACTTCTTGGTTAACTTTTGGAAAACCAACGATCTGCCCTTCAAAACTTATTCCTGGAGCACTTCGTATG TTTACAAGAATGGTACAGAAACTGAGGACTGTTTCTGGTACCTTTAATGCTCTGGAGGCTAGCGTTTCCTATTTCTCCCACGAACTCGGCTTTAAGGTGGTGTTAAGACAAGATAAGGAGTTTCAGGATATCTTAATGGACCACAACAGGAAAAGCAATGTGATTATTATGTGTGGTGGTCCAGAGTTCCTCTACAAGCTGAAGGGTGACCGAGCAGTGGCTGAAGACATTGTCATTATTCT AGTGGATCTTTTCAATGACCAGTACTTGGAGGACAATGTCACAGCCCCTGACTATATGAAAAATGTCCTTGTTCTGACGCTGTCTCCTGGGAATTCCCTTCTAAATAGCTCTTTCTCCAGGAATCTATCACCAACAAAACGAGACTTTGCTCTTGCCTATTTGAATGGAATCCTGCTCTTTGGACATATGCTGAAGATATTTCTTGAAAATGGAGAAAATATTACCACCCCCAAATTTGCTCATGCTTTCAGGAATC TCACTTTTGAAGGGTATGACGGTCCAGTGACCTTGGATGACTGGGGGGATGTTGACAGTACCATGGTGCTTCTGTATACCTCTGTGGACACCAAGAAATACAAGGTTCTTTTGACCTATGATACCCACGTAAATAAGACCTATCCTGTGGATATGAGCCCCACATTCACTTGGAAGAACTCTAAACTTCCTAATGATATTACAGGCCGGGGCCCTCAGATCCTGATGATTGCAGGGTCTTCACCTCACT AGCTGTGGTGCTGCTCCTGCTCGTCGCTCTCCTGATGCTCAGAAAATATAGAAAAGATTATGAACTTCGTCAGAAAAAATGGTCCCACATTCCTCCTGAAAATATCTTTCCTCTGGAGACCAATGAGACCAATCATGTTAGCCTCAAGATCGATGATGACAAAAGACGAGATACAATCCAGAGACTACGACAGTGCAAATACGACAAAAAGCGAGTGATTCTCAAAGATCTCAAGCACAATGATGGTAATTTC ACTGAAAAACAGAAGATAGAATTGAACAAGTTGCTTCAGATTGACTATTACAACCTGACCAAGTTCTACGGCACAGTGAAACTTGATACCATGATCTTCGGGGTGATAGAATACTGTGAGAGAGGATCCCTCCGGGAAGTTTTAAATGACACAATTTCCTACCCTGATGGCACATTCATGGATTGGGAGTTTAAGATCTCTGTCTTGTATGACATTGCTAAGGGAATGTCATATCTGCACTCCAGTAAGA CAGAAGTCCATGGTCGTCTGAAATCTACCAACTGCGTAGTGGACAGTAGAATGGTGGTGAAGATCACTGATTTTGGCTGCAATTCCATTTTACCTCCAAAAAAGGACCTGTGGACAGCTCCAGAGCACCTCCGCCAAGCCAACATCTCTCAGAAAGGGAGATGTGTACAGCTATGGGATCATCGCACAGGAGATCATTCTGCGGAAAGAAACCTTCTACACTTTGAGCTGTCGGGACCGGAATGAGAAGAT TTTCAGAGTGGAAAATTCCAATGGAATGAAACCCTTCCGCCCAGATTTATTCTTGGAAACAGCAGAGGAAAAAGAGCTAGAAGTGTACCTACTTGTAAAAAACTGTTGGGAGGAAGATCCAGAAAAGAGACCAGATTTCAAAAAAATTGAGACTACACTTGCCAAGATTTTGGACTTTTTCATGACCAAAAAAATGAAAGCTATATGGATACCTTGATCCGACGTCTACAGCTATATTCTCGAAACCTGGAACATCT GGTAGAGGAAAGGACACAGCTGTACAAGGCAGAGAGGGACAGGGCTGACAGACTTAACTTTATGTTGCTTCCAAGGCTAGTGGTAAAGTCTCTGAAGGAGAAAGGCTTTGTGGAGCCGGAACTATATGAGGAAGTTACAATCTACTTCAGTGACATTGTAGGTTTCACTACTATCTGCAAATACAGCACCCCCATGGAAGTGGTGGACATGCTTAATGACATCTATAAGAGTTTTGACCACATTGTTGATCAT CATGATGTCTACAAGGTGGAAACCATCGGTGATGCGTACATGGTGGCTAGTGGTTTGCCTAAGAGAAATGGCAATCGGCATGCAATAGACATTGCCAAGATGGCCTTGGAAATCCTCAGCTTCATGGGGACCTTTGAGCTGGAGCATCTTCCTGGCCTCCCAATATGGATTCGCATTGGAGTTCACTCTGGTCCCTGTGCTGCTGGAGTTGTGGGAATCAAGATGCCTCGTTATTGTCTATTTGGAG ATACGGTCAACACAGCCTCTAGGATGGAATCCACTGGCCTCCCTTTGAGAATTCACGTGAGTGGCTCCACCATAGCCATCCTGAAGAGAACTGAGTGCCAGTTCCTTTATGAAGTGAGAGGAGAAACATACTTAAAGGGAAGAGGAAATGAGACTACCTACTGGCTGACTGGGATGAAGGACCAGAAATTCAACCTGCCAACCCCTCCTACTGTGGAGAATCAACAGCGTTTGCAAGCAGAATTTTCAGACATGA TTGCCAACTCTTTACAGAAAAGACAGGCAGCAGGGATAAGAAGCCAAAAACCCAGACGGGTAGCCAGCTATAAAAAAGGCACTCTGGAATACTTGCAGCTGAATACCACAGACAAGGAGAGCACCTATTTTACGCGTACGCGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATTACAAGGATGACGACGATAAG 226 Full size GUCY2C cynomolgus macaque SQVSQNCHNGSYEISVLMMDNSAFAEPLENVEDAVNEGLEIVRGRLQNAGLNVTVNASFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAKRMGCVLMGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLTYFLVNFWKTNDLPFKTYSWSTSYVYKNGTESEDCFWYLNALEASVSYFSHELSFKLVLRQDK EFQDILMDHNRKSNVIVMCGDPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTQSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKTFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNQTNPVDMSPTFTWKNSKLPNDITDRGPQILMIAVFT LTGAVVLLLLVALLMLRKYKKDYELRQKKWSHIPPENIFPLETNETNHVSLKIDDDKRRDTIQRLRQCKYDKKRVILKDLKHNDGNFTEKQKIELNKLLQIDYYNLTKFYGTVKLDTMIFGVIEYCERGSLREVLNDTISYPDGTFMDWEFKISVLYDIAKGMSYLHSSKTEVHGRLKSTNCVVDSRMV VKITDFGCNSILPPKKDLWTAPEHLRQANVSQKGDVYSYGIIAQEIILRKETFYTSSCRDRNEKIFRVENSNGMKPFRPDLFLETAEEKELEVYLLVKSCWEEDPEKRPDFKKIETTLAKIFGLFHDQKNESYMDTLIRRLQLYSRNLEHLVEERTQLYKAERDRADRLNFMLLPRLVVKSLKEKGFVEPELYEEVTIYF SDIVGFTTICKYSTPMEVVDMLNDIYKSFDHIVDHHDVYKVETIGDAYMVASGLPKRNGNRHAIDIAKMALEILSFMGTFELEHLPGLPIWIRIGVHSGPCAAGVVGIKMPRYCLFGDTVNTASRMESTGLPLRIHVSGSTIAILKRTECQFLYEVRGETYLKGRGNETTYWLTGMKDQKFNLPTPPTVENQQRLQAEFSDMIANSLQ KRQAAGIRSQKPRRVASYKKGTLEYLQLNTTDKESTYFTRTRPLEQKLISEEDLAANDILDYKDDDDK 227 Full size GUCY2C cynomolgus macaque tcacaggtgagtcagaactgccacaatggcagctatgaaatcagcgtcctgatgatggacaactcagcctttgcagagcccctggaaaacgtggaagatgcggtgaatgaggggctggaaatagtgagaggacgtctgcaaaacgctggcctaaatgtgactgtgaatgcttctttcatgtattcggatggtctgattcat aactccggcgactgccggagcagcacctgtgaaggccttgacctactcaggaaaatttcaaatgcaaaacggatgggctgtgtcctcatggggccctcatgtacatactccaccttccagatgtaccttgacacagaattgagctaccccatgatctcagctggaagttttggattgtcatgtgactataaagaaaccttaaccaggctgatgtct ccagctagaaagttgacatacttcttggttaacttttggaaaaccaatgatctacccttcaaaacttattcctggagcacttcgtatgtttacaagaatggtacggagtccgaggactgtttctggtaccttaacgctctggaggccagtgtttcctatttctcccacgaactcagttttaagttggtgttaagacaagataaggagtt tcaggatatcttaatggaccacaacaggaaaagcaatgtgattgttatgtgtggtgatccaGagttcctctacaagttgaagggtgaccgagcagtggctgaagacattgtcattattctagtggatcttttcaatgaccagtactttgaggacaatgtcacagcccctgactatatgaaaaatgtccttgttctgacgcag tctcctggcaattccctcctaaatagctctttctccaggaatctatccccaacaaaacgagactttgctcttgcctatttgaatggaatcctgctctttggacatatgctaaagacatttcttgaaaatggagaaaatattaccacccccaaatttgctcatgcttttcaggaatctcacttttgaagggtatgacggtccagtgaccttggatg actggggggatgtggacagtaccatggtgcttctgtatacgtctgtggacaccaagaaatacaaggttcttttgacctatgatacccacgtaaatcagaccaaccctgtggatatgagccccacattcacttggaagaactctaaacttcctaatgatattacagaccggggccctcagatcctgatgattgcagtcttcaccctcaccggagctgtggt gctgctcctgcttgtcgctctcctgatgctcagaaaatataaaaaagattatgaacttcgtcagaaaaaatggtcccacattcctcctgaaaatatctttcctctggagaccaatgagaccaatcacgttagcctgaagatcgatgatgacaaaagacgagatacaatccagagactacgacagtgcaaatacgacaaaaagcgag tgattctcaaagatctcaagcacaatgatggtaatttcactgaaaaacagaagatagaattgaacaagttgcttcagattgactattacaacctgaccaagttctatggcaccgtgaaacttgataccatgatcttcggggtgatagaatactgtgagagaggatccctccgggaagttttaaatgacacaatttcctaccctgatggcacattcatggattgggagt ttaagatctctgtcctgtatgacattgctaagggaatgtcatatctgcactccagtaagacagaagtccatggtcgtctgaaatctaccaactgcgtagtggacagtagaatggtggtgaagatcactgattttggctgcaattccatttacctccaaaaaaagacctgtggacagctccagagcacctccgccaagccaacgtctctcagaaagg agatgtgtacagctacgggatcatcgcacaggagatcatcctgcggaaagaaaccttctacacttcgagctgtcgagaccggaacgagaagattttcagagtggaaaattccaatggaatgaaacccttccgtccagatttattcttggaaacggcagaggaaaaagagctagaagtgtacctacttgtaaaaagctgttgggaagaagat ccagaaaagagaccagatttcaaaaaaattgagactacacttgccaagatatttggactttttcatgaccaaaaaaatgaaagctatatggataccttgatccgacgtctacagctatattctcgaaacctggaacatctggtagaggaaaggacacagctatacaaggcagagagggacagggctgacagacttaactttatgttgcttccaaggctagtgg taaagtctctgaaggagaaaggctttgtagagccggaactatatgaggaagttacaatctacttcagtgacattgtaggtttcactactatctgcaaatacagcacccccatggaagtggtggacatgcttaatgacatctataagagttttgaccacattgttgatcatcatgatgtctacaaggtggaaaccattggtgatgcctacatggtggctag tggtttgcctaagagaaatggcaatcggcatgcaatagacattgccaagatggccttggaaatcctcagcttcatggggacctttgagctggagcatcttcctggcctcccaatatggattcgcattggcgttcactctggtccctgcgctgctggagttgtgggaatcaagatgcctcgttattgtctatttggagatacagtcaacacag cctctaggatggaatccactggcctccctttgaggattcatgtgagtggctccaccatagccattctgaagagaactgagtgccagttcctgtatgaagtgagaggagaaacgtacttaaagggaagaggaaatgagactacctactggctgaccgggatgaaggaccagaaattcaacctgccaacccctcctactgtggagaatcaacagcgtttgcaagcagaattt tcagacatgattgccaactctttacagaaaagacaggcggcagggataagaagccaaaaacccagacgagtagccagctataaaaaaggcactctggaatacttgcaactgaataccacggacaaggagagcacctattttACGCGTACTCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATTACAAGGATGACGACGATAAG 228 Full size GUCY2C mouse VFWASQVRQNCRNGSYEISVLMMDNSAYKEPMQNLREAVEEGLDIVRKRLREADLNVTVNATFIYSDGLIHKSGDCRSSTCEGLDLLREITRDHKMGCALMGPSCTYSTFQMYLDTELNYPMISAGSYGLSCDYKETLTRILPPARKLMYFLVDFWKVNNASFKPFSWNSSYVYKNGSEPEDCFWYLNALEAGVSYFSEVLNFK DVLRRSEQFQEILTGHNRKSNVIVMCGTPESFYDVKDLQVAEDTVVILVDLFSNHYFEENTTAPEYMDNVLVLTLPSEQSTSNTSVAERFSSGRSDFSLAYLEGTLLFGHMLQTFLENGENVTGPKFARAFRNLTFQGFAGPVTLDDSGDIDNIMSLLYVSLDTRKYKVLMKYDTHKNKTIPVAENPNFIWKNHKLPNDVPGLGPQILMIAV FTLTGILVVLLLIALLVLRKYRRDHALRQKKWSHIPSENIFPLETNETNHISLKIDDDRRRDTIQRVRQCKYDKKKVILKDLKHSDGNFSEKQKIDLNKLLQSDYYNLTKFYGTVKLDTRIFGVVEYCERGSLREVLNDTISYPDGTFMDWEFKISVLNDIAKGMSYLHSSKIEVHGRLKSTNCVVDSRMVVKIT DFGCNSILPPKKDLWTAPEHLRQATISQKGDVYSFAIIAQEIILRKETFYTLSCRDHNEKIFRVENSYGKPFRPDLFLETADEKELEVYLLVKSCWEEDPEKRPDFKKIESTLAKIFGLFHDQKNESYMDTLIRRLQLYSRNLEHLVEERTQLYKAERDRADHLNFMLLPRLVVKSLKEKGIVEPELYEEVTIYFSDIVGFTT ICKYSTPMEVVDMLNDIYKSFDQIVDHHDVYKVETIGDAYVVASGLPMRNGNRHAVDISKMALDILSFIGTFELEHLPGLPVWIRIGVHSGPCAAGVVGIKMPRYCLFGDTVNTASRMESTGLPLRIHMSSSTITILKRTDCQFLYEVRGETYLKGRGTETTYWLTGMKDQEYNLPSPPTVENQQRLQTEFSDMIVSALQKRQASG KKSRRPTRVASYKKGFLEYMQLNNSDHDSTYFTRTRPLEQKLISEEDLAANDILDYKDDDDK 229 Full size GUCY2C mouse GTGTTCTGGGCCTCTCAGGTGAGGCAGAACTGCCGCAATGGCAGCTACGAGATCAGCGTCCTGATGATGGACAACTCAGCCTACAAAGAACCTATGCAAAACCTGAGGGAGGCTGTGGAGGAAGGACTGGACATAGTGCGAAAGCGCCTGCGTGAAGCCGACCTAAATGTGACTGTGAACGCGACTTTCATCTACTCCGACGGTCTGATTCATAAGTCAGGTGACTGCCGGAGCAGCACCTGTGAAGGCCT TGACCTACTCAGGGAGATTACAAGAGATCATAAGATGGGCTGCGCCCTCATGGGGCCCTCGTGCACGTATTCCACCTTCCAGATGTACCTCGACACAGAGTTGAACTATCCCATGATTTCCGCTGGAAGTTATGGATTGTCCTGTGACTATAAGGAAACCCTAACCAGGATCCTGCCTCCAGCCAGGAAGCTGATGTACTTCTTGGTCGATTTCTGGAAAGTCAACAATGCATCTTTCAAACCCTTTTCCTG GAACTCTTCGTATGTTTACAAGAATGGATCGGAACCTGAAGATTGTTTCTGGTACCTCAATGCTCTGGAGGCTGGGGTGTCCTATTTTTCTGAGGTGCTCAACTTCAAGGATGTACTGAGACGCAGCGAACAGTTCCAGGAAATCTTAACAGGCCATAACAGAAAGAGCAATGTGATTGTTATGTGTGGCACGCCAGAAAGCTTCTATGATGTGAAAGGTGACCTCCAAGTGGCTGAAGATACTGTT GTCATCCTGGTAGATCTGTTCAGTAACCATTACTTTGAGGAGAACACCACAGCTCCTGAGTATATGGACAATGTCCTCGTCCTGACGCTGCCGTCTGAACAGTCCACCTCAAACACCTCTGTCGCCGAGAGGTTTTCATCGGGGAGAAGTGACTTTTCTCTCGCTTACTTGGAGGGAACCTTGCTATTTGGACACATGCTGCAGACGTTTCTTGAAAATGGAGAAAATGTCACGGGTCCCAAGTTTGCT CGTGCATTCAGGAATCTCACTTTTCAAGGCTTTGCAGGACCTGTGACTCTGGATGACAGTGGGGACATTGACAACATTATGTCCCTTCTGTATGTGTCTCTGGATACCAGGAAATACAAGGTTCTTATGAAGTATGACACCCACAAAAACAAAACTATTCCGGTGGCTGAGAACCCCAACTTCATCTGGAAGAACCACAAGCTCCCCCAATGACGTTCCTGGGCTGGGCCCTCAAATCCTGATGATTGCCG TCTTCACGCTCACGGGGATCCTGGTAGTTCTGCTGCTGATTGCCCTCCTCGTGCTGAGAAAATACAGAAGAGATCATGCACTTCGACAGAAGAAATGGTCCCACATTCCTTCTGAAAACATCTTTCCTCTGGAGACCAACGAGACCAACCACATCAGCCTGAAGATTGACGATGACAGGAGACGAGACACAATCCAGAGAGTGCGACAGTGCAAATACGACAAGAAGAAAGTGATTCTGAAAGACCTCAAGCAC AGCGACGGGAACTTCAGTGAGAAGCAGAAGATAGACCTGAACAAGTTGCTGCAGTCTGACTACTACAACCTGACTAAGTTCTACGGCACCGTGAAGCTGGACACCAGGATCTTTGGGGTGGTTGAGTACTGCGAGAGGGGATCCCTCCGGGAAGTGTTAAACGACACAATTTCCTACCCTGACGGCACGTTCATGGATTGGGAGTTTAAGATCTCTGTCTTAAATGACATCGCTAAGGGGATGTCCTAC CTGCACTCCAGTAAGATTGAAGTCCACGGGCGTCTCAAATCCACCAACTGCGTGGTGGACAGCCGCATGGTGGTGAAGATCACCGACTTTGGGTGCAATTCCATCCTGCCTCCAAAAAAAGACCTGTGGACGGCCCCGGAGCACCTGCGCCAGGCCACCATCTCTCAGAAAGGAGACGTGTACAGCTTCGCCATCATTGCCCAGGAGATCATCCTCCGTAAGGAGACTTTTTACACGGCTGAGCTGTCGAT CACAATGAGAAGATTTTCAGAGTGGAAAATTCATACGGGAAACCTTTCCGCCCAGACCTCTTCCTGGAGACTGCAGATGAGAAGGAGCTGGAGGTCTATCTACTTGTCAAAAGCTGTTGGGAGGAGGATCCAGAAAAGAGGCCAGATTTCAAGAAAATCGAGAGCACACTGGCCAAGATATTTGGCCTTTTCCATGACCAGAAAAACGAGTCTTACATGGACACCTTGATCCGACGTCTCCAGCTGTACTCTCGAAA CCTGGAACATCTGGTGGAGGAAAGGACTCAGCTGTACAAGGCGGAGAGGGACAGGGCTGACCACCTTAACTTCATGCTCCTCCCACGGCTGGTGGTAAAGTCACTGAAGGAGAAAGGCATCGTGGAGCCAGAGCTGTACGAAGAAGTCACAATCTACTTCAGTGACATTGTGGGCTTCACCACCATCTGCAAGTATAGCACGCCCATGGAGGTGGTGGACATGCTCAACGACATCTACAAGAGCTTTGA CCAGATTGTGGACCACCATGACGTCTACAAGGTAGAAACCATCGGTGACGCCTACGTGGTGGCCAGCGGTCTGCCTATGAGAAACGGCAACCGACACGCGGTAGACATTTCCAAGATGGCCTTGGACATCCTCAGCTTCATAGGGACCTTTGAGTTGGAGCATCTCCCTGGCCTCCCCGTGTGGATCCGCATTGGAGTTCATTCTGGGCCCTGCGCTGCTGGTGTTGTGGGGATCAAGATGCCTCGCTATT GCCTGTTTGGAGACACTGTCAACACTGCCTCCAGGATGGAATCCACCGGCCTCCCCTTGAGGATTCACATGAGCAGCTCCACCATAACCATCCTGAAGAGAACGGATTGCCAGTTCCTGTATGAAGTGAGGGGAGAAACCTACTTAAAGGGAAGAGGGACAGACCACATACTGGCTGACTGGGATGAAGGACCAAGAATACAACCTGCCATCCCCACCGACAGTGGAGAACCAACAGCGTCTGCAGACT GAGTTCTCAGACATGATCGTTAGCGCCTTACAGAAAAGACAGGCCTCGGGCAAGAAGAGCCGGAGGCCCACTCGGGTGGCCAGCTACAAGAAAGGCTTTCTGGAATACATGCAGCTGAACAATTCAGACCACGATAGCACCTATTTTACGCGTACGCGGCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATTACAAGGATGACGACGATAAG 230 Human ECD GUCY2C-muIgG2a SQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQD KEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQGGGGSENLY FQGGGGSGGGGSEPRGPTIKPCPPCKCPAPNLEGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKAFACAVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSD GSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGGGPPDYKDDDDK 231 Human ECD GUCY2C-muIgG2a ATGGAGACAGACACACTGCTCCTGTGGGTGCTGCTTCTGTGGGTGCCAGGTTCCACTGGAtcacaggtgagtcagaactgccacaatggcagctatgaaatcagcgtcctgatgatgggcaactcagcctttgcagagcccctgaaaaacttggaagatgcggtgaatgaggggctggaaatagtgagaggacgtctgcaaaatgctggccta aatgtgactgtgaacgctactttcatgtattcggatggtctgattcataactcaggcgactgccggagtagcacctgtgaaggcctcgacctactcaggaaaatttcaaatgcacaacggatgggctgtgtcctcatagggccctcatgtacatactccaccttccagatgtaccttgacacagaattgagctaccccatgatctcagctggaagttt tggattgtcatgtgactataaagaaaccttaaccaggctgatgtctccagctagaaagttgatgtacttcttggttaacttttggaaaaccaacgatctgcccttcaaaacttattcctggagcacttcgtatgtttacaagaatggtacagaaactgaggactgtttctggtaccttaatgctctggaggctagcgtttcctatttct cccacgaactcggctttaaggtggtgttaagacaagataaggagtttcaggatatcttaatggaccacaacaggaaaagcaatgtgattattatgtgtggtggtccagagttcctctacaagctgaagggtgaccgagcagtggctgaagacattgtcattattctagtggatcttttcaatgaccagtactttgaggacaatgtcacagcccc tgactatatgaaaaatgtccttgttctgacgctgtctcctgggaattcccttctaaatagctcttttctccaggaatctatcaccaacaaaacgagactttgctcttgcctatttgaatggaatcctgctctttggacatatgctgaagatatttcttgaaaatggagaaaatattaccacccccaaatttgctcatgctttcaggaatct cacttttgaagggtatgacggtccagtgaccttggatgactggggggatgttgacagtaccatggtgcttctgtatacctctgtggacaccaagaaatacaaggttcttttgacctatgatacccacgtaaataagacctatcctgtggatatgagccccacattcacttggaagaactctaaacttcctaatgattacaggccggggccctca gggaggcggaggttccGAGAATTTATACTTCCAGGGCGGAGGCGGTTCCGGCGGCGGAGGAAGCgagccccgcggaccgacaatcaagccctgtcctccatgcaaatgcccagcacctaacctcgaggtggaccatccgtcttcatcttccctccaaagatcaaggatgtactcatgatctccctgagccccatagtcacatgtgtggtggtggatgtgag cgaggatgacccagatgtccagatcagctggtttgtgaacaacgtggaagtacacacagctcagacacaaacccatagagaggattacaacagtactctccgggtggtcagtgccctccccatccagcaccaggactggatgagtggcaaggctttcgcatgcgccgtcaacaacaaagacctcccagcgcccatcgagagaaccatctcaaaacc caaagggtcagtaagagctccacaggtatatgtcttgcctccaccagaagaagagatgactaagaaacaggtcactctgacctgcatggtcacagacttcatgcctgaagacatttacgtggagtggaccaacaacgggaaaacagagctaaactacaagaacactgaaccagtcctggactctgatggttcttacttcatgtacagcaagctgagagtggaa aagaagaactgggtggaaagaaatagctactcctgttcagtggtccacgagggtctgcacaatcaccacacgactaagagcttctcccggactccgggtGGCGGGCCACCCGACTACAAGGACGACGATGACAAA 232 Human cleaved GUCY2C ECD SQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQD KEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQGGGGSENLY FQ 233 Human cleaved GUCY2C ECD tcacaggtgagtcagaactgccacaatggcagctatgaaatcagcgtcctgatgatgggcaactcagcctttgcagagcccctgaaaaacttggaagatgcggtgaatgaggggctggaaatagtgagaggacgtctgcaaaatgctggcctaaatgtgactgtgaacgctactttcatgtattcggatggtctgattcataact caggcgactgccggagtagcacctgtgaaggcctcgacctactcaggaaaatttcaaatgcacaacggatgggctgtgtcctcatagggccctcatgtacatactccaccttccagatgtaccttgacacagaattgagctaccccatgatctcagctggaagttttggattgtcatgtgactataaagaaaccttaaccaggctgatgtctccagctaga aagttgatgtacttcttggttaacttttggaaaaccaacgatctgcccttcaaaacttattcctggagcacttcgtatgtttacaagaatggtacagaaactgaggactgtttctggtaccttaatgctctggaggctagcgtttcctatttctcccacgaactcggctttaaggtggtgttaagacaagataaggagtttcagg atatcttaatggaccacaacaggaaaagcaatgtgattattatgtgtggtggtccagagttcctctacaagctgaagggtgaccgagcagtggctgaagacattgtcattattctagtggatcttttcaatgaccagtactttgaggacaatgtcacagcccctgactatatgaaaaatgtccttgttctgacgcttctcctggga attcccttctaaatagctctttctccaggaatctatcaccaacaaaacgagactttgctcttgcctatttgaatggaatcctgctctttggacatatgctgaagatatttcttgaaaatggagaaaatattaccacccccaaatttgctcatgctttcaggaatctcacttttgaagggtatgacggtccagtgaccttggatgactggggggatg ttgacagtaccatggtgcttctgtatacctctgtggacaccaagaaatacaaggttctttgacctatgatacccacgtaaataagacctatcctgtggatatgagccccacattcacttggaagaactctaaacttcctaatgattacaggccggggccctcagggaggcggaggttccGAGAATTTATACTTCCAG 234 ECD GUCY2C cynomolgus macaque-muIG2a SQVSQNCHNGSYEISVLMMDNSAFAEPLENVEDAVNEGLEIVRGRLQNAGLNVTVNASFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAKRMGCVLMGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLTYFLVNFWKTNDLPFKTYSWSTSYVYKNGTESEDCFWYLNALEASVSYFSHELSFKLVLRQDK EFQDILMDHNRKSNVIVMCGDPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTQSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKTFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNQTNPVDMSPTFTWKNSKLPNDITDRGPQGGGGSEN LYFQGGGGSGGGGSEPRGPTIKPCPPCKCPAPNLEGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKAFACAVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLD SDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGGGPPDYKDDDDK 235 ECD GUCY2C cynomolgus macaque-muIG2a tcacaggtgagtcagaactgccacaatggcagctatgaaatcagcgtcctgatgatggacaactcagcctttgcagagcccctggaaaacgtggaagatgcggtgaatgaggggctggaaatagtgagaggacgtctgcaaaacgctggcctaaatgtgactgtgaatgcttctttcatgtattcggatggtctgattcat aactccggcgactgccggagcagcacctgtgaaggccttgacctactcaggaaaatttcaaatgcaaaacggatgggctgtgtcctcatggggccctcatgtacatactccaccttccagatgtaccttgacacagaattgagctaccccatgatctcagctggaagttttggattgtcatgtgactataaagaaaccttaaccaggctgatgtct ccagctagaaagttgacatacttcttggttaacttttggaaaaccaatgatctacccttcaaaacttattcctggagcacttcgtatgtttacaagaatggtacggagtccgaggactgtttctggtaccttaacgctctggaggccagtgtttcctatttctcccacgaactcagttttaagttggtgttaagacaagataaggagtt tcaggatatcttaatggaccacaacaggaaaagcaatgtgattgttatgtgtggtgatccaGagttcctctacaagttgaagggtgaccgagcagtggctgaagacattgtcattattctagtggatcttttcaatgaccagtactttgaggacaatgtcacagcccctgactatatgaaaaatgtccttgttctgacgcag tctcctgggaattccctcctaaatagctctttctccaggaatctatccccaacaaaacgagactttgctcttgcctatttgaatggaatcctgctctttggacatatgctaaagacatttcttgaaaatggagaaaatattaccacccccaaatttgctcatgcttttcaggaatctcacttttgaagggtatgacggtccagtgaccttggatg actggggggatgtggacagtaccatggtgcttctgtatacgtctgtggacaccaagaaatacaaggttcttttgacctatgatacccacgtaaatcagaccaaccctgtggatatgagccccacattcacttggaagaactctaaacttcctaatgatattacagaccggggccctcagggaggcggaggttccGAGAATTTATACTTCCAGGGCGGAGGCGGTTCC GGCGGCGGAGGAAGCgagccccgcggaccgacaatcaagccctgtcctccatgcaaatgcccagcacctaacctcgagggtggaccatccgtcttcatcttccctccaaagatcaaggatgtactcatgatctccctgagccccatagtcacatgtgtggtggtggatgtgagcgaggatgacccagatgtccagatcagctggtttgtgaaca acgtggaagtacacacagctcagacacaaacccatagagaggattacaacagtactctccgggtggtcagtgccctccccatccagcaccaggactggatgagtggcaaggctttcgcatgcgccgtcaacaacaaagacctcccagcgcccatcgagaaccatctcaaaacccaaagggtcagtaagagctccacaggtatatgtcttgcctccacc agaagaagagatgactaagaaacaggtcactctgacctgcatggtcacagacttcatgcctgaagacatttacgtggagtggaccaacaacgggaaaacagagctaaactacaagaacactgaaccagtcctggactctgatggttcttacttcatgtacagcaagctgagagtggaaaagaagaactgggtggaaagaaatagctactcctgttcagtggt ccacgaggtctgcacaatcaccacacgactaagagcttctcccggactccgggtGGCGGGCCACCCGACTACAAGGACGACGATGACAAA 236 Cynomolgus macaque split ECD GUCY2C SQVSQNCHNGSYEISVLMMDNSAFAEPLENVEDAVNEGLEIVRGRLQNAGLNVTVNASFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAKRMGCVLMGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLTYFLVNFWKTNDLPFKTYSWSTSYVYKNGTESEDCFWYLNALEASVSYFSHELSFKLVLRQDK EFQDILMDHNRKSNVIVMCGDPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTQSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKTFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNQTNPVDMSPTFTWKNSKLPNDITDRGPQGGGGSEN LYFQ 237 Cynomolgus macaque split ECD GUCY2C tcacaggtgagtcagaactgccacaatggcagctatgaaatcagcgtcctgatgatggacaactcagcctttgcagagcccctggaaaacgtggaagatgcggtgaatgaggggctggaaatagtgagaggacgtctgcaaaacgctggcctaaatgtgactgtgaatgcttctttcatgtattcggatggtctgattcat aactccggcgactgccggagcagcacctgtgaaggccttgacctactcaggaaaatttcaaatgcaaaacggatgggctgtgtcctcatggggccctcatgtacatactccaccttccagatgtaccttgacacagaattgagctaccccatgatctcagctggaagttttggattgtcatgtgactataaagaaaccttaaccaggctgatgtct ccagctagaaagttgacatacttcttggttaacttttggaaaaccaatgatctacccttcaaaacttattcctggagcacttcgtatgtttacaagaatggtacggagtccgaggactgtttctggtaccttaacgctctggaggccagtgtttcctatttctcccacgaactcagttttaagttggtgttaagacaagataaggagtt tcaggatatcttaatggaccacaacaggaaaagcaatgtgattgttatgtgtggtgatccaGagttcctctacaagttgaagggtgaccgagcagtggctgaagacattgtcattattctagtggatcttttcaatgaccagtactttgaggacaatgtcacagcccctgactatatgaaaaatgtccttgttctgacgcag tctcctgggaattccctcctaaatagctctttctccaggaatctatccccaacaaaacgagactttgctcttgcctatttgaatggaatcctgctctttggacatatgctaaagacatttcttgaaaatggagaaaatattaccacccccaaatttgctcatgcttttcaggaatctcacttttgaagggtatgacggtccagtgaccttggatg actggggggatgtggacagtaccatggtgcttctgtatacgtctgtggacaccaagaaatacaaggttcttttgacctatgatacccacgtaaatcagaccaaccctgtggatatgagccccacattcacttggaagaactctaaacttcctaatgatattacagaccggggccctcagggaggcggaggttccGAGAATTTATACTTCCAG 238 Mouse ECD GUCY2C-muIgG2a VFWASQVRQNCRNGSYEISVLMMDNSAYKEPMQNLREAVEEGLDIVRKRLREADLNVTVNATFIYSDGLIHKSGDCRSSTCEGLDLLREITRDHKMGCALMGPSCTYSTFQMYLDTELNYPMISAGSYGLSCDYKETLTRILPPARKLMYFLVDFWKVNNASFKPFSWNSSYVYKNGSEPEDCFWYLNALEAGVSYFSEVLNFK DVLRRSEQFQEILTGHNRKSNVIVMCGTPESFYDVKDLQVAEDTVVILVDLFSNHYFEENTTAPEYMDNVLVLTLPSEQSTSNTSVAERFSSGRSDFSLAYLEGTLLFGHMLQTFLENGENVTGPKFARAFRNLTFQGFAGPVTLDDSGDIDNIMSLLYVSLDTRKYKVLMKYDTHKNKTIPVAENPNFIWKNHKLPNDVPGLGPQILMG GSENLYFQGGGGSGGGGSEPRGPTIKPCPPCKCPAPNLEGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKAFACAVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEP VLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGHHHHHH 239 Mouse ECD GUCY2C-muIgG2a gtgttctgggcctctcaggtgaggcagaactgccgcaatggcagctacgagatcagcgtcctgatgatggacaactcagcctacaaagaacctatgcaaaacctgagggaggctgtggaggaaggactggacatagtgcgaaagcgcctgcgtgaagccgacctaaatgtgactgtgaacgcgactttcatctactccgacggt ctgattcataagtcaggtgactgccggagcagcacctgtgaaggccttgacctactcagggattacaagagatcataagatgggctgcgccctcatggggccctcgtgcacgtattccaccttccagatgtacctcgacacagagttgaactatcccatgatttccgctggaagttatggattgtcctgtgactataaggaaaccctaaccagg attctgcctccagccaggaagctgatgtacttcttggtcgatttctggaaagtcaacaatgcatctttcaaacccttttcctggaactcttcgtatgtttacaagaatggatcggaacctgaagattgtttctggtacctcaatgctctggaggctggggtgtcctatttttctgaggtgctcaacttcaaggatgtactgagacg cagcgaacagttccaggaaatcttaacaggccataacagaaagagcaatgtgattgttatgtgtggcacgccagaaagcttctatgatgtgaaaggtgacctccaagtggctgaagatactgttgtcatcctggtagatctgttcagtaaccattactttgaggagaacaccacagctcctgagtatatggacaatgtcctcgtcctgac gctgccgtctgaacagtccacctcaaacacctctgtcgccgagaggttttcatcggggagaagtgacttttctctcgcttacttggagggaaccttgctatttggacacatgctgcagacgtttcttgaaaatggagaaaatgtcacgggtcccaagtttgctcgtgcattcaggaatctcacttttcaaggcttt gcaggacctgtgactctggatgacagtggggacattgacaacattatgtcccttctgtatgtgtctctggataccaggaaatacaaggttcttatgaagtatgacacccacaaaaacaaaactattccggtggctgagaaccccaacttcatctggaagaaccacaagctccccaatgacgttcctgggctgggccctcaaatcctgatgggaggcggagg ttccGAGAATTTATACTTCCAGGGCGGAGGCGGTTCCGGCGGCGGAGGAAGCgagccccgcggaccgacaatcaagccctgtcctccatgcaaatgcccagcacctaacctcgaggtggaccatccgtcttcatcttccctccaaagatcaaggatgtactcatgatctccctgagccccatagtcacatgtgtggtggtggatgtgagcgaggatgaccc agatgtccagatcagctggtttgtgaacaacgtggaagtacacacagctcagacacaaacccatagagaggattacaacagtactctccgggtggtcagtgccctccccatccagcaccaggactggatgagtggcaaggctttcgcatgcgccgtcaacaacaaagacctcccagcgcccatcgagaaccatctcaaaacccaaagggtcag taagagctccacaggtatatgtcttgcctccaccagaagaagagatgactaagaaacaggtcactctgacctgcatggtcacagacttcatgcctgaagacatttacgtggagtggaccaacaacgggaaaacagagctaaactacaagaacactgaaccagtcctggactctgatggttcttacttcatgtacagcaagctgagagtggaaaagaagaactgggt ggaaagaaatagctactcctgttcagtggtccacgaggtctgcacaatcaccacacgactaagagcttctcccggactccgggtCACCATCACCATCACCAT 240 Mouse GUCY2C split ECD VFWASQVRQNCRNGSYEISVLMMDNSAYKEPMQNLREAVEEGLDIVRKRLREADLNVTVNATFIYSDGLIHKSGDCRSSTCEGLDLLREITRDHKMGCALMGPSCTYSTFQMYLDTELNYPMISAGSYGLSCDYKETLTRILPPARKLMYFLVDFWKVNNASFKPFSWNSSYVYKNGSEPEDCFWYLNALEAGVSYFSEVLNFK DVLRRSEQFQEILTGHNRKSNVIVMCGTPESFYDVKDLQVAEDTVVILVDLFSNHYFEENTTAPEYMDNVLVLTLPSEQSTSNTSVAERFSSGRSDFSLAYLEGTLLFGHMLQTFLENGENVTGPKFARAFRNLTFQGFAGPVTLDDSGDIDNIMSLLYVSLDTRKYKVLMKYDTHKNKTIPVAENPNFIWKNHKLPNDVPGLGPQILMG GSENLYFQ 241 Mouse GUCY2C split ECD gtgttctgggcctctcaggtgaggcagaactgccgcaatggcagctacgagatcagcgtcctgatgatggacaactcagcctacaaagaacctatgcaaaacctgagggaggctgtggaggaaggactggacatagtgcgaaagcgcctgcgtgaagccgacctaaatgtgactgtgaacgcgactttcatctactccgacggt ctgattcataagtcaggtgactgccggagcagcacctgtgaaggccttgacctactcagggattacaagagatcataagatgggctgcgccctcatggggccctcgtgcacgtattccaccttccagatgtacctcgacacagagttgaactatcccatgatttccgctggaagttatggattgtcctgtgactataaggaaaccctaaccagg attctgcctccagccaggaagctgatgtacttcttggtcgatttctggaaagtcaacaatgcatctttcaaacccttttcctggaactcttcgtatgtttacaagaatggatcggaacctgaagattgtttctggtacctcaatgctctggaggctggggtgtcctatttttctgaggtgctcaacttcaaggatgtactgagacg cagcgaacagttccaggaaatcttaacaggccataacagaaagagcaatgtgattgttatgtgtggcacgccagaaagcttctatgatgtgaaaggtgacctccaagtggctgaagatactgttgtcatcctggtagatctgttcagtaaccattactttgaggagaacaccacagctcctgagtatatggacaatgtcctcgtcctgac gctgccgtctgaacagtccacctcaaacacctctgtcgccgagaggttttcatcggggagaagtgacttttctctcgcttacttggagggaaccttgctatttggacacatgctgcagacgtttcttgaaaatggagaaaatgtcacgggtcccaagtttgctcgtgcattcaggaatctcacttttcaaggcttt gcaggacctgtgactctggatgacagtggggacattgacaacattatgtcccttctgtatgtgtctctggataccaggaaatacaaggttcttatgaagtatgacacccacaaaaacaaaactattccggtggctgagaaccccaacttcatctggaagaaccacaagctccccaatgacgttcctgggctgggccctcaaatcctgatgggaggcggagg ttccGAGAATTTATACTTCCAG 242 Human CD3 epsilon delta mqsgthwrvlglcllsvgvwgqdgneemggitqtpykvsisgttviltcpqypgseilwqhndkniggdeddknigsdedhlslkefseleqsgyyvcyprgskpedanfylylrarggggsggggsggggspieeledrvfvncntsitwvegtvgtllsditrldlgkrildprgiyrcngtdiy kdkestvqvhyrmcqscveldhhhhhh 243 Human CD3 epsilon delta atgcaatccggtacgcactggagagtcttgggtctgtgccttttgtctgttggcgtatgggggcaagacgggaacgaagaaatgggaggcattacacaaacaccatacaaggtatcaattagcggcacgacggttatactgacatgtccacaatatccaggcagcgaaattctgtggcagcacaatgacaagaatattgggggatgaagacgacaa aaatatcggtagcgacgaggaccatctgtctctgaaggaattttcagaacttgaacaatctggctattatgtgtgctacccgcgaggcagcaaaccggaagatgcaaacttttacctttatctgagagcaaggggcggcggaggctctgggggcggaggcagcggcggaggaggatcaccaatcgaggaattggaagatagggtattcgtaaattg taacaccagcattacatgggtggaagggaccgttggaactctcctgagcgatatcacacgactggatcttggtaaacgaatcctggacccacgcggaatctatagatgtaacggaactgatatttacaaagacaaagaatctactgtgcaagttcactaccgaatgtgtcaatcatgcgttgaactcgatcaccaccaccatcatcac 244 CD3 epsilon delta cynomolgus monkey mqsgthwrvlglcllsvgvwgqdgneemgsitqtpyqvsisgttviltcsqhlgseaqwqhngknkgdsgdqlflpefsemeqsgyyvcyprgsnpedashhlylkarggggsggggsggggspveeledrvfvkcntsvtwvegtvgtlltnntrldlgkrildprgiyrcngtdiykdkes avqvhyrmcqncveldhhhhhh 245 CD3 epsilon delta cynomolgus monkey atgcaaagcggaactcattggcgcgtcctgggactctgtctgctctccgtgggagtctggggacaagatggaaacgaagagatgggaagcattacacaaacaccataccaagtctccattagcggcactaccgtcattctgacatgttcccaacatctgggcagcgaagctcaatggcagcacaatggaaagaataagggcgatagcggagaccaactgttt ctgccagaatttagcgaaatggagcaatccggctattacgtgtgctacccacgcggcagcaaccctgaagatgctagccatcacctctatctgaaggctcgcggaggcggaggcagcggcggcggaggatccggcggaggcggaagcccagtcgaggaactggaagatcgcgtcttcgtgaagtgtaacaccagcgtcacatgggtgg aaggcaccgtcggaactctctcctgactaacaacacacgcctggatctcggaaaacgcatcctggacccacgcggaatctatagatgtaacggaactgatatttacaaagacaaagaatccgctgtgcaagtccactaccgcatgtgtcaaaactgtgtcgaactggatcatcaccatcaccatcac 246 GUCY2C-1608_protrusion gacatccagctgacccagtctccatcctccctgtctgcatctgtaggagacagagtcaccatcacttgcagagccagtgaaagtgttgattattatggcagtagtttattgcagtggtatcagcagaaaccagggaaagcccctaagctcctgatctatgctgcatccaaactagcttctggggtcccatcaaggttcagtggcagtggatct gggacagatttcactctcaccatcagcagtctgcaacctgaagattttgcaacttactactgtcagcaaactcggaaagcgtatacgtttggccaggggaccaagctggagatcaaaggtggaggtagcgggggcggcggggaagttcaactcgttgagtctggcgggggattggttcaacccggtggaagccttagattgtcatgtgcc gcctccggctttacatttagcgactattacatgacctgggtgagacaagctccaggcaaaggacttgaatgggtggcctttatcagaaatcaggcccgcggctacacaagcgaccataatccctccgtgaaaggaagatttaccatcagccgggacaatgctaaaaattcactttaccttcaaatgaactctcttagagccgaggacaccgccgtatactactg cgcaagagatagaccaagttattacgtcctggattactggggccagggaacaaccgtcaccgtgtcttctggatgcccaccgtgcccagcacctgaagccgctggggcaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtca agttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgaga accacaggtgtgcaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgtggtgcctggtcaaaggcttctatcccagcgacatcgccgtggagtggggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctatagcaagctcaccgtggacaagagcagg tggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtcccccggaaag 247 GUCY2C-1608_cavity gacatccagatgacccagtccccctcttctctgtctgcctctgtgggcgacagagtgaccatcacctgcacaagctcacagtcactgtttaatgtccgcagccagaaaaactatcttgcgtggtatcagcagaagcctggcaaggctcccaagctgctgatctactgggccagtacacgagaatccggcgtgccttccagattct ccggctctggctctggcaccgatttcaccctgaccatctcctccctccagcctgaggatttcgccacctactactgcaaacagtcttacgaccttttcacttttggcggcggaacaaaggtggagatcaagggcggaggtggatctggcggcggaggcgaggtgcagctggtggagtctgggggaggcttggtccagcctggggggtccctga gactctcctgtgcagcctctggcttcaccttcagcagctactggatgcactgggtccgccaggctccagggaaggggctggagtggattggagagattaaacctagcaacgaacttactaacgtccatgaaaagttcaaggaccgattcaccatctccgtggacaaggccaagaactcagcctatctgcaaatgaacagcctgagagccgaggacacggctgt gtattactgtacaagaacgattacgacgacggagggatactggttcttcgatgtctggggccaagggacactggtcaccgtctcttcaggatgtccaccgtgcccagcacctgaagccgctggggcaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgag ccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaag ccaaagggcagccccgagaaccacaggtgtacaccctgcccccatgccgggaggagatgaccaagaaccaggtcagcctgtcctgcgcggtcaaaggcttctatcccagcgacatcgccgtggagtggggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctcgttagcaagct caccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtcccccggaaag 248 GUCY2C-0250_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 249 GUCY2C-0250_cavity DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSRKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 250 GUCY2C-0250_monomer DIQLTQSPSSLSASVGDRVTITCRASESVDYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKVYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNRARGYTSDHNPSVKGRFTISRDNAKNSLY LQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGDIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSRKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGSGGGGEVQLVESG LVQPGGSLRLSCAASGYTFTSYWMHWVRQAPGKGLEWIGEIKPSNGLTNYIEKFKNRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 251 Full size human GUCY2C
GenBank accession number NP_004954.2 MKTLLLDLALWSLLFQPGWLSFSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTYSWSTSYVYKNGTETEDCFWYLNA LEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYKVLLTYDTHVNKTYPVDMSPTFTW KNSKLPNDITGRGPQILMIAVFTLTGAVVLLLLVALLMLRKYRKDYELRQKKWSHIPPENIFPLETNETNHVSLKIDDDKRRDTIQRLRQCKYDKKRVILKDLKHNDGNFTEKQKIELNKLLQIDYYNLTKFYGTVKLDTMIFGVIEYCERGSLREVLNDTISYPDGTFMDWEFKISVLYDIAKGMSYLHSS KTEVHGRLKSTNCVVDSRMVVKITDFGCNSILPPKKDLWTAPEHLRQANISQKGDVYSYGIIAQEIILRKETFYTLSCRDRNEKIFRVENSNGMKPFRPDLFLETAEEKELEVYLLVKNCWEEDPEKRPDFKKIETTLAKIFGLFHDQKNESYMDTLIRRLQLYSRNLEHLVEERTQLYKAERDRADRLNFMLLPRLVVKSL KEKGFVEPELYEEVTIYFSDIVGFTTICKYSTPMEVVDMLNDIYKSFDHIVDHHDVYKVETIGDAYMVASGLPKRNGNRHAIDIAKMALEILSFMGTFELEHLPGLPIWIRIGVHSGPCAAGVVGIKMPRYCLFGDTVNTASRMESTGLPLRIHVSGSTIAILKRTECQFLYEVRGETYLKGRGNETTYWLTGMKDQKFNLPTPPTV ENQQRLQAEFSDMIANSLQKRQAAGIRSQKPRRVASYKKGTLEYLQLNTTDKESTYF 252 Full size human GUCY2C
GenBank accession number NM_004963.3 GACCAGAGAGAAGCGTGGGGAAGAGTGGGCTGAGGGACTCCACTAGAGGCTGTCCATCTGGATTCCCTGCCTCCCTAGGAGCCCAACAGAGCAAAGCAAGTGGGCACAAGGAGTATGGTTCTAACGTGATTGGGGTCATGAAGACGTTGCTGTTGGACTTGGCTTTGTGGTCACTGCTCTTCCAGCCCGGGTGGCTGTCCTTTAGTTCCCAGGTGAGTCAGAACTGCCACAATGGCAGCTATGAAA TCAGCGTCCTGATGATGGGCAACTCAGCCTTTGCAGAGCCCCTGAAAAACTTGGAAGATGCGGTGAATGAGGGGCTGGAAATAGTGAGAGGACGTCTGCAAAATGCTGGCCTAAATGTGACTGTGAACGCTACTTTCATGTATTCGGATGGTCTGATTCATAACTCAGGCGACTGCCGGAGTAGCACCTGTGAAGGCCTCGACCTACTCAGGAAAATTTCAAATGCACAACGGATGGGCTGTGTCCTCATAG CCCTCATGTACATACTCCACCTTCCAGATGTACCTTGACACAGAATTGAGCTACCCCATGATCTCAGCTGGAAGTTTTGGATTGTCATGTGACTATAAAGAAACCTTAACCAGGCTGATGTCTCCAGCTAGAAAGTTGATGTACTTCTTGGTTAACTTTTGGAAAACCAACGATCTGCCCTTCAAAACTTATTCCTGGAGCACTTCGTATGTTTACAAGAATGGTACAGAAACTGAGGACTGTTTCTGGTA CCTTAATGCTCTGGAGGCTAGCGTTTCCTATTTCTCCCACGAACTCGGCTTTAAGGTGGTGTTAAGACAAGATAAGGAGTTTCAGGATATCTTAATGGACCACAACAGGAAAAGCAATGTGATTATTATGTGTGGTGGTCCAGAGTTCCTCTACAAGCTGAAGGGTGACCGAGCAGTGGCTGAAGACATTGTCATTATTCTAGTGGATCTTTTCAATGACCAGTACTTTGAGGACAATGTCAC AGCCCCTGACTATATGAAAAATGTCCTTGTTCTGACGCTGTCTCCTGGGAATTCCCTTCTAAATAGCTCTTTCTCCAGGAATCTATCACCAACAAAACGAGACTTTGCTCTTGCCTATTTGAATGGAATCCTGCTCTTTGGACATATGCTGAAGATATTTCTTGAAAATGGAGAAAATATTACCACCCCCAAATTTGCTCATGCTTTCAGGAATCTCACTTTTGAAGGGTATGACGGTCCAGTGACCTTGGATGACT GGGGGGATGTTGACAGTACCATGGTGCTTCTGTATACCTCTGTGGACACCAAGAAATACAAGGTTCTTTTGACCTATGATACCCACGTAAATAAGACCTATCCTGTGGATATGAGCCCCACATTCACTTGGAAGAACTCTAAACTTCCTAATGATATTACAGGCCGGGGCCCTCAGATCCTGATGATTGCAGTCTTCACCTCACTGGAGCTGTGGTGCTGCTCCTGCTCGTCGCTCTCCTGATGC TCAGAAAATATAGAAAAGATTATGAACTTCGTCAGAAAAAATGGTCCCACATTCCTCCTGAAAATATCTTTCCTCTGGAGACCAATGAGACCAATCATGTTAGCCTCAAGATCGATGATGACAAAAGACGAGATACAATCCAGAGACTACGACAGTGCAAATACGACAAAAAGCGAGTGATTCTCAAAGATCTCAAGCACAATGATGGTAATTTCACTGAAAAACAGAAGATAGAATTGAACAAGTTGCTTCAGATTG ACTATTACAACCTGACCAAGTTCTACGGCACAGTGAAACTTGATACCATGATCTTCGGGGTGATAGAATACTGTGAGAGAGGATCCCTCCGGGAAGTTTTAAATGACACAATTTCCTACCCTGATGGCACATTCATGGATTGGGAGTTTAAGATCTCTGTCTTGTATGACATTGCTAAGGGAATGTCATATCTGCACTCCAGTAAGACAGAAGTCCATGGTCGTCTGAAATCTACCAACTGCGTAGTG GACAGTAGAATGGTGGTGAAGATCACTGATTTTGGCTGCAATTCCATTTTACCTCCAAAAAAGGACCTGTGGACAGCTCCAGAGCACCTCCGCCAAGCCAACATCTCTCAGAAAGGAGATGTGTACAGCTATGGGATCATCGCACAGGAGATCATCCTGCGGAAAGAAACCTTCTACACTTTGAGCTGTCGGGACCGGAATGAGAAGATTTTCAGAGTGGAAAATTCCAATGGAATGAAACCCTTCCGCCCA GATTTATTCTTGGAAACAGCAGAGGAAAAAAGAGCTAGAAGTGTACCTACTTGTAAAAAACTGTTGGGAGGAAGATCCAGAAAAGAGACCAGATTTCAAAAAAATTGAGACTACACTTGCCAAGATATTTGGACTTTTTCATGACCAAAAAAATGAAAGCTATATGGATACCTTGATCCGACGTCTACAGCTATATTCTCGAAACCTGGAACATCTGGTAGAGGAAAGGACACAGCTGTACAAGGCAGAGAGGGACA GGGCTGACAGACTTAACTTTATGTTGCTTCCAAGGCTAGTGGTAAAGTCTCTGAAGGAGAAAGGCTTTGTGGAGCCGGAACTATATGAGGAAGTTACAATCTACTTCAGTGACATTGTAGGTTTCACTACTATCTGCAAATACAGCACCCCCATGGAAGTGGTGGACATGCTTAATGACATCTATAAGAGTTTTGACCACATTGTTGATCATCATGATGTCTACAAGGTGGAAACCATCGGTGATGCGTACAT GGTGGCTAGTGGTTTGCCTAAGAGAAATGGCAATCGGCATGCAATAGACATTGCCAAGATGGCCTTGGAAATCCTCAGCTTCATGGGGACCTTTGAGCTGGAGCATCTTCCTGGCCTCCCAATATGGATTCGCATTGGAGTTCACTCTGGTCCCTGTGCTGCTGGAGTTGTGGGAATCAAGATGCCTCGTTATTGTCTATTTGGAGATACGGTCAACACAGCCTCTAGGATGGAATCCACTGGCCTCCC TTTGAGAATTCACGTGAGTGGCTCCACCATAGCCATCCTGAAGAGAACTGAGTGCCAGTTCCTTTATGAAGTGAGAGGAGAAACATACTTAAAGGGAAGAGGAAATGAGACTACCTACTGGCTGACTGGGATGAAGGACCAGAAATTCAACCTGCCAACCCCTCCTACTGTGGAGAATCAACAGCGTTTGCAAGCAGAATTTTCAGACATGATTGCCAACTCTTTACAGAAAAGACAGGCAGCAGGGATAAGAAG CCAAAAACCCAGACGGGTAGCCAGCTATAAAAAAGGCACTCTGGAATACTTGCAGCTGAATACCACAGACAAGGAGAGCACCTATTTTTAAACCTAAATGAGGTATAAGGACTCACACAAATTAAAATACAGCTGCACTGAGGCAGCGAACCTCAAGTGTCCTGAAAGCTTACATTTTCCTGAGACCTCAATGAAGCAGAAATGTACTTAGGCTTGGCTGCCCTGTCTGGAACATGGACTTTCTTGCATGAATCA GATGTGTGTTCTCAGTGAAATAACTACCTTCCACTCTGGAACCTTATTCCAGCAGTTGTTCCAGGGAGCTTCTACCTGGAAAAGAAAAGAAATGAATAGACTATCTAGAACTTGAGAAGATTTATTCTTATTTCATTTATTTTTTGTTTGTTTATTTTTATCGTTTTTGTTTACTGGCTTTCCTTCTGTATTCATAAGATTTTTTAAATTGTCATAATTATATTTTAAATACCCATCTTCATTAAAAGTATATT TAACTCATAATTTTTGCAGAAAATATGCTATATATTAGGCAAGAATAAAAGCTAAAGGTTTCCCAAAAAAAAAA 253 Full size GUCY2C cynomolgus macaque sqvsqnchngsyeisvlmmdnsafaeplenvedavnegleivrgrlqnaglnvtvnasfmysdglihnsgdcrsstcegldllrkisnakrmgcvlmgpsctystfqmyldtelsypmisagsfglscdyketltrlmsparkltyflvnfwktndlpfktyswstsyvykngtesedcfwylnaleasvsyf shelsfklvlrqdkefqdilmdhnrksnvivmcgdpeflyklkgdravaediviilvdlfndqyfednvtapdymknvlvltqspgnsllnssfsrnlsptkrdfalaylngillfghmlktflengenittpkfahafrnltfegydgpvtlddwgdvdstmvllytsvdtkkykvlltydthvnq tnpvdmsptftwknsklpnditdrgpqilmiavftltgavvllllvallmlrkykkdyelrqkkwshippenifpletnetnhvslkidddkrrdtiqrlrqckydkkrvilkdlkhndgnftekqkielnkllqidyynltkfygtvkldtmifgvieycergslrevlndtisypdgtfmdwef kisvlydiakgmsylhssktevhgrlkstncvvdsrmvvkitdfgcnsilppkkdlwtapehlrqanvsqkgdvysygiiaqeiilrketfytsscrdrnekifrvensngmkpfrpdlfletaeekelevyllvkscweedpekrpdfkkiettlakifglfhdqknesymdtlirrlqlysrnlehlveertqlyka erdradrlnfmllprlvvkslkekgfvepelyeevtiyfsdivgfttickystpmevvdmlndiyksfdhivdhhdvykvetigdaymvasglpkrngnrhaidiakmaleilsfmgtfelehlpglpiwirigvhsgpcaagvvgikmpryclfgdtvntasrmestglplrihvsgstiailkrtecqflyevrgety lkgrgnettywltgmkdqkfnlptpptvenqqrlqaefsdmianslqkrqaagirsqkprrvasykkgtleylqlnttdkestyf 254 Full size GUCY2C cynomolgus macaque tcacaggtgagtcagaactgccacaatggcagctatgaaatcagcgtcctgatgatggacaactcagcctttgcagagcccctggaaaacgtggaagatgcggtgaatgaggggctggaaatagtgagaggacgtctgcaaaacgctggcctaaatgtgactgtgaatgcttctttcatgtattcggatggtctgattcat aactccggcgactgccggagcagcacctgtgaaggccttgacctactcaggaaaatttcaaatgcaaaacggatgggctgtgtcctcatggggccctcatgtacatactccaccttccagatgtaccttgacacagaattgagctaccccatgatctcagctggaagttttggattgtcatgtgactataaagaaaccttaaccaggctgatgtct ccagctagaaagttgacatacttcttggttaacttttggaaaaccaatgatctacccttcaaaacttattcctggagcacttcgtatgtttacaagaatggtacggagtccgaggactgtttctggtaccttaacgctctggaggccagtgtttcctatttctcccacgaactcagttttaagttggtgttaagacaagataaggagtt tcaggatatcttaatggaccacaacaggaaaagcaatgtgattgttatgtgtggtgatccagagttcctctacaagttgaagggtgaccgagcagtggctgaagacattgtcattattctagtggatcttttcaatgaccagtactttgaggacaatgtcacagcccctgactatatgaaaaatgtccttgttctgacgcag tctcctggcaattccctcctaaatagctctttctccaggaatctatccccaacaaaacgagactttgctcttgcctatttgaatggaatcctgctctttggacatatgctaaagacatttcttgaaaatggagaaaatattaccacccccaaatttgctcatgcttttcaggaatctcacttttgaagggtatgacggtccagtgaccttggatg actggggggatgtggacagtaccatggtgcttctgtatacgtctgtggacaccaagaaatacaaggttcttttgacctatgatacccacgtaaatcagaccaaccctgtggatatgagccccacattcacttggaagaactctaaacttcctaatgatattacagaccggggccctcagatcctgatgattgcagtcttcaccctcaccggagctgtggt gctgctcctgcttgtcgctctcctgatgctcagaaaatataaaaaagattatgaacttcgtcagaaaaaatggtcccacattcctcctgaaaatatctttcctctggagaccaatgagaccaatcacgttagcctgaagatcgatgatgacaaaagacgagatacaatccagagactacgacagtgcaaatacgacaaaaagcgag tgattctcaaagatctcaagcacaatgatggtaatttcactgaaaaacagaagatagaattgaacaagttgcttcagattgactattacaacctgaccaagttctatggcaccgtgaaacttgataccatgatcttcggggtgatagaatactgtgagagaggatccctccgggaagttttaaatgacacaatttcctaccctgatggcacattcatggattgggagt ttaagatctctgtcctgtatgacattgctaagggaatgtcatatctgcactccagtaagacagaagtccatggtcgtctgaaatctaccaactgcgtagtggacagtagaatggtggtgaagatcactgattttggctgcaattccatttacctccaaaaaaagacctgtggacagctccagagcacctccgccaagccaacgtctctcagaaagg agatgtgtacagctacgggatcatcgcacaggagatcatcctgcggaaagaaaccttctacacttcgagctgtcgagaccggaacgagaagattttcagagtggaaaattccaatggaatgaaacccttccgtccagatttattcttggaaacggcagaggaaaaagagctagaagtgtacctacttgtaaaaagctgttgggaagaagat ccagaaaagagaccagatttcaaaaaaattgagactacacttgccaagatatttggactttttcatgaccaaaaaaatgaaagctatatggataccttgatccgacgtctacagctatattctcgaaacctggaacatctggtagaggaaaggacacagctatacaaggcagagagggacagggctgacagacttaactttatgttgcttccaaggctagtgg taaagtctctgaaggagaaaggctttgtagagccggaactatatgaggaagttacaatctacttcagtgacattgtaggtttcactactatctgcaaatacagcacccccatggaagtggtggacatgcttaatgacatctataagagttttgaccacattgttgatcatcatgatgtctacaaggtggaaaccattggtgatgcctacatggtggctag tggtttgcctaagagaaatggcaatcggcatgcaatagacattgccaagatggccttggaaatcctcagcttcatggggacctttgagctggagcatcttcctggcctcccaatatggattcgcattggcgttcactctggtccctgcgctgctggagttgtgggaatcaagatgcctcgttattgtctatttggagatacagtcaacacag cctctaggatggaatccactggcctccctttgaggattcatgtgagtggctccaccatagccattctgaagagaactgagtgccagttcctgtatgaagtgagaggagaaacgtacttaaagggaagaggaaatgagactacctactggctgaccgggatgaaggaccagaaattcaacctgccaacccctcctactgtggagaatcaacagcgtttgcaagcagaattt tcagacatgattgccaactctttacagaaaagacaggcggcagggataagaagccaaaaacccagacgagtagccagctataaaaaaggcactctggaatacttgcaactgaataccacggacaaggagagcacctatttt 255 Full size GUCY2C mouse
GenBank registration number NP_001120790.1 MTSLLGLAVRLLLFQPALMVFWASQVRQNCRNGSYEISVLMMDNSAYKEPMQNLREAVEEGLDIVRKRLREADLNVTVNATFIYSDGLIHKSGDCRSSTCEGLDLLREITRDHKMGCALMGPSCTYSTFQMYLDTELNYPMISAGSYGLSCDYKETLTRILPPARKLMYFLVDFWKVNNASFKPFSWNSSYVYKNGSEPEDCFWY LNALEAGVSYFSEVLNFKDVLRRSEQFQEILTGHNRKSNVIVMCGTPESFYDVKGDLQVAEDTVVILVDLFSNHYFEENTTAPEYMDNVLVLTLPSEQSTSNTSVAERFSSGRSDFSLAYLEGTLLFGHMLQTFLENGENVTGPKFARAFRNLTFQGFAGPVTLDDSGDIDNIMSLLYVSLDTRKYKVLMKYDTHKNKTIPVAENPNFIWKN HKLPNDVPGLGPQILMIAVFTLTGILVVLLLIALLVLRKYRRDHALRQKKWSHIPSENIFPLETNETNHISLKIDDDRRRDTIQRVRQCKYDKKKVILKDLKHSDGNFSEKQKIDLNKLLQSDYYNLTKFYGTVKLDTRIFGVVEYCERGSLREVLNDTISYPDGTFMDWEFKISVLNDIAKGMSYLHSSKIEVHG RLKSTNCVVDSRMVVKITDFGCNSILPPKKDLWTAPEHLRQATISQKGDVYSFAIIAQEIILRKETFYTLSCRDHNEKIFRVENSYGKPFRPDLFLETADEKELEVYLLVKSCWEEDPEKRPDFKKIESTLAKIFGLFHDQKNESYMDTLIRRLQLYSRNLEHLVEERTQLYKAERDRADHLNFMLLPRLVVKSLKEKGIVE PELYEEVTIYFSDIVGFTTICKYSTPMEVVDMLNDIYKSFDQIVDHHDVYKVETIGDAYVVASGLPMRNGNRHAVDISKMALDILSFIGTFELEHLPGLPVWIRIGVHSGPCAAGVVGIKMPRYCLFGDTVNTASRMESTGLPLRIHMSSSTITILKRTDCQFLYEVRGETYLKGRGTETTYWLTGMKDQEYNLPSPPTVENQQRLQTE FSDMIVSALQKRQASGKKSRRPTRVASYKKGFLEYMQLNNSDDHDSTYF 256 Full size GUCY2C mouse
GenBank registration number NM_001127318.1 GACCAGTGTGGCAAGACCAGAAAGGTGCGTGGGGAAGAAGACCAAGGGACTCTGCTAGCGACTCTCCAGAGGGGCTCCCTGTGTCTCTAAAAGCGAGCAATCCAGGGGCATGGTGCTACGGTGAGCCAGGTCATGACGTCACTGCTGGGCTTGGCTGTGCGGTTACTGCTCTTCCAGCCCGCGCTGATGGTGTTCTGGGCCTCTCAGGTGAGGCAGAACTGCCGCAATGGCAGCTACGAGATC AGCGTCCTGATGATGGACAACTCAGCCTACAAAGAACCTATGCAAAACCTGAGGGAGGCTGTGGAGGAAGGACTGGACATAGTGCGAAAGCGCCTGCGTGAAGCCGACCTAAATGTGACTGTGAACGCGACTTTCATCTACTCCGAACGGTCTGATTCATAAGTCAGGTGACTGCCGGAGCAGCACCTGTGAAGGCCTTGACCTACTCAGGGAGATTACAAGAGATCATAAGATGGGCTGCGCCCTCATGGGGCCCTCG TGCACGTATTCCACCTTCCAGATGTACCTCGACACAGAGTTGAACTATCCCATGATTTCCGCTGGAAGTTATGGATTGTCCTGTGACTATAAGGAAACCCTAACCAGGATCCTGCCTCCAGCCAGGAAGCTGATGTACTTCTTGGTCGATTTCTGGAAAGTCAACAATGCATCTTTCAAACCCTTTTCCTGGAACTCTTCGTATGTTTACAAGAATGGATCGGAACCTGAAGATTGTTTCTGGTACC TCAATGCTCTGGAGGCTGGGGTGTCCTATTTTTCTGAGGTGCTCAACTTCAAGGATGTACTGAGACGCAGCGAACAGTTCCAGGAAATCTTAACAGGCCATAACAGAAAGAGCAATGTGATTGTTATGTGTGGCACGCCAGAAAGCTTCTATGATGTGAAAGGTGACCTCCAAGTGGCTGAAGATACTGTTGTCATCCTGGTAGATCTGTTCAGTAACCATTACTTTGAGGAGAACACCACAGCTCC TGAGTATATGGACAATGTCCTCGTCCTGACGCTGCCGTCTGAACAGTCCACCTCAAACACCTCTGTCGCCGAGAGGTTTTCATCGGGGAGAAGTGACTTTTCTCTCGCTTACTTGGAGGGAACCTTGCTATTTGGACACATGCTGCAGACGTTTCTTGAAAATGGAGAAAATGTCACGGGTCCCAAGTTTGCTCGTGCATTCAGGAATCTCACTTTTCAAGGCTTTGCAGGACCTGTGACTCTGGATGACAG TGGGGACATTGACAACATTATGTCCCTTCTGTATGTGTCTCTGGATACCAGGAAATACAAGGTTCTTATGAAGTATGACACCCACAAAAACAAAACTATTCCGGTGGCTGAGAACCCCAACTTCATCTGGAAGAACCACAAGCTCCCCCAATGACGTTCCTGGGCTGGGCCCTCAAATCCTGATGATTGCCGTCTTCACGCTCACGGGGATCCTGGTAGTTCTGCTGCTGATTGCCCTCCTCGTGCTGA GAAAATACAGAAGAGATCATGCACTTCGACAGAAGAAATGGTCCCACATTCCTTCTGAAAACATCTTTCCTCTGGAGACCAACGAGACCAACCACATCAGCCTGAAGATTGACGATGACAGGAGACGAGACACAATCCAGAGAGTGCGACAGTGCAAATACGACAAGAAGAAAGTGATTCTGAAAGACCTCAAGCACAGCGACGGGAACTTCAGTGAGAAGCAGAAGATAGACCTGAACAAGTTGCTGCAGTCTG ACTACTACAACCTGACTAAGTTCTACGGCACCGTGAAGCTGGACACCAGGATCTTTGGGGTGGTTGAGTACTGCGAGAGGGGATCCCTCCGGGAAGTGTTAAACGACACAATTTCCTACCCTGACGGCACGTTCATGGATTGGGAGTTTAAGATCTCTGTCTTAAATGACATCGCTAAGGGGATGTCCTACCTGCACTCCAGTAAGATTGAAGTCCACGGGCGTCTCAAATCCACCAACTGCGTGGTG GACAGCCGCATGGTGGTGAAGATCACCGACTTTGGGTGCAATTCCATCCTGCCTCCAAAAAAAGACCTGTGGACGGCCCCGGAGCACCTGCGCCAGGCCACCATCTCTCAGAAAGGAGACGTGTACAGCTTCGCCATCATTGCCCAGGAGATCATCCTCCGTAAGGAGACTTTTTACACGCTGAGCTGTCGGGATCACAATGAGAAGATTTTCAGAGTGGAAAATTCATACGGGAAACCTTTCCGCCCAGACCTC TTCCTGGAGACTGCAGATGAGAAGGAGCTGGAGGTCTATCTACTTGTCAAAAGCTGTTGGGAGGAGGATCCAGAAAAGAGGCCAGATTTCAAGAAAATCGAGAGCACACTGGCCAAGATATTTGGCCTTTTCCATGACCAGAAAAACGAGTCTTACATGGACACCTTGATCCGACGTCTCCAGCTGTACTCTCGAAACCTGGAACATCTGGTGGAGGAAAGGACTCAGCTGTACAAGGCGGAGGGCAAGGGCTGA CCACCTTAACTTCATGCTCCTCCCACGGCTGGTGGTAAAGTCACTGAAGGAGAAAGGCATCGTGGAGCCAGAGCTGTACGAAGAAGTCACAATCTACTTCAGTGACATTGTGGGCTTCACCACCATCTGCAAGTATAGCACGCCCATGGAGGTGGTGGACATGCTCAACGACATCTACAAGAGCTTTGACCAGATTGTGGACCACCATGACGTCTACAAGGTAGAAACCATCGGTGACGCCTACG TGGTGGCCAGCGGTCTGCCTATGAGAAACGGCAACCGACACGCGGTAGACATTTCCAAGATGGCCTTGGACATCCTCAGCTTCATAGGGACCTTTGAGTTGGAGCATCTCCCTGGCCTCCCCGTGTGGATCCGCATTGGAGTTCATTCTGGGCCCTGCGCTGCTGGTGTTGTGGGGATCAAGATGCCTCGCTATTGCCTGTTTGGAGACACTGTCAACACTGCCTCCAGGATGGAATCCACCGGCCTCCCC TTGAGGATTCACATGAGCAGCTCCACCATAACCATCCTGAAGAGAACGGATTGCCAGTTCCTGTATGAAGTGAGGGGAGAAACCTACTTAAAGGGAAGAGGGACAGACCACATACTGGCTGACTGGGATGAAGGACCAAGAATACAACCTGCCATCCCCACCGACAGTGGAGAACCAACAGCGTCTGCAGACTGAGTTCTCAGACATGATCGTTAGCGCCTTACAGAAAAGACAGGCCTCGGGCAAGAAG AGCCGGAGGCCCACTCGGGTGGCCAGCTACAAGAAAGGCTTTCTGGAATACATGCAGCTGAACAATTCAGACCACGATAGCACCTATTTTTAGACCAAGTGAGGTCTGAGAACTGACAGTAGCAACCTCCTATATCATGAATCTGTATTTTCCAGAGACCTCAACAACATAGACAAGCACTTAGCCTCAGTGCCCTGACTGGAACGTAGAACCAACCCCTCAAGTCATGTGGGTCTGATTTTGGGTTGGTT GGTTGGTTGGTTGGTTGGTTTTGGTTTTGTTGAGACAGAGTTTCGTGTATCCCAAGCTGGTCTCAAACTCACTGTGTAGCAGCAGATGACTTTGGACTTCTAAGATCATCCGTGTGTTCCTGACAGTGTGATGAGTGTATACGTCAGAGCCCTGTCCCACAGTTCTCCATGGAGCATCAACCTGAATGGAGGAGCGAGGAGGGGGAATGTCCTGTGCTGTACAGAACTTGGGGTTTTGTTCT AATTTTATCTCTGGTTTGGTTTTGTTTTCTGGCTCCTTCCCTCTCTATGTGTGAGAGAAGTTTTTTAAATTGTCTGAATTGTATGCTAAGTAGCTTATCTACAAGAAAGTGTGTTTAACTAGTGATTTTTGCAGAAACCATGCTGGATATTAGGTAAAAAATAAAAGTGTTTAGAGTCTAAAAAAAAAAAAAAAA 257 GUCY2C-0074_CDR1 VH TYWMQ 258 GUCY2C-0077_CDR1 VH KYWMQ 259 GUCY2C-0098_CDR1 VH SYWMH 260 GUCY2C-0104_CDR1 VH DTYIH 261 GUCY2C-0105_CDR1 VH DYIML 262 GUCY2C-0074_CDR2 VH YPGDGM 263 GUCY2C-0077_CDR2 VH YPGDGF 264 GUCY2C-0098_CDR2 VH KPSNGL 265 GUCY2C-0104_CDR2 VH DPANGN 266 GUCY2C-0105_CDR2 VH NPYYGS 267 GUCY2C-1554_VH; GUCY2C-1608_CDR2 VH KPSNEL 268 CD3-0006_VH,
2B5-1038 VH,
2B5-1039 VH,
2B5-1040 VH,
CDR1 (Kabat) DYYMT 269 CD3-0001_CDR2 VH RNRARGYT 270 CD3-0006_VH,
2B5-1038 VH,
2B5-1039 VH,
2B5-1040 VH,
CDR2 (Chothia) RNQARGYT 271 CDR1 VH huIGHV3-7 SYWMS 272 CDR2 VH huIGHV3-7 KQDGSE 273 2B5-1038 VH,
2B5-1039 VH,
2B5-1040 VH EVQLVESGGGLVQPGGSLRLSCAAS GFTFS DY YMT WVRQAPGKGLEWVA FI RNQARGYT SDHNPSVKG RFTISRDNAKNSLYLQMNSLRAEDTAVYYCAR DRPSYYVLDY WGQGTTVTVSS 274 VL 2B5-1038 DIQMTQSPSSLSSASVGDRVTITC TSDQSLFNVRSGKNYLA WYQQKPGKAPKLLIY WASDRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK 275 VL 2B5-1039 DIQMTQSPSSLSSASVGDRVTITC TSSESLFNVRSGKNYLA WYQQKPGKAPKLLIY WASDRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK 276 VL 2B5-1040 DIQMTQSPSSLSSASVGDRVTITC TSSQSLFNVRSGKNYLA WYQQKPGKAPKLLIY WASDRES GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC KQSYDLFT FGGGTKVEIK 277 VH 2B5-1038,
VH 2B5-1039,
VH 2B5-1040,
CDR1 (Chothia) GFTFSDY 278 CDR1 VL 2B5-1038 TSDQSLFNVRSGKNYLA 279 CDR1 VL 2B5-1039 TSSESLFNVRSGKNYLA 280 CDR1 VL 2B5-1040 TSSQSLFNVRSGKNYLA 281 VL 2B5-1038,
VL 2B5-1039,
VL 2B5-1040,
CDR2 WASDRES 282 GUCY2C-1678_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 283 GUCY2C-1678_cavity DIQMTQSPSSLSASVGDRVTITCTSDQSLFNVRSGKNYLAWYQQKPGKAPKLLIYWASDRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 284 GUCY2C-1679_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 285 GUCY2C-1679_cavity DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSGKNYLAWYQQKPGKAPKLLIYWASDRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 286 GUCY2C-1680_protrusion DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSREEMTKNQVSLWCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 287 GUCY2C-1680_cavity DIQMTQSPSSLSASVGDRVTITCTSSESLFNVRSGKNYLAWYQQKPGKAPKLLIYWASDRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAY LQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCREEMTKNQVSLSCAVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 288 2B5-1038_VL GACATCCAGATGACCCAGTCCCCCTCTTCTCTGTCTGCCTCTGTGGGCGACAGAGTGACCATCACCTGCACAAGCGACCAGTCACTGTTTAATGTCCGCAGCGGCAAAAACTATCTTGCGTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGCTGCTGATCTACTGGGGCCAGTGACCGAATCCGGCGTGCCTTCCAGATTCTCCGGCTCTGGCTCTGGCACCGATTTCACCCTGACCATCT CCTCCCTCCAGCCTGAGGATTTCGCCACCTACTGCAAACAGTCTTACGACCTTTTTCACTTTTGGCGGCGGAACAAAGGTGGAGATCaag 289 2B5-1038_VH GAAGTGCAGCTTGTTGAATCTGGCGGCGGTTTGGTTCAGCCCGGTGGATCACTGCGACTCAGTTGCGCAGCTAGCGGCTTCACCTTTTCTGATTACTACATGACATGGGTACGACAGGCGCCAGGCAAGGGTTTGGAATGGGTAGCATTCATACGCAATCAGGCACGCGGGTACACTTCAGACCACAATCCCTCAGTAAAAGGAAGATTTACCATCTCAAGAGACAATGCCAAAAATTCACTCTAC CTGCAAATGAACTCACTTCGCGCCGAGGATACCGCCGTGTATTACTGTGCCAGAGACAGACCATCTTATTACGTGCTGGACTATTGGGGACAGGGCACTACAGTCACCGTCAGCTCT 290 2B5-1039_VL GACATCCAGATGACCCAGTCCCCCTCTTCTCTGTCTGCCTCTGTGGGCGACAGAGTGACCATCACCTGCACAAGCTCAGAGTCACTGTTTAATGTCCGCAGCGGCAAAAACTATCTTGCGTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGCTGCTGATCTACTGGGCCAGTGACCGAATCCGGCGTGCCTTCCAGATTCTCCGGCTCTGGCTCTGGCACCGATTTCACCCTGACCATCT CCTCCCTCCAGCCTGAGGATTTCGCCACCTACTGCAAACAGTCTTACGACCTTTTCACTTTTGGCGGCGGAACAAAGGTGGAGATCAAG 291 2B5-1039_VH GAAGTGCAGCTTGTTGAATCTGGCGGCGGTTTGGTTCAGCCCGGTGGATCACTGCGACTCAGTTGCGCAGCTAGCGGCTTCACCTTTTCTGATTACTACATGACATGGGTACGACAGGCGCCAGGCAAGGGTTTGGAATGGGTAGCATTCATACGCAATCAGGCACGCGGGTACACTTCAGACCACAATCCCTCAGTAAAAGGAAGATTTACCATCTCAAGAGACAATGCCAAAAATTCACTCTAC CTGCAAATGAACTCACTTCGCGCCGAGGATACCGCCGTGTATTACTGTGCCAGAGACAGACCATCTTATTACGTGCTGGACTATTGGGGACAGGGCACTACAGTCACCGTCAGCTCT 292 2B5-1040_VL GACATCCAGATGACCCAGTCCCCCTCTTCTCTGTCTGCCTCTGTGGGCGACAGAGTGACCATCACCTGCACAAGCTCACAGTCACTGTTTAATGTCCGCAGCGGCAAAAACTATCTTGCGTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGCTGCTGATCTACTGGGGCCAGTGACCGAATCCGGCGTGCCTTCCAGATTCTCCGGCTCTGGCTCTGGCACCGATTTCACCCTGACCATCT CCTCCCTCCAGCCTGAGGATTTCGCCACCTACTGCAAACAGTCTTACGACCTTTTCACTTTTGGCGGCGGAACAAAGGTGGAGATCAAG 293 2B5-1040_VH GAAGTGCAGCTTGTTGAATCTGGCGGCGGTTTGGTTCAGCCCGGTGGATCACTGCGACTCAGTTGCGCAGCTAGCGGCTTCACCTTTTCTGATTACTACATGACATGGGTACGACAGGCGCCAGGCAAGGGTTTGGAATGGGTAGCATTCATACGCAATCAGGCACGCGGGTACACTTCAGACCACAATCCCTCAGTAAAAGGAAGATTTACCATCTCAAGAGACAATGCCAAAAATTCACTCTAC CTGCAAATGAACTCACTTCGCGCCGAGGATACCGCCGTGTATTACTGTGCCAGAGACAGACCATCTTATTACGTGCTGGACTATTGGGGACAGGGCACTACAGTCACCGTCAGCTCT 294 GUCY2C-1696_VH EVQLVESGGGLVQPGGSLRLSCAASGTFSTRWMHWVRQAPGKGLEWIGEIKPSNKLTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARTITTTEGYWFLSDWGQGTLVTVSS 295 GUCY2C-1696_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGHSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 296 GUCY2C-1701_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTPLGYWFFDVWGQGTLVTVSS 297 GUCY2C-1701_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 298 GUCY2C-1702_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTPLGYWFFDVWGQGTLVTVSS 299 GUCY2C-1702_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 300 GUCY2C-1703_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNRWNNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 301 GUCY2C-1703_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 302 GUCY2C-1705_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSRGFTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 303 GUCY2C-1705_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 304 GUCY2C-1706_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSTWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS 305 GUCY2C-1706_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 306 GUCY2C-1708_VH EVQLVESGGGLVQPGGSLRLSCAASGTFFSRPWMHWVRQAPGKGLEWIGEIKPSTGWTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTTGYWFFDVWGQGTLVTVSS 307 GUCY2C-1708_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYKHSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 308 GUCY2C-1710_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSRGWTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARTITTTEGYWFFDVWGQGTLVTVSS 309 GUCY2C-1710_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYRHSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 310 GUCY2C-1713_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSHTWMHWVRQAPGKGLEWIGEIKPSRGFTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTQGYWFFDVWGQGTLVTVSS 311 GUCY2C-1713_VL DIQLTQSPSSSLSASVGDRVTITCRASESVNWYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 312 GUCY2C-1714_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSHTWMHWVRQAPGKGLEWIGEIKPSTKYTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARTIFTREGYWFFDVWGQGTLVTVSS 313 GUCY2C-1714_VL DIQLTQSPSSSLSASVGDRVTITCRASESVNWYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 314 GUCY2C-1715_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSTKYTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTIFTNEGYWFFDVWGQGTLVTVSS 315 GUCY2C-1715_VL DIQLTQSPSSLSSASVGDRVTITCRASESVSIYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK 316 Peptide 1 GUCY2C_ECD MKTLLLDLALWSLLFQPGWL 317 Peptide 2 GUCY2C_ECD LDLALWSLLFQPGWLSFSSQ 318 Peptide 3 GUCY2C_ECD WSLLFQPGWLSFSSQVSQNC 319 Peptide 4 GUCY2C_ECD QPGWLSFSSQVSQNCHNGSY 320 Peptide 5 GUCY2C_ECD SFSSQVSQNCHNGSYEISVL 321 Peptide 6 GUCY2C_ECD VSQNCHNGSYEISVLMMGNS 322 Peptide 7 GUCY2C_ECD HNGSYEISVLMMGNSAFAEP 323 Peptide 8 GUCY2C_ECD EISVLMMGNSAFAEPLKNLE 324 Peptide 9 GUCY2C_ECD MMGNSAFAEPLKNLEDAVNE 325 Peptide 10 GUCY2C_ECD AFAEPLKNLEDAVNEGLEIV 326 Peptide 11 GUCY2C_ECD LKNLEDAVNEGLEIVRGRLQ 327 Peptide 12 GUCY2C_ECD DAVNEGLEIVRGRLQNAGLN 328 Peptide 13 GUCY2C_ECD GLEIVRGRLQNAGLNVTVNA 329 Peptide 14 GUCY2C_ECD RGRLQNAGLNVTVNATFMYS 330 Peptide 15 GUCY2C_ECD NAGLNVTVNATFMYSDGLIH 331 Peptide 16 GUCY2C_ECD VTVNATFMYSDGLIHNSGDC 332 Peptide 17 GUCY2C_ECD TFMYSDGLIHNSGDCRSSTC 333 Peptide 18 GUCY2C_ECD DGLIHNSGDCRSSTCEGLDL 334 Peptide 19 GUCY2C_ECD NSGDCRSSTCEGLDLLRKIS 335 Peptide 20 GUCY2C_ECD RSSTCEGLDLLRKISNAQRM 336 Peptide 21 GUCY2C_ECD EGLDLLRKISNAQRMGCVLI 337 Peptide 22 GUCY2C_ECD LRKISNAQRMGCVLIGPSCT 338 Peptide 23 GUCY2C_ECD NAQRMGCVLIGPSCTYSTFQ 339 Peptide 24 GUCY2C_ECD GCVLIGPSCTYSTFQMYLDT 340 Peptide 25 GUCY2C_ECD GPSCTYSTFQMYLDTELSYP 341 Peptide 26 GUCY2C_ECD YSTFQMYLDTELSYPMISAG 342 Peptide 27 GUCY2C_ECD MYLDTELSYPMISAGSFGLS 343 Peptide 28 GUCY2C_ECD ELSYPMISAGSFGLSCDYKE 344 Peptide 29 GUCY2C_ECD MISAGSFGLSCDYKETLTRL 345 Peptide 30 GUCY2C_ECD SFGLSCDYKETLTRLMSPAR 346 Peptide 31 GUCY2C_ECD CDYKETLTRLMSPARKLMYF 347 Peptide 32 GUCY2C_ECD TLTRLMSPARKLMYFLVNFW 348 Peptide 33 GUCY2C_ECD MSPARKLMYFLVNFWKTNDL 349 Peptide 34 GUCY2C_ECD KLMYFLVNFWKTNDLPFKTY 350 Peptide 35 GUCY2C_ECD LVNFWKTNDLPFKTYSWSTS 351 Peptide 36 GUCY2C_ECD KTNDLPFKTYSWSTSYVYKN 352 Peptide 37 GUCY2C_ECD PFKTYSWSTSYVYKNGTETE 353 Peptide 38 GUCY2C_ECD SWSTSYVYKNGTETEDCFWY 354 Peptide 39 GUCY2C_ECD YVYKNGTETEDCFWYLNALE 355 Peptide 40 GUCY2C_ECD GTETEDCFWYLNALEASVSY 356 Peptide 41 GUCY2C_ECD DCFWYLNALEASVSYFSHEL 357 Peptide 42 GUCY2C_ECD LNALEASVSYFSHELGFKVV 358 Peptide 43 GUCY2C_ECD ASVSYFSHELGFKVVLRQDK 359 Peptide 44 GUCY2C_ECD FSHELGFKVVLRQDKEFQDI 360 Peptide 45 GUCY2C_ECD GFKVVLRQDKEFQDILMDHN 361 Peptide 46 GUCY2C_ECD LRQDKEFQDILMDHNRKSNV 362 Peptide 47 GUCY2C_ECD EFQDILMDHNRKSNVIMCG 363 Peptide 48 GUCY2C_ECD LMDHNRKSNVIIMCGGPEFL 364 Peptide 49 GUCY2C_ECD RKSNVIIMCGGPEFLYKLKG 365 Peptide 50 GUCY2C_ECD IIMCGGPEFLYKLKGDRAVA 366 Peptide 51 GUCY2C_ECD GPEFLYKLKGDRAVAEDIVI 367 Peptide 52 GUCY2C_ECD YKLKGDRAVAEDIVIILVDL 368 Peptide 53 GUCY2C_ECD DRAVAEDIVIILVDLFNDQY 369 Peptide 54 GUCY2C_ECD EDIVIIILVDLFNDQYLEDNV 370 Peptide 55 GUCY2C_ECD ILVDLFNDQYLEDNVTAPDY 371 Peptide 56 GUCY2C_ECD FNDQYLEDNVTAPDYMKNVL 372 Peptide 57 GUCY2C_ECD LEDNVTAPDYMKNVLVLTLS 373 Peptide 58 GUCY2C_ECD TAPDYMKNVLVLTLSPGNSL 374 Peptide 59 GUCY2C_ECD MKNVLVLTLSPGNSLLNSSF 375 Peptide 60 GUCY2C_ECD MKNVLVLTLSPGNSLLNSSF 376 Peptide 61 GUCY2C_ECD VLTLSPGNSLLNSSFSRNLS 377 Peptide 62 GUCY2C_ECD PGNSLLNSSFSRNLPTKRD 378 Peptide 63 GUCY2C_ECD LNSSFSRNLSPKRDFALAY 379 Peptide 64 GUCY2C_ECD SRNLSPTKRDFALAYLNGIL 380 Peptide 65 GUCY2C_ECD PTKRDFALAYLNGILLFGHM 381 Peptide 66 GUCY2C_ECD FALAYLNGILLFGHMLKIFL 382 Peptide 67 GUCY2C_ECD LNGILLFGHMLKIFLENGEN 383 Peptide 68 GUCY2C_ECD LFGHMLKIFLENGENITTPK 384 Peptide 69 GUCY2C_ECD LKIFLENGENITTPKFAHAF 385 Peptide 70 GUCY2C_ECD ENGENITTPKFAHAFRNLTF 386 Peptide 71 GUCY2C_ECD ITTPKFAHAFRNLTFEGYDG 387 Peptide 72 GUCY2C_ECD FAHAFRNLTFEGYDGPVTLD 388 Peptide 73 GUCY2C_ECD RNLTFEGYDGPVTLDDWGDV 389 Peptide 74 GUCY2C_ECD EGYDGPVTLDDWGDVDSTMV 390 Peptide 75 GUCY2C_ECD PVTLDDWGDVDSTMVLLYTS 391 Peptide 76 GUCY2C_ECD DWGDVDSTMVLLYTSVDTKK 392 Peptide 77 GUCY2C_ECD DSTMVLLYTSVDTKKYKVLL 393 Peptide 78 GUCY2C_ECD LLYTSVDTKKYKVLLTYDTH 394 Peptide 79 GUCY2C_ECD VDTKKYKVLLTYDTHVNKTY 395 Peptide 80 GUCY2C_ECD YKVLLTYDTHVNKTYPVDMS 396 Peptide 81 GUCY2C_ECD TYDTHVNKTYPVDMSPTFTW 397 Peptide 82 GUCY2C_ECD VNKTYPVDMSPTFTWKNSKL 398 Peptide 83 GUCY2C_ECD PVDMSPTFTWKNSKLPNDIT 399 Peptide 84 GUCY2C_ECD PTFTWKNSKLPNDITGRGPQ 400 Peptide 85 GUCY2C_ECD KNSKLPNDITGRGPQILMIA 401 Peptide 86 GUCY2C_ECD PNDITGRGPQILMIAVFTLT 402 Peptide 87 GUCY2C_ECD LRKISNAQRMGCVLIGPSCT 403 Peptide 88 GUCY2C_ECD NAQRMGCVLIGPSCTYSTFQ 404 Peptide 89 GUCY2C_ECD GCVLIGPSCTYSTFQMYLDT 405 Peptide GUCY2C_ECD NSGDCRSSTCEGLDLLRKISNAQRM 406 Peptide GUCY2C_ECD RSSTCEGLDLLRKIS 407 GUCY2C-1637_VH DIQMTQSPSSLSASVGDRVTITCTSSQSLFNVRSQKNYLAWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQSYDLFTFGGGTKVEIKGGGGSGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNS AYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSSGGCGGHHHHHH 408 GUCY2C-1637_VL DIQLTQSPSSLSASVGDRVTITCRASESVDYYGSSLLQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIKGGGSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMTWVRQAPGKGLEWVAFIRNQARGYTSDHNPSVKGRFTISRDNAKNS LYLQMNSLRAEDTAVYYCARDRPSYYVLDYWGQGTTVTVSSGGCGGDYKDDDDK 409 GUCY2C-1608_scFv evqlvesggglvqpggslrlscaasgftfssywmhwvrqapgkglewigeikpsneltnvhekfkdrftisvdkaknsaylqmnslraedtavyyctrtitttegywffdvwgqgtlvtvssggggsggggsggggsgdiqltqspsslsasvgdrvtitcrasesvdyygssllqwyqqk pgkapklliyaasklasgvpsrfsgsgsgtdftltisslqpedfatyycqqtrkaytfgqgtkleiktgsenlyfq 410 CID1814 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 411 CID1815 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVRQKCHNGTYEISVLMMDNSAYKEPLQNLRDAVEEGLDIVRKRLREAELNVTVNATFIYSDGLIHKSGDCRSSTCEGLDLLREITRDRKMGCVLMGPSCTYSTFQMYLDTELNYPMISAGSFGLSCDYKETLTRILPPARKLMYFLVDFWKVNNAPFK TFSWNSSYVYKNGSEPEDCFWYLNALEAGVSYFSEVLSFKDVLRRSEQFQEILMGRNRKSNVIVMCGTPETFYNVKGDLKVADDTVVILVDLFSNHYFEDDTRAPEYMDNVLVLTLPPEKFIANASVSGRFPSERSDFSLAYLEGTLLFGHMLQTFLENGESVTTPKFARAFRNLTFQGLEGPVTLDDSGDIDNIMCLLYVSLDTRKYKVLMAY DTHKNQTIPVATSPNFIWKNHRLPNDVPGLGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIMSSSYTSADTP SETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSSLRTLTPSSVGTSTSLTT TTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 412 CID1818 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSSRCNNNNYMINVMLMPDSDFPSTSENLTSAVEEALSTIQNELETEGVKVTVNASFHHFRSSLYVSQGCRTSTCEGVELIKQIFENGTLGCAVIGPACTYATYQMVSVETIPSLPLISVGSFGLSCDYKENLTRLLTPARKVNDFFYYFWNEIQQPFKTSTW ESVYLYKKTDNSEQCLWYMNALDAGVTQFSEKLKFKDIVRTQDQFRRLVKNPKRKSNVIIMCGTPADIRQDLGTETVDKDIVIILIDLFKNTYFRNTTSARYMQNVLVLTLPPANNNFSTRTTDTSLLEDDFVIGYYNAVLLFGHILKKFIFSQSPVLPTSFINEFRNITFEGAQGPVTLDEFGDIDNNLTLLYTTQSASDP QYRVLMYFNTQENDTYVVSTSPDFIWKSHRLPSDIPSTGPHRTSDQASGAHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIM SSSYTSADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSAVTPTPVTPSSLSTDIPTSLRTLTPS GTSTSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLALALAPLVLLSLL 413 CID1868 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEINVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYFDGLLHNSGDCRSSTCEGVDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFK TYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 414 CID1869 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGDSDFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGGCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 415 CID1870 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMVLDTELSYPMISAGSFGLSCDYKENLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 416 CID1871 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY TWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITSRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 417 CID1872 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFRDILMDHNRKSNVIIMCGGPAFLYKLKGTRAVAEDIVIILVDLFNDQYFRDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYK VLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIMSSSYTSAD TPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTSTSL TTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 418 CID1873 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHEFRNITFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 419 CID1874 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRTSTCEGVELIKQIFENGTLGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFK TYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 420 CID1875 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWNEIQQPFK TSTWESSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 421 CID1876 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKKTDNSEQCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTK KYKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIMSS SYTSADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVG TSTSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLALALAPLVLLSLL 422 CID1877 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELKFKDIVRTQKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 423 CID1878 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDDQFRRLVKNPKRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 424 CID1879 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGTETVDKDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 425 CID1880 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFRNTTSARYMQNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKYK VLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIMSSSYTSAD TPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTSTSL TTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 426 CID1881 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSASDPQYR VLMYFNTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 427 CID1939 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSSRCNNNNYMINVMLMPDSDFPSTSENLTSAVEEALSTIQNELETEGVKVTVNASFHHFRSSLYVSQGCRSSTCEGLDLLRKISNAQRMGCAVIGPACTYATYQMVSVETIPSLPLISVGSFGLSCDYKENLTRLLTPARKVNDFFYYFWNEIQQPFKTSTWES VYLYKKTDNSEQCLWYMNALDAGVTQFSEKLKFKDIVRTQDQFRRLVKNPKRKSNVIIMCGTPADIRQDLGTETVDKDIVIILIDLFKNTYFRNTTSARYMQNVLVLTLPPANNNFSTRTTDTSLLEDDFVIGYYNAVLLFGHILKKFIFSQSPVLPTSFINEFRNITFEGAQGPVTLDEFGDIDNNLTLLYTTQSASDPQ YRVLMYFNTQENDTYVVSTSPDFIWKSHRLPSDIPSTGPHRTSDQASGAHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIMSS SYTSADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVG TSTSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLALALAPLVLLSLL 428 CID1940 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSSRCNNNNYMINVMLMPDSDFPSTSENLTSAVEEALSTIQNELETEGVKVTVNASFHHFRSSLYVSQGCRSSTCEGLDLLRKISENGTLGCAVIGPACTYATYQMVSVETIPSLPLISVGSFGLSCDYKENLTRLLTPARKVNDFFYYFWNEIQQPFKTSTWES VYLYKKTDNSEQCLWYMNALDAGVTQFSEKLKFKDIVRTQDQFRRLVKNPKRKSNVIIMCGTPADIRQDLGTETVDKDIVIILIDLFKNTYFRNTTSARYMQNVLVLTLPPANNNFSTRTTDTSLLEDDFVIGYYNAVLLFGHILKKFIFSQSPVLPTSFINEFRNITFEGAQGPVTLDEFGDIDNNLTLLYTTQSASDPQ YRVLMYFNTQENDTYVVSTSPDFIWKSHRLPSDIPSTGPHRTSDQASGAHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSIMSS SYTSADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVG TSTSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLALALAPLVLLSLL 429 CID1941 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEINVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 430 CID1942 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYFDGLIHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 431 CID1943 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLLHNSGDCRSSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 432 CID1944 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRTSTCEGLDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 433 CID1945 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGVDLLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFK TYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 434 CID1946 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLELLRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 435 CID1947 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLIRKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 436 CID1948 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLKKISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFK TYSWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKK YKVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSY TSADTPSETSVYTSSETPSSSSPTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTSLRTLTPSSVGTS TSLTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 437 CID1949 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRQISNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 438 CID1950 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSSQVSQNCHNGSYEISVLMMGNSAFAEPLKNLEDAVNEGLEIVRGRLQNAGLNVTVNATFMYSDGLIHNSGDCRSSTCEGLDLLRKIFNAQRMGCVLIGPSCTYSTFQMYLDTELSYPMISAGSFGLSCDYKETLTRLMSPARKLMYFLVNFWKTNDLPFKTY SWSTSYVYKNGTETEDCFWYLNALEASVSYFSHELGFKVVLRQDKEFQDILMDHNRKSNVIIMCGGPEFLYKLKGDRAVAEDIVIILVDLFNDQYFEDNVTAPDYMKNVLVLTLSPGNSLLNSSFSRNLSPTKRDFALAYLNGILLFGHMLKIFLENGENITTPKFAHAFRNLTFEGYDGPVTLDDWGDVDSTMVLLYTSVDTKKY KVLLTYDTHVNKTYPVDMSPTFTWKNSKLPNDITGRGPQRTSDQASGAHHHHHHGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 439 CID1431 MQLLRCFSIFSVIASVLAQEGKPIPNPLLGLDSTGGGSGGGAQAAGAHSRTSDQASGAGAYPYDVPDYAGHGTSGGGGSGGTSTPSYTTSIISTETPSHSTPSSSTSITTTETPSHSTPSYTSSVSTSETTSHSTPSETSSSRTTESTSYSSPSSTSSNTITETSSHSTPSTATSISSTETPSSSIPSVSSSITVTESSSHSTPGATSTLTSSETSTWSTPSSTSSSIMSSYTS ADTPSETSVYTSSETPSSSSPTSTSLISSSKSTSTSTPSFTSSITSTETSSYSASSYTPSVSSTASSSKNTTSSTASISSAETVSSSSSSVSSTIPSSQSTSYSTPSFSSSATSSVTPLHSTPSLPSWVTTSKTTSHITPGLTSSMSSSETYSHSTPGFTSSITSTESTSESTPSLSSSTIYSTVSTSTTAITSHFTTSETAVTPTPVTPSSLSTDIPTTSLRTLTPSSVGTSTS LTTTTDFPSIPTDISTLPTRTHIISSSPSIQSTETSSLVGTTSPTMSTVRMTLRITENTPISSFSTSIVVIPETPTQTPPVLTSATGTQTSPAPTTVTFGSTDSSTSTLHKLQGDSGETIGKYIGAADSLGGSVLLLALAPLVLLSLL 440 GUCY2C-0118_
Rabbit IgG light chain clone 9H3 DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIKGDPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTSTQYNSHKEYTCRVT QGTTSVVQSFNRGDC 441 GUCY2C-0118_
Rabbit IgG heavy chain version of clone 9H3 QVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTVDKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSVRQSVGLYSLSSVV TSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPPREELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAV LDSDGSYFLYSKLSVPTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK 442 GUCY2C-0117_
Mouse IgG1 light chain clone 9H3 DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIKRTDAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTC EATHKTSSTSPIVKSFNRNEC 443 GUCY2C-0117_
Mouse IgG1 heavy chain clone 9H3 QVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTVDKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSST WPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQ PAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK 444 GUCY2C-0119_
Light chain_
Chimeric 9H3 human IgG1 3m in pTT5 DIVLTQSPASLAVSLGQRATISCRASESVDYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAMYFCQQTRKVYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK VYACEVTHQGLSSPVTKSFNRGEC 445 GUCY2C-0119_
Heavy chain
Chimeric 9H3 human IgG1 3m in pTT5 QVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEIKPSNGLTNYIEKFKNKATLTVDKSATTAYMQLSSLTAEDSAVYYCTRTITTTEGYWFFDVWGAGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

The antibody sequences of the present invention are numbered using Abysis AbM. Using the Abysis numbering system, AbM is assigned Kabat numbering and defines the beginning and end of each CDR according to Kabat rules, with the exception of CDR H1. CDR H1 is determined by AbM.

Example 27: Three Anti-CD3 Antibody Variants Showed Improved Affinity and Cytotoxicity in Anti-GUCY2C-CD3 Bispecific Antibodies

Three anti-CD3 antibody variants, 2B5-1038 (SEQ ID NOs: 273 and 274), 2B5-1039 (SEQ ID NOs: 273 and 275), and 2B5-1040 (SEQ ID NOs: 273 and 276), derived from 2B5v6, reformatted into bispecific diabody-Fc molecules paired with an anti-GUCY2c anti-tumor antibody sequence in one of the configurations shown in Figure 1, using standard cloning, expression and purification methods presented herein above. The resulting bispecific antibodies, shown in Table 39, were tested for in vitro cytotoxicity using a T cell redirection assay, binding affinity using a surface plasmon resonance (SPR) assay, and nonspecificity using AC-SIN. Immunogenicity risk was assessed using the Epivax Tool for in silico prediction of potential T cell epitopes. Intact mass spectroscopy data indicate that all bispecific antibodies are correctly paired and are 100% heterodimers. Table 39 shows that all three bispecific antibodies (1) are approximately 2-fold more active in an in vitro cytotoxicity assay compared to the control bispecific GUCY2C-1608 paired with anti-CD3 antibody 2B5v6 (Figure 18); (2) have a binding affinity for recombinant human CD3 as determined by SPR consistent with cytotoxicity activity; (3) have immunogenicity scores that are improved compared to controls; and (4) AC-SIN scores remained the same as those of the control bispecific antibody GUCY2C-1608.

Table 39

Bispecific antibody Anti-CD3 antibody variant Epivax scores (adjusted to tReg) Mass spectrometry (intact data) AC-SIN points Cytotoxicity in vitro (nM) KD (nM, SPR) GUCY2C-1608 2B5-0542 -33.7 100% heterodimer 3 0.772 127.88±1.63 GUCY2C-1678 2B5-1038 -52.61 100% heterodimer 2 0.247 63.55±0.49 GUCY2C-1679 2B5-1040 -47.58 100% heterodimer 3 0.194 72.02±0.36 GUCY2C-1680 2B5-1039 -53.51 100% heterodimer 3 0.385 78.57±0.9

Example 28: Affinity Maturation of Anti-GUCY2c Antibody Binding Domain

A phage display approach was used to increase the affinity of the anti-GUCY2c antibody binding domain for GUCY2c. Four phage display libraries were constructed and prepared using the methods described above. Libraries were generated by introducing mutations into CDR H1, H2 and H3 and CDR L1 of the antibody GUCY2C-1608 (SEQ ID NO: 221). Phage libraries were rescued as described above and selected using the competitive selection strategy shown in Table 40. For Round 1, phage was incubated with streptavidin particles complexed with biotinylated human GUCY2c (5 nM; SEQ ID NO: 232). After 3 washes, a 100-fold excess of non-biotinylated antigen (500 nM) was added to the reaction mixture and the mixture was rotated overnight at 4°C. The particles were washed the next day to remove phage dissociated from the particles, and the remaining bound phage was eluted using TEA. For round 2, selection was repeated as described above against 0.5 nM antigen without any competition. Colonies were selected and periplasmic preparations were prepared as described above. Clones were screened for improved binding activity using the competitive ELISA described above. Clones that showed improved binding compared to the parental variants were selected for further analysis. Following screening, DNA sequencing was performed as described above on the best clones to identify mutations contributing to improved GUCY2c binding activity.

Following the initial affinity maturation selection library, the CDR regions of these top variants were combined into a new library and additional phage display selections were performed as shown in Table 41. Round 1 selections were performed as described above for the initial affinity maturation selection library, except that that 0.5 nM biotinylated antigen was used. For each library, there were two round 2 selection groups in which the phage was or was not stimulated by incubation at 65°C prior to selection. After phage selection, colonies were selected and periplasmic preparations were prepared as described above. Clones were screened by competitive ELISA described above. Following screening, DNA sequencing of the best clones was performed to identify mutations contributing to improved GUCY2c binding activity. Tables 40 and 41 show the phage display selection strategy used for affinity maturation libraries (C=library; R=round; Dslxn=deselection conditions; Therm=thermal conditions; SA=streptavidin; D=DNA; I=insulin; M=membrane extract).

Table 40

Round Conditions R1 Group C1R1 Target Human GUCY2C Target concentration 5 nM Dslxn SA, D, I, M Therm No Competition 500 nM R2 Group C1R2 Target Human GUCY2C Target concentration 5 nM Dslxn SA, D, I, M Therm No Competition No

Table 41

Round Conditions R1 Group C1R1 Target Human GUCY2C Target concentration 0.5 nM Dslxn SA, D, I, M Therm No Competition 500 nM R2A Group C1R2A Target Human GUCY2C Target concentration 0.5 nM Dslxn SA, D, I, M Therm No Competition 500 nM R2B Group C1R2B Target Man GUCY2C Target concentration 0.5 nM Dslxn SA, D, I, M Therm 65°C Competition 500 nM

The amino acid sequences of anti-GUCY2c antibody clones that demonstrated improved affinity in the PeriPrep competitive ELISA were compared to the parental clone GUCY2C-1608 (VH SEQ ID NO: 73 and VL SEQ ID NO: 147). Changes in amino acid content of the VH CDR that resulted in improved stability are shown in Table 42A. Changes at positions 54, 55, 56, 97, and 99 appear to have the greatest impact on affinity, which correlates with the largest changes in time-resolved fluorescence (according to the Kabat numbering system, using the AbM definition for CDR1 VH).

Table 42A

Designation Position Parent amino acid Optimized Amino Acid CDR1 VH
(H1) H1.31 31 S R, W, Y, A, H, P, Y, T, N, K, D, G, V H1.32 32 Y R, L, T, K, P, I, N, M, V, S CDR2 VH
(H2) H2.52A 52A P T, V H2.53 53 S A, L, R H2.54 54 N T, R, H, K, M, S, A, Y, T, I H2.55 55 E R, K, N, Y, G, L, A, M, S, H, D, Q H2.56 56 L W, Y, F, V, I, N, H H2.57 57 T M, S, L, N, Q, V FW3 VH 93 T A CDR3 VH
(H3) H3.96 96 I F, K H3.97 97 T V, L, I, M, F, Y, A H3.98 98 T N, R, G, L, I H3.99 99 T K, L, W, A, S, M, P, N, R H3.100 100 E G, A, H, S, D, T, R, Q, K, Y, L, M H3.100B 100B Y H H3.100E 100E F L H3.101 101 D Y, E, S

Amino acid content changes in the VL CDR resulting in improved affinity in the PeriPrep competitive ELISA are shown in Table 42B (Chothia numbering system used for VL CDR1). Changes at positions 30, 30a, and 30d in CDR1 VL appear to have the greatest impact on affinity, which correlates with the largest changes in time-resolved fluorescence.

Table 42B

Designation Position Parent amino acid Optimized Amino Acid CDR1 VL
(L1) L1.30 thirty D N, S L1.30a 30a Y W,I L1.30d 30d T S,H

After screening in scFv format as periplasmic preparations, the VH and VL sequences of the best clones were cloned into the full human IgG1/kappa format. The VH and VL genes were amplified by PCR from a phage vector and subcloned into human IgG1 and human kappa vectors by the molecular biology methods described above. The human IgG1 version of these best affinity mature clones was then transiently expressed and purified by the methods also described above. The purified proteins were then analyzed by surface plasmon resonance as described above against recombinant human GUCY2c protein (SEQ ID NO: 232). Table 43 shows the forward reaction rate (ka), reverse reaction rate (kd) and equilibrium constant ( _KD ) for affinity mature clones compared to parental GUCY2C-1608 in IgG form (GUCY2c-1640; SEQ ID NOs: 214 and 223 ). Several variants exhibited 3.4-fold improved affinity compared to the parent antibody. Table 44 shows the complete VH and VL amino acid sequences of these affinity mature versions of GUCY2C-1608.

Table 43. Surface plasmon resonance of affinity mature anti-GUCY2c IgG relative to the human GUCY2c extracellular domain (ECD) protein.

Ligand ka (1/Ms) kd (1/s) K _D (nM) GUCY2C-1640 1.95E+05 2.71E-03 13.87±0.38 GUCY2C-1696 1.44E+05 1.79E-03 12.42±0.55 GUCY2C-1701 2.35E+05 2.79E-03 11.85±0.38 GUCY2C-1702 2.46E+05 1.51E-03 6.16±0.34 GUCY2C-1703 2.10E+05 2.14E-03 10.32±1.34 GUCY2C-1705 2.39E+05 1.49E-03 6.25±0.04 GUCY2C-1706 1.98E+05 1.93E-03 9.75±0.07 GUCY2C-1708 1.40E+05 1.32E-03 9.44±0.4 GUCY2C-1710 2.48E+05 1.47E-03 5.94±0.24 GUCY2C-1713 1.61E+05 1.08E-03 6.71±0.33 GUCY2C-1714 1.21E+05 1.24E-03 10.27±0.59 GUCY2C-1715 2.72E+05 1.10E-03 4.07±0.4

Table 44. Amino acid sequences of the affinity mature regions VH and VL

SEQ ID NO: 294 GUCY2C-1696_VH EVQLVESGGGLVQPGGSLRLSCAASGTFSTRWMHWVRQAPGKGLEWIGEIKPSNKLTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARTITTTEGYWFLSDWGQGTLVTVSS SEQ ID NO: 295 GUCY2C-1696_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGHSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 296 GUCY2C-1701_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTPLGYWFFDVWGQGTLVTVSS SEQ ID NO: 297 GUCY2C-1701_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 298 GUCY2C-1702_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTPLGYWFFDVWGQGTLVTVSS SEQ ID NO: 299 GUCY2C-1702_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 300 GUCY2C-1703_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSNRWNNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS SEQ ID NO: 301 GUCY2C-1703_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 302 GUCY2C-1705_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSRGFTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS SEQ ID NO: 303 GUCY2C-1705_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 304 GUCY2C-1706_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSTWMHWVRQAPGKGLEWIGEIKPSNELTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTEGYWFFDVWGQGTLVTVSS SEQ ID NO: 305 GUCY2C-1706_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 306 GUCY2C-1708_VH EVQLVESGGGLVQPGGSLRLSCAASGTFFSRPWMHWVRQAPGKGLEWIGEIKPSTGWTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTTGYWFFDVWGQGTLVTVSS SEQ ID NO: 307 GUCY2C-1708_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYKHSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 308 GUCY2C-1710_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSRGWTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARTITTTEGYWFFDVWGQGTLVTVSS SEQ ID NO: 309 GUCY2C-1710_VL DIQLTQSPSSSLSASVGDRVTITCRASESVDYYRHSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 310 GUCY2C-1713_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSHTWMHWVRQAPGKGLEWIGEIKPSRGFTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTITTTQGYWFFDVWGQGTLVTVSS SEQ ID NO: 311 GUCY2C-1713_VL DIQLTQSPSSSLSASVGDRVTITCRASESVNWYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 312 GUCY2C-1714_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSHTWMHWVRQAPGKGLEWIGEIKPSTKYTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCARTIFTREGYWFFDVWGQGTLVTVSS SEQ ID NO: 313 GUCY2C-1714_VL DIQLTQSPSSSLSASVGDRVTITCRASESVNWYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK SEQ ID NO: 314 GUCY2C-1715_VH EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMHWVRQAPGKGLEWIGEIKPSTKYTNVHEKFKDRFTISVDKAKNSAYLQMNSLRAEDTAVYYCTRTIFTNEGYWFFDVWGQGTLVTVSS SEQ ID NO: 315 GUCY2C-1715_VL DIQLTQSPSSSLSASVGDRVTITCRASESVSIYGSSLMQWYQQKPGKAPKLLIYAASKLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQTRKAYTFGQGTKLEIK

Example 29: Determination of Epitopes by Peptide Mapping of the GUCY2C Extracellular Domain

Peptide mapping was performed to determine the epitope on the extracellular domain (ECD) of GUCY2c to which the primary anti-GUCY2c antibody binds. This approach produced 20-mer peptides with a 15 amino acid overlap spanning the entire extracellular domain in the GUCY2c transmembrane domain consisting of amino acids 1-440 (Table 45). Peptides were synthesized and provided as panels in lyophilized format (New England Peptide, Gardner MA). The panels were re-diluted to 100 μg/ml with acetonitrile and then further diluted to 20 μg/ml with sodium carbonate/bicarbonate buffer. Purified antibodies or bispecific antibodies against GUCY2c-CD3 were then tested by Delfia ELISA for binding to various peptides using the Delfia ELISA methods described above, with diluted peptides used as target antigens. GUCY2C-1608 exhibited strong binding to peptides 19 and 20 (SEQ ID NOs: 334 and 335), as measured by time-resolved fluorescence (TRF). Binding data from the TRF results for GUCY2C-1608 relative to all peptides are shown in Table 45. The sequence NSGDCRSSTCEGLDLLRKISNAQRM (SEQ ID NO: 405) is covered by these peptides. The peptide overlap region is RSSTCEGLDLLRKIS (SEQ ID NO: 406), indicating an epitope in the region.

Table 45

Seq ID Peptide no. Peptide sequence TRF DELFIA 316 1 MKTLLLDLALWSLLFQPGWL 166.25 317 2 LDLALWSLLFQPGWLSFSSQ 106.5 318 3 WSLLFQPGWLSFSSQVSQNC 97.75 319 4 QPGWLSFSSQVSQNCHNGSY 124.75 320 5 SFSSQVSQNCHNGSYEISVL 108.75 321 6 VSQNCHNGSYEISVLMMGNS 412.5 322 7 HNGSYEISVLMMGNSAFAEP 136 323 8 EISVLMMGNSAFAEPLKNLE 147.5 324 9 MMGNSAFAEPLKNLEDAVNE 145 325 10 AFAEPLKNLEDAVNEGLEIV 105.25 326 eleven LKNLEDAVNEGLEIVRGRLQ 101.25 327 12 DAVNEGLEIVRGRLQNAGLN 258.75 328 13 GLEIVRGRLQNAGLNVTVNA 101.5 329 14 RGRLQNAGLNVTVNATFMYS 97 330 15 NAGLNVTVNATFMYSDGLIH 88.75 331 16 VTVNATFMYSDGLIHNSGDC 105.25 332 17 TFMYSDGLIHNSGDCRSSTC 112 333 18 DGLIHNSGDCRSSTCEGLDL 126 334 19 NSGDC RSSTCEGLDLLRKIS 19179.25 335 20 RSSTCEGLDLLRKIS NAQRM 3208.75 336 21 EGLDLLRKISNAQRMGCVLI 81 337 22 LRKISNAQRMGCVLIGPSCT 93.5 338 23 NAQRMGCVLIGPSCTYSTFQ 96.25 339 24 GCVLIGPSCTYSTFQMYLDT 118.75 340 25 GPSCTYSTFQMYLDTELSYP 131 341 26 YSTFQMYLDTELSYPMISAG 104.25 342 27 MYLDTELSYPMISAGSFGLS 105 343 28 ELSYPMISAGSFGLSCDYKE 96.75 344 29 MISAGSFGLSCDYKETLTRL 101 345 thirty SFGLSCDYKETLTRLMSPAR 125 346 31 CDYKETLTRLMSPARKLMYF 90.25 347 32 TLTRLMSPARKLMYFLVNFW 83.25 348 33 MSPARKLMYFLVNFWKTNDL 87.5 349 34 KLMYFLVNFWKTNDLPFKTY 99 350 35 LVNFWKTNDLPFKTYSWSTS 108 351 36 KTNDLPFKTYSWSTSYVYKN 111.5 352 37 PFKTYSWSTSYVYKNGTETE 106.75 353 38 SWSTSYVYKNGTETEDCFWY 109.5 354 39 YVYKNGTETEDCFWYLNALE 86.5 355 40 GTETEDCFWYLNALEASVSY 85.25 356 41 DCFWYLNALEASVSYFSHEL 161 357 42 LNALEASVSYFSHELGFKVV 96.5 358 43 ASVSYFSHELGFKVVLRQDK 104 359 44 FSHELGFKVVLRQDKEFQDI 90.5 360 45 GFKVVLRQDKEFQDILMDHN 107.5 361 46 LRQDKEFQDILMDHNRKSNV 105.75 362 47 EFQDILMDHNRKSNVIMCG 103.75 363 48 LMDHNRKSNVIIMCGGPEFL 148 364 49 RKSNVIIMCGGPEFLYKLKG 129.25 365 50 IIMCGGPEFLYKLKGDRAVA 161 366 51 GPEFLYKLKGDRAVAEDIVI 96.75 367 52 YKLKGDRAVAEDIVIILVDL 103.75 368 53 DRAVAEDIVIILVDLFNDQY 87.75 369 54 EDIVIIILVDLFNDQYLEDNV 112.25 370 55 ILVDLFNDQYLEDNVTAPDY 100.25 371 56 FNDQYLEDNVTAPDYMKNVL 81 372 57 LEDNVTAPDYMKNVLVLTLS 99.75 373 58 TAPDYMKNVLVLTLSPGNSL 98.5 374 59 MKNVLVLTLSPGNSLLNSSF 135.25 375 60 MKNVLVLTLSPGNSLLNSSF 150.25 376 61 VLTLSPGNSLLNSSFSRNLS 125.5 377 62 PGNSLLNSSFSRNLPTKRD 91.25 378 63 LNSSFSRNLSPKRDFALAY 109.5 379 64 SRNLSPTKRDFALAYLNGIL 116 380 65 PTKRDFALAYLNGILLFGHM 103 381 66 FALAYLNGILLFGHMLKIFL 114.25 382 67 LNGILLFGHMLKIFLENGEN 93.25 383 68 LFGHMLKIFLENGENITTPK 73.5 384 69 LKIFLENGENITTPKFAHAF 100.5 385 70 ENGENITTPKFAHAFRNLTF 91.75 386 71 ITTPKFAHAFRNLTFEGYDG 105 387 72 FAHAFRNLTFEGYDGPVTLD 116 388 73 RNLTFEGYDGPVTLDDWGDV 116.25 389 74 EGYDGPVTLDDWGDVDSTMV 100 390 75 PVTLDDWGDVDSTMVLLYTS 88.25 391 76 DWGDVDSTMVLLYTSVDTKK 102 392 77 DSTMVLLYTSVDTKKYKVLL 98.25 393 78 LLYTSVDTKKYKVLLTYDTH 97.75 394 79 VDTKKYKVLLTYDTHVNKTY 130 395 80 YKVLLTYDTHVNKTYPVDMS 1468.75 396 81 TYDTHVNKTYPVDMSPTFTW 109 397 82 VNKTYPVDMSPTFTWKNSKL 99.25 398 83 PVDMSPTFTWKNSKLPNDIT 85.75 399 84 PTFTWKNSKLPNDITGRGPQ 133 400 85 KNSKLPNDITGRGPQILMIA 163.25 401 86 PNDITGRGPQILMIAVFTLT 110.5 402 87 LRKISNAQRMGCVLIGPSCT 102 403 88 NAQRMGCVLIGPSCTYSTFQ 120.25 404 89 GCVLIGPSCTYSTFQMYLDT 115 405 NSGDCRSSTCEGLDLLRKISNAQRM 406 RSSTCEGLDLLRKIS

GUCY2c peptide 19 (SEQ ID NO: 334) was then determined to compete with the entire extracellular domain in a DELFIA competitive ELISA using the methods described above and was shown to compete effectively with the entire extracellular domain (FIG. 19). This suggests that peptide 19 (SEQ ID NO 334) contains the specific epitope that GUCY2C-1608 binds to on the entire extracellular domain of GUCY2c.

GUCY2C-1608 binds to a different epitope of GUCY2c ECD than Antibody MS20

The binding of GUCY2C-1608 to the extracellular domain of GUCY2c was also compared to that known with anti-GUCY2c antibodies. The MS20 antibody described in US2013/315923 does not have a described epitope. To compare this antibody with GUCY2C-1608, a Delfia competitive ELISA was performed using a modified Delfia competitive ELISA method described above. In this case, human GUCY2c protein (SEQ ID NO: 232) was coated onto the plate as described above. GUCY2C-1640 (IgG version of GUCY2C-1608; SEQ ID NO: 214 and 223) or MS20 antibody was serially diluted starting at 20 μg/ml, then combined with the chimeric rabbit IgG version of GUCY2C-0098 (SEQ ID NO: 440 and 441, consisting of SEQ ID NOs: 26 and 106 fused to a rabbit constant domain) in an amount of 250 ng/ml (EC ₅₀ value determined separately). The plates were washed as described above, and a secondary anti-rabbit europium antibody (Perkin Elmer) diluted 1:1000 was added for detection. TRF signal was determined as previously described. A decrease in the TRF signal indicates competitive binding, because the chimeric rabbit version GUCY2C-0098 is displaced from binding to human GUCY2c. As expected, GUCY2C-1640 displaced the chimeric rabbit version GUCY2C-0098, while MS20 did not (Figure 20). Thus, GUCY2C-1608 binds to a different epitope of the GUCY2c ECD than the MS20 antibody.

Example 30: GUCY2c-1608 binds to a different GUCY2c ECD epitope than antibody 5F9

The antibodies described in US2011/0110936 are described to bind to different epitope sequences on the GUCY2c ECD from peptide 19 (SEQ ID NO 334), as presented herein below.

3×10 ⁶ cells expressing human, rat and chicken GUCY2c (CID1814, 1815 and 1818, respectively), point mutants CID1941-1950, patch mutants CID1868-1873 (patch mutants consist of 2-5 amino acid residues of human GUCY2c replaced by amino acid residues from equivalent positions in chicken GUCY2c), or human-chicken GUCY2c chimeras CID1874-1881 (>5 amino acids of human GUCY2c replaced by chicken GUCY2c residues), or confirmatory reverse chimeras CID1939 and 1940 (> 5 chicken GUCY2c amino acids replaced with human GUCY2c residues), or the negative control CID1431 (a negative control construct that does not contain the GUCY2s sequence but has the same epitope tags as other constructs expressed on the surface of yeast). Cells were incubated with anti-V5 antibody (Abcam 27671), monovalent or bivalent anti-GUCY2c antibodies, GUCY2C-1608 antibody, or 5F9 antibody (Millennium antibody described in US2011/0110936) for 1 hour at room temperature. Cells were then washed three times with PBS+0.5% BSA buffer. Cells were then incubated with PE-conjugated anti-mouse IgG or PE-conjugated anti-human IgG or PE-conjugated anti-HA (Miltenyi Biotec 130-092-257) for 30 minutes at 4°C in the dark. Cells were washed three times with PBS+0.5% BSA buffer and then analyzed using a flow cytometer.

The MFI (median fluorescence intensity) of anti-GUCY2c antibody binding to cells was normalized to cell surface expression measured by the MFI of anti-tag antibodies. All cells expressed V5 and HA on the cell surface to normalize expression. Table 46 shows the normalized relative binding of anti-GUCY2c antibodies to each GUCY2c construct. Chimeras (except CID1939 and CID1940), point mutants, and patch mutants that do not bind to the anti-GUCY2c Ab demonstrate that the GUCY2c residues altered in these constructs are important for antibody binding. Restoration of binding of GUCY2C-1608, but not antibody 5F9, to the reverse chimeras CID1939 and CID1940 confirms that antibody GUCY2C-1608 binds to residues 68-87 on GUCY2c (SEQ ID NO: 224) and suggests that the binding site of antibody 5F9 is not overlaps with the binding site of the antibody GUCY2C-1608.

Table 46

Construction of GUCY2c (SEQ ID) Ab GUCY2Cc-1608 Ab 5F9 CID1814 (410) + + CID1815 (411) + - CID1818 (412) - - CID1868 (413) - + CID1869 (414) + + CID1870 (415) + + CID1871 (416) + + CID1872 (417) + + CID1873 (418) + + CID1874 (419) - + CID1875 (420) + + CID1876 (421) + + CID1877 (422) + + CID1878 (423) + + CID1879 (424) + + CID1880 (425) + + CID1881 (426) + + CID1939 (427) + - CID1940 (428) + - CID1941 (429) + + CID1942 (430) + + CID1943 (431) + + CID1944 (432) + + CID1945 (433) - + CID1946 (434) + + CID1947 (435) + + CID1948 (436) + + CID1949 (437) + + CID1950 (438) + + CID1431 (439) - -

Example 31: Crystallization and structure determination of scFv complex against GUCY2C-1608 and GUCY2c peptide

For crystallization studies, a complex of scFv GUCY2C-1608 (SEQ ID NO 409) and ECD peptide GUCY2c No. 19 ⁶⁸ NSGDCRSSTCEGLDLLRKIS ⁸⁷ (SEQ ID NO 334) was prepared in a molar ratio of 1:1.2 and concentrated to 8.8 mg/ml in a protein solution , containing TBS, at pH 7.5. Crystals were prepared by hanging drop vapor diffusion under conditions containing 20% PEG 3350, 200 mM lithium sulfate, bis-Tris, pH 5.5. The rod crystals had symmetry corresponding to the monoclinic space group = P2 ₁ 2 ₁ 2 ₁ with cell parameters a = 70.68 Å; b=80.57 Å; c=90.7 Å and with two copies of scFv GUCY2C-1608 peptide complexes in a crystallographic asymmetric unit. The crystals were cryopreserved using a reservoir solution containing 20% ethylene glycol and quickly frozen in liquid nitrogen. A data set to 1.6 Å resolution was obtained from a single frozen crystal using the IMCA 17-ID synchrotron radiation beam at Argonne National Laboratory (APS). Data were processed and scaled using autoPROC, and the final dataset was 71.4% complete.

The structure was resolved by molecular replacement using PHASER, starting with a single-chain Fv fragment derived from the structure of a bispecific antibody against GUCY2c and CD3 (Figure 22). Resolution was achieved by searching for two copies of scFv GUCY2C-1608. The resulting electron density maps, calculated using two copies of scFv GUCY2C-1608 as a model, clearly show additional electron densities for two peptides, each bound to a corresponding copy of scFv GUCY2C-1608.

Through several iterative rounds of manual adjustments and model rebuilding using COOT and crystallographic refinement using autoBUSTER, a final GUCY2C-1608 scFv+peptide model with crystallographic R _{working values was obtained.} 19.7% and R _free. 21.7%, where R _working. = ||F _obs. |-|F _calc. ||/|F _obs. |, and R _free is equivalent to R _working. , but calculated for a randomly selected 5% of reflections omitted during refinement.

A. Description of the structure of scFv GUCY2C-1608-peptide

The 17-residue GUCY2c peptide adopts a primarily α-helical conformation upon binding to the CDR regions of the GUCY2C-1608 scFv (Figure 21). The α-helix is held by an SS disulfide bond to the N-terminus of the peptide. The total binding surface area submerged relative to the interaction region is ~1.320 Å ² , which is unexpectedly large for a fragment only 17 amino acid residues long. The binding surface is mainly hydrophobic with some specific polar contacts in the center and hydrogen bond contacts at the periphery (Figure 21).

Binding contacts of epitopes within 3.8 Å of scFv GUCY2C-1608 are identified by underlined residues in the ⁶⁸ NS GDCRSSTCEGLDLLRKI S ⁸⁷ peptide sequence and are listed in Table 47. Amino acids comprising >75% of the binding surface area are indicated in bold. The binding paratope scFv GUCY2C-1608 is defined by 5 CDR regions: CDR-H1, CDR-H2, CDR-H3, CDR-L1 and CDR-L3. The amino acids of scFv GUCY2C-1608 (paratope) and GUCY2c-peptide (epitope) contributing to contact within 3.8 Å are given in Table 48.

Table 47. GUCY2c epitope residues

Type Chain GUCY2c Position %BSA Electrostatic interactions A.R.G. C 73 50.28431 H SER C 74 56.99118 SER C 75 93.22802 H THR C 76 85.61988 H G.L.U. C 78 46.89699 H GLY C 79 86.96493 W LEU C 80 78.57378 LEU C 82 28.2139 LEU C 83 88.59774 A.R.G. C 84 40.15275 ILE C 86 23.20466

Table 48. Amino acids that determine the paratope (H+L chains) and epitope (C chain)

Residues having atoms within 3.80 Å Antigen chain Antigen residue Antibody chain Antibody residue (consistent) (Kabat) C 73 H 33 33 C 73 H 57 56 C 73 H 58 57 C 73 H 59 58 C 74 H 33 33 C 74 H 50 50 C 74 H 59 58 C 75 H 33 33 C 75 H 50 50 C 75 L 99 96 C 76 L 96 92 C 76 L 98 94 C 78 H 33 33 C 79 H 107 100C C 80 H 107 100C C 80 L 31 27D C 80 L 96 92 C 82 H 101 97 C 83 H 104 100 C 83 H 105 100A C 83 L 36 32 C 84 L 31 27D C 84 L 32 28 C 86 H 103 99 C 86 H 104 100 C 86 H 105 100A

B. Crystallization and structure determination of bispecific antibodies against GUCY2c and CD3

For crystallization studies, a modified version of GUCY2C-1608 was prepared with a His6 tag or a FLAG tag at the C terminus of each VL-VH subunit strand to facilitate purification. Bispecific antibody against GUCY2c and CD3 for crystallography was prepared using the sequences VH GUCY2C-1637 (SEQ ID NO: 407) and VL GUCY2C-1637 (SEQ ID NO: 408). The constructs were transiently transfected into FreeStyle™ 293 HEK cells as described above. After collecting the conditioned media, 2.5 mL of Ni-NTA resin, pre-equilibrated with TBS, was allowed to bind to protein for 1 hour at 4°C using an orbital mixer. The resin was then collected and placed on an Applied Biosystems column (Life Technologies, Grand Island, NY, USA). The resin was first washed to its original level with buffer A (50 mM sodium phosphate, 300 mM sodium chloride, pH 8.0), then with five CVs of buffer A supplemented with 20 mM imidazole, and finally eluted with buffer A+250 mM imidazole. Fractions with the highest purity were pooled and dialyzed into PBS using a 30 ml cassette with a molecular weight cut-off of 10 kDa for 2 hours at 4°C. The dialyzed protein was then further purified by chromatography using a FLAG tag. Forty mL of anti-FLAG tag M2 resin (Sigma, St. Louis, MO, USA) was pre-equilibrated with TBS and allowed to bind in 1.4 L of conditioned medium overnight at 4°C. The resin was then collected and packed onto an M2 anti-FLAG chromatography column, washed to stock using TBS (20 CV) and first eluted with 0.1 M FLAG peptide buffer and finally eluted with 0.1 M glycine pH 3.0. The eluted protein was immediately neutralized with 10% 1.0 M Tris, pH 8.0. Fractions with the highest purity were then pooled and further purified by size exclusion chromatography using a Superdex200 column (GE Healthcare, Piscataway, NJ, USA) and stored in TBS.

For crystallization studies, the His/FLAG-purified version of GUCY2C-1608 was concentrated to 12 mg/mL in a protein solution containing TBS, pH 7.5. Crystals were prepared by the hanging drop method by vapor diffusion under conditions including 1 M sodium malonate, pH 6, 2% ethyleneimine polymer. The crystals had symmetry corresponding to the hexagonal space group=P3 ₂ 2 ₁ with cell parameters a=b=119.55 Å and c=121.85 Å and with one copy of the diabody in the crystallographic asymmetric unit. The crystals were cryopreserved using a reservoir solution containing 2.8 M sodium malonate, pH 6, and flash frozen in liquid nitrogen. A data set to 2.03 Å resolution was obtained from a single frozen crystal using the IMCA 17-ID synchrotron radiation beam at Argonne National Laboratory (APS). Data were processed and scaled using autoPROC, and the final dataset was 49.3% complete.

The structure was resolved by molecular replacement using PHASER, starting with single-chain Fv fragment models obtained by Brookhaven PDB entry 1moe. Resolution was achieved through a separate search for each of the four subunits of the diabody molecule. Through several iterative rounds of manual adjustments and model rebuilding using COOT and crystallographic refinement using autoBUSTER, a final diabody model with crystallographic R _{working values was obtained.} 19.1% and R _free. 20.8%, where R _working. = ||F _obs. |-|F _calc. ||/|F _obs. |, and R _{is free.} equivalent to R _working. , but calculated for a randomly selected 5% of reflections omitted during refinement.

C. Description of the structure of the anti-GUCY2C-CD3 diabody

The two antigen binding sites on the bispecific antibody against GUCY2c and CD3 are separated by approximately 70 Å and are on opposite sides of the molecule, directly opposite each other (Figure 22). The anti-CD3 CDR and anti-GUCY2c CDR regions are located at a distance from the subunit contact region.

Example 32: Evaluation in vivo combinations of bispecific antibody against GUCY2c-CD3 with mAb against VEGF-A and axitinib

Human PBMCs were thawed in media (X-VIVO 15 (Lonza). 5% human serum albumin (Gemini #100-318), 1% penicillin/streptomycin, 0.01 mM 2-mercaptoethanol was added to approximately 5 million cells per ml. Cells centrifuged and resuspended in Robosep buffer (Stem Cell Technologies) at a concentration of 50 million cells per ml T cells were isolated using the EasySep human T cell enrichment kit (Stem Cell Technologies) T cells were activated and grown using the activation kit /human T cell expansion (Miltenyi) On day 2, T cells were transferred to a G-Rex cell culture apparatus for expansion and the medium was supplemented with IL-2 (Stem Cell Technologies) and replenished 2 days later. 1 week after activation/expansion At the time of collection, particles were removed using a magnet and cells were resuspended in DPBS at 1 x 10 ⁷ cells/ml for in vivo inoculation.

NSG mice were inoculated subcutaneously in the flank with patient-derived colorectal xenograft fragments PDX-CRX-11201. Tumors were measured using a digital caliper and volumes were calculated using a modified ellipsoid formula of ½ × length × width2. Mice were randomized and staged according to tumor size of 150-200 mm ³ . An initial dose of anti-GUCY2c bispecific antibody (GUCY2C-1608), negative control bispecific antibody, or vehicle was administered to animals on day 0, and 2 million cultured T cells were inoculated the next day. Mice were dosed as bolus injections of 0.2 ml weekly up to 3 times, and all compound and T cell administrations were intravenous through the tail vein of each animal. The combination agents were administered starting on day 0. Anti-VEGF mAb (G6-31) was administered every 3 days up to 4 times intravenously via the tail vein. Axitinib was administered twice daily for up to 14 days by gastric tube. Tumors were measured twice weekly along with continuous monitoring for signs of graft-versus-host disease.

A bispecific antibody against GUCY2c and CD3 administered at 0.05 mg/kg demonstrated a partial antitumor response in the PDX-11201 adoptive transfer model. Anti-VEGF mAb and axitinib also demonstrated partial antitumor responses in this model when administered alone at 5 mg/kg and 15 mg/kg, respectively. Combination of bispecific antibody against GUCY2c and CD3 with anti-VEGF mAb or axitinib resulted in additive antitumor responses demonstrating complete tumor inhibition.

GUCY2C-1608 demonstrated enhanced tumor growth inhibition when combined with an anti-VEGF mAb as well as axitinib in the patient-derived colorectal carcinoma xenograft model PDX-CRX-11201 with adoptive T-cell transfer.

Example 33: Evaluation in vivo combinations of bispecific antibodies against GUCY2c and CD3 with an antibody against PD1

Human PBMCs were thawed in media (X-VIVO 15 (Lonza). 5% human serum albumin (Gemini #100-318), 1% penicillin/streptomycin, 0.01 mM 2-mercaptoethanol was added to approximately 5 million cells per ml. Cells centrifuged and resuspended in Robosep buffer (Stem Cell Technologies) at a concentration of 50 million cells per ml T cells were isolated using the EasySep human T cell enrichment kit (Stem Cell Technologies) T cells were activated and grown using the activation kit /human T cell expansion (Miltenyi) On day 2, T cells were transferred to a G-Rex cell culture apparatus for expansion, media were supplemented with IL-2 (Stem Cell Technologies) and replenished 2 days later. 1 week after activation/expansion At the time of collection, particles were removed using a magnet and cells were resuspended in DPBS at 1 x 10 ⁷ cells/ml for in vivo inoculation.

NSG mice were inoculated subcutaneously in the flank with colorectal cancer cell line LS1034. Tumors were measured using a digital caliper and volumes were calculated using a modified ellipsoid formula of ½ × length × width2. Mice were randomized and staged according to tumor size of 150-200 mm ³ . An initial dose of bispecific antibody against GUCY2c (GUCY2C-1608) in the amount of 0.03 mg/kg, bispecific antibody of negative control in the amount of 0.1 mg/kg, antibody against PD1 (biosimilar to pembrolizumab D265A, effector function mutant Fc hIgG1a) in the amount 5 mg/kg or vehicle was administered to animals on day 0 and 2 million cultured T cells were inoculated the next day. Mice were dosed as bolus injections of 0.2 ml weekly up to 3 times, and all compound and T cell administrations were intravenous through the lateral tail vein of each animal. Tumors were measured twice weekly along with continuous monitoring for signs of graft-versus-host disease.

A bispecific antibody against GUCY2c and CD3 administered at 0.03 mg/kg demonstrated a partial antitumor response in the LS1034 adoptive transfer model. Anti-PD-1 antibody did not demonstrate an antitumor response in this model when administered at 10 mg/kg. Combining a bispecific antibody against GUCY2c and CD3 with an antibody against PD-1 resulted in an additive antitumor response (Figure 24).

GUCY2C-1608 demonstrated enhanced tumor growth inhibition when combined with anti-PD1 mAb in the colorectal carcinoma cell line-derived xenograft LS1034 T-cell adoptive transfer model.

Example 34: Evaluation in vivo combinations of bispecific antibodies against GUCY2c and CD3 with an antibody against PDL1

Human PBMCs were thawed in media (X-VIVO 15 (Lonza). 5% human serum albumin (Gemini #100-318), 1% penicillin/streptomycin, 0.01 mM 2-mercaptoethanol was added to approximately 5 million cells per ml. Cells centrifuged and resuspended in Robosep buffer (Stem Cell Technologies) at a concentration of 50 million cells per ml T cells were isolated using the EasySep human T cell enrichment kit (Stem Cell Technologies) T cells were activated and grown using the activation kit /human T cell expansion (Miltenyi) On day 2, T cells were transferred to a G-Rex cell culture apparatus for expansion, media were supplemented with IL-2 (Stem Cell Technologies) and replenished 2 days later. 1 week after activation/expansion At the time of collection, particles were removed using a magnet and cells were resuspended in DPBS at 1 x 10 ⁷ cells/ml for in vivo inoculation.

NSG mice were inoculated subcutaneously in the flank with colorectal cancer cell line LS1034. Tumors were measured using a digital caliper and volumes were calculated using a modified ellipsoid formula of ½ × length × width2. Mice were randomized and staged according to tumor size of 150-200 mm ³ . An initial dose of 0.03 mg/kg anti-GUCY2c bispecific antibody (GUCY2C-1608) or 0.1 mg/kg negative control bispecific antibody or vehicle was administered to the animals on day 0 and 2 million cultured T cells were inoculated onto the next day. Mice were dosed as bolus injections of 0.2 ml weekly up to 3 times, and all compound and T cell administrations were intravenous through the lateral tail vein of each animal. Anti-PDL1 antibody was administered intravenously starting on day 0, every 3 days up to 6 times. Tumors were measured twice weekly along with continuous monitoring for signs of graft-versus-host disease.

A bispecific antibody against GUCY2c and CD3 administered at 0.03 mg/kg demonstrated a partial antitumor response in the LS1034 adoptive transfer model. Anti-PD-L1 antibody did not demonstrate an antitumor response in this model when administered at 10 mg/kg. Combining a bispecific antibody against GUCY2c and CD3 with an antibody against PD-L1 resulted in an additive antitumor response (Figure 25).

GUCY2C-1608 demonstrated enhanced tumor growth inhibition when combined with anti-PD-L1 mAb in the colorectal carcinoma cell line-derived xenograft LS1034 T-cell adoptive transfer model.

Example 35: Phase 1 Dose Escalation and Expansion Population Study of a Previously Established Maximum Tolerated Dose to Evaluate the Safety, Tolerability, Pharmacokinetics, Pharmacodynamics, and Antitumor Activity of the GUCY2c-1608 Bispecific Antibody in Patients with Advanced Gastrointestinal Tumors

This example describes an open-label, multi-center, multi-dose Phase 1 safety, PK and PD study of the bispecific antibody GUCY2c-1608. The study consists of two parts, dose escalation as a single agent and dose escalation in combination (with anti-PD-1 antibody or anti-VEGF antibody) (Parts 1A and 1B, respectively) followed by a study of the previously established maximum tolerated dose in an expansion population (Part 2 ). Consecutive cohorts of patients with advanced/metastatic gastrointestinal tumors, including colorectal adenocarcinoma, gastric and esophageal adenocarcinoma, who were not candidates for treatment regimens known to provide clinical benefit, will be administered escalating doses of the bispecific antibody GUCY2c-1608 at Part 1A of the study. Part 1B will dose titrate the bispecific antibody GUCY2c-1608 in combination (with anti-PD-1 or anti-VEGF) in patients with colorectal adenocarcinoma, advanced/metastatic adenocarcinomas of the stomach and esophagus. Part 2 (phase study of the previously established maximum tolerated dose in the expansion population) will evaluate the recommended phase 2 dose (RP2D) selected in Part 1 (dose escalation phase), either as monotherapy or in combination (with anti-PD-1 antibody or anti-VEGF) in patients with previously treated colorectal adenocarcinomas. Cohorts to evaluate biomarkers that require paired tumor biopsies from these patients can be used in parts 1A and 1B as the maximum tolerated dose (MTD) approaches or in the case of RP2D/MTD and in subgroups of the previously established maximum tolerated dose study arms in an expanded population in parts 2.

Increasing doses of GUCY2c-1608 will be administered as weekly subcutaneous (SC) injections in 21-day cycles. The suggested starting dose for SC is 0.6 mcg/kg.

Once GUCY2c-1608 RP2D/MTD is identified as monotherapy, Part 1B dose cohorts will be assembled to study safety, tolerability, and preliminary antitumor activity at this dose level in combination (with anti-PD-1 antibody or anti-VEGF antibody). The previously established maximum tolerated dose (MTD) expansion population phase (Part 2) will evaluate RP2D GUCY2c-1608 as monotherapy and combination therapy (with anti-PD-1 antibody or anti-VEGF antibody) in patients with advanced/metastatic colorectal adenocarcinomas.

There will also be at least a 72-hour interval between the first dose administered to each of the original patients (ie, patients included in the original dose-limiting toxicity (DLT) assessment) enrolled in the study at the new dose level. All patients will be observed as an inpatient for at least 48 hours or more after the first SC dose of Cycle 1 Day 1 (C1D1) Part 1.

Treatment with GUCY2c-1608 will continue until disease progression, patient failure, or unacceptable toxicity, whichever occurs first, unless the investigator and medical monitor agree to treat in the setting of disease progression based on an individual benefit/risk assessment.

Alternative dosing regimen with IV doses

Intravenous (IV) administration can be evaluated based on available clinical data (including safety/tolerability, PK, PD, etc.) obtained from dose escalation cohorts with SC administration.

This study may evaluate an intensified initial dose to allow subsequent higher dose levels.

Criteria for dose escalation

For administration cohorts, initial dose levels are shown in Table 49; intermediate doses can be assessed based on clinical results. For SC administration cohorts, during initial dose escalation levels, maximum dose escalations will be up to 250%, but will be adjusted to no more than 100% after DLT observation.

Table 49

SC dose escalation levels for administration Dose level Dose (mcg/kg) DL1 (Initial dose) 0.6 DL2 2.0 DL3 and higher Continued enhancement to MTD or desired pharmacological activity

*Intermediate doses may be assessed based on clinical results

Dose increases will be stopped when stopping criteria are reached, as determined by one skilled in the art.

Part 1A of the study is designed to determine the MTD for the SC dosing regimen, however, if the dosing regimen is switched to IV, the MTD for IV dosing will be determined. If inclusion of a boosted starting dose is required for an IV, the MTD for the IV will be determined using the boosted starting dose schedule.

Part 1B will be designed to determine the MTD of GUCY2c-1608 as a combination agent (with anti-PD-1 antibody or anti-VEGF antibody).

RP2D is the dose selected for further study based on the results of the phase 1 study.

Patient selection

For dose escalation (Parts 1A and 1B): histologic or cytologic diagnosis of colorectal adenocarcinoma or advanced/metastatic gastric or esophageal adenocarcinoma that is refractory to standard therapy or for which standard therapy is not available.

For a previously established maximum tolerated dose study in an expansion population (Part 2): histologic or cytologic diagnosis of previously treated colorectal adenocarcinoma with at least one measurable lesion not previously irradiated, as defined by RECIST version 1.1, that is refractory to standard therapy , or for which standard therapy is not available.

Treatment of cytokine release syndrome

Treatment of cytokine release syndrome (CRS) and the revised CRS scoring system are adapted from DWLee, et al: Current Concepts in the Diagnosis and Management of Cytokine Release Syndrome. Blood 124 (2014) 188-195.

Anti-IL-6 antibody administration: Consider 4 mg/kg over 1 hour and a second dose if there is no clinical improvement within 24 to 48 hours.

The description of each patent, patent application and publication cited herein is hereby incorporated by reference in its entirety.

Equivalent

Although the present invention has been described with reference to specific embodiments, it will be appreciated that those skilled in the art may develop other embodiments and variations of the present invention without departing from the spirit and scope of the invention. The subject matter of the invention includes all of these other embodiments and variations, including combinations and subcombinations of the various elements, features, functions and/or properties presented herein. The following claims specifically identify certain combinations and sub-combinations that are considered novel and non-obvious. The invention embodied in other combinations and subcombinations of features, functions, elements and/or properties may be claimed in this application, in applications claiming priority under this application, or in related applications. Such claims directed to another invention or the same invention and having a broader, narrower, equal or different scope than the original claims are also considered to be included within the scope of the present invention.

--->

LIST OF SEQUENCES

<110> PFIZER INC.

Chang, Chew

Guntas, Gurkan

Katragadda, Madan

Mathur, Divya

Root, Adam

Mosyak, Lidia

LaVallie, Edward

<120> ANTIBODIES AGAINST GUCY2C AND THEIR APPLICATION

<130>PC72377

<160> 445

<170> Patent In version 3.5

<210> 1

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 1

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr Thr Ser Asp His Asn Pro

50 55 60

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr Val Leu Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 2

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 2

Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr

1 5 10

<210> 3

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 3

Phe Ile Arg Asn Arg Ala Arg Gly Tyr Thr Ser Asp His Asn Pro Ser

1 5 10 15

Val Lys Gly

<210> 4

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 4

Asp Arg Pro Ser Tyr Tyr Val Leu Asp Tyr

1 5 10

<210> 5

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 5

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser

20 25

<210> 6

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 6

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

1 5 10

<210> 7

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 7

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln

1 5 10 15

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

20 25 30

<210> 8

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 8

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

1 5 10

<210> 9

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 9

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr Thr Ser Asp His Asn Pro

50 55 60

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr Val Leu Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 10

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 10

Phe Ile Arg Asn Gln Ala Arg Gly Tyr Thr Ser Asp His Asn Pro Ser

1 5 10 15

Val Lys Gly

<210> 11

<211> 114

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 11

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Asn Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Tyr Pro Gly Asp Gly Met Thr Thr Tyr Thr Gln Lys Phe

50 55 60

Lys Asp Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Val Arg Lys Gly Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val

100 105 110

Ser Ser

<210> 12

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 12

Gly Tyr Thr Phe Thr Thr Tyr Trp Met Gln

1 5 10

<210> 13

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 13

Ala Ile Tyr Pro Gly Asp Gly Met Thr Thr Tyr Thr Gln Lys Phe Lys

1 5 10 15

Asp

<210> 14

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 14

Lys Gly Met Asp Tyr

15

<210> 15

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 15

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Asn Leu Ser Cys Lys Ala Ser

20 25

<210> 16

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 16

Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly

1 5 10

<210> 17

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 17

Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln

1 5 10 15

Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys Val Arg

20 25 30

<210> 18

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 18

Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser

1 5 10

<210> 19

<211> 120

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 19

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Lys Tyr

20 25 30

Trp Met Gln Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Tyr Pro Gly Asp Gly Phe Thr Thr Tyr Thr Gln Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asn Tyr Gly Arg Thr Tyr Gly Gly Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 20

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 20

Gly Tyr Thr Phe Thr Lys Tyr Trp Met Gln

1 5 10

<210> 21

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 21

Ala Ile Tyr Pro Gly Asp Gly Phe Thr Thr Tyr Thr Gln Lys Phe Lys

1 5 10 15

Gly

<210> 22

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 22

Arg Asn Tyr Gly Arg Thr Tyr Gly Gly Asp Tyr

1 5 10

<210> 23

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 23

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser

20 25

<210> 24

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 24

Trp Ile Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly

1 5 10

<210> 25

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 25

Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Asn Thr Ala Tyr Met Gln

1 5 10 15

Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg

20 25 30

<210> 26

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 26

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Lys Ser Ala Thr Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ala Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Ala Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 27

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 27

Gly Tyr Thr Phe Thr Ser Tyr Trp Met His

1 5 10

<210> 28

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 28

Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe Lys

1 5 10 15

Asn

<210> 29

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 29

Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val

1 5 10

<210> 30

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 30

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser

20 25

<210> 31

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 31

Lys Ala Thr Leu Thr Val Asp Lys Ser Ala Thr Thr Ala Tyr Met Gln

1 5 10 15

Leu Ser Ser Leu Thr Ala Glu Asp Ser Ala Val Tyr Tyr Cys Thr Arg

20 25 30

<210> 32

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 32

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser

1 5 10

<210> 33

<211> 115

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 33

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Thr Ala Ser Gly Phe Asn Ile Lys Asp Thr

20 25 30

Tyr Ile His Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile

35 40 45

Gly Arg Ile Asp Pro Ala Asn Gly Asn Ala Asn Tyr Asp Pro Lys Phe

50 55 60

Gln Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr

65 70 75 80

Leu Gln Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Phe Tyr Cys

85 90 95

Ser Ser Leu Gly Thr Gly Thr Tyr Trp Gly Gln Gly Thr Thr Leu Thr

100 105 110

Val Ser Ser

115

<210> 34

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 34

Gly Phe Asn Ile Lys Asp Thr Tyr Ile His

1 5 10

<210> 35

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 35

Arg Ile Asp Pro Ala Asn Gly Asn Ala Asn Tyr Asp Pro Lys Phe Gln

1 5 10 15

Gly

<210> 36

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 36

Leu Gly Thr Gly Thr Tyr

15

<210> 37

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 37

Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Thr Ala Ser

20 25

<210> 38

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 38

Trp Val Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile Gly

1 5 10

<210> 39

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 39

Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr Leu Gln

1 5 10 15

Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Phe Tyr Cys Ser Ser

20 25 30

<210> 40

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 40

Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser

1 5 10

<210> 41

<211> 123

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 41

Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr

20 25 30

Ile Met Leu Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile

35 40 45

Gly Asn Ser Asn Pro Tyr Tyr Gly Ser Thr Ser Tyr Asn Leu Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met His Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Gly Tyr Tyr Gly Ser Ser Pro Tyr Trp Tyr Phe Asp Val

100 105 110

Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 42

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 42

Gly Tyr Ser Phe Thr Asp Tyr Ile Met Leu

1 5 10

<210> 43

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 43

Asn Ser Asn Pro Tyr Tyr Gly Ser Thr Ser Tyr Asn Leu Lys Phe Lys

1 5 10 15

Gly

<210> 44

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 44

Ser Gly Tyr Tyr Gly Ser Ser Pro Tyr Trp Tyr Phe Asp Val

1 5 10

<210> 45

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 45

Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser

20 25

<210> 46

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 46

Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile Gly

1 5 10

<210> 47

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 47

Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr Met His

1 5 10 15

Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg

20 25 30

<210> 48

<211> 114

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 48

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Tyr Pro Gly Asp Gly Met Thr Thr Tyr Thr Gln Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Ala Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Arg Lys Gly Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

100 105 110

Ser Ser

<210> 49

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 49

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

1 5 10

<210> 50

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 50

Arg Phe Thr Ile Ser Ala Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln

1 5 10 15

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg

20 25 30

<210> 51

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 51

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 52

<211> 114

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 52

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met Gln Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Tyr Pro Gly Asp Gly Met Thr Thr Tyr Thr Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Arg Lys Gly Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

100 105 110

Ser Ser

<210> 53

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 53

Ala Ile Tyr Pro Gly Asp Gly Met Thr Thr Tyr Thr Gln Lys Phe Gln

1 5 10 15

Gly

<210> 54

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 54

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser

20 25

<210> 55

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 55

Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly

1 5 10

<210> 56

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 56

Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr Met Glu

1 5 10 15

Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg

20 25 30

<210> 57

<211> 123

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 57

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ser Phe Thr Asp Tyr

20 25 30

Ile Met Leu Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ser Asn Pro Tyr Tyr Gly Ser Thr Ser Tyr Asn Leu Lys Phe

50 55 60

Lys Gly Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Gly Tyr Tyr Gly Ser Ser Pro Tyr Trp Tyr Phe Asp Val

100 105 110

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 58

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 58

Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln

1 5 10 15

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

20 25 30

<210> 59

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 59

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

1 5 10

<210> 60

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 60

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 61

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 61

Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln

1 5 10 15

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg

20 25 30

<210> 62

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 62

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Ala Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 63

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 63

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Ala

1 5 10

<210> 64

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 64

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 65

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 65

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Ile His Pro Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 66

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 66

Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Ile His Pro Lys Phe Lys

1 5 10 15

Asn

<210> 67

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 67

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Asn Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 68

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 68

Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Asn Glu Lys Phe Lys

1 5 10 15

Asn

<210> 69

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 69

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 70

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 70

Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Val His Glu Lys Phe Lys

1 5 10 15

Asn

<210> 71

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 71

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ala Glu Gln Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 72

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 72

Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ala Glu Gln Phe Lys

1 5 10 15

Asn

<210> 73

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 73

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 74

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 74

Gly Phe Thr Phe Ser Ser Tyr Trp Met His

1 5 10

<210> 75

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 75

Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe Lys

1 5 10 15

Asp

<210> 76

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 76

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 77

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 77

Thr Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Arg Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 78

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 78

Trp Ala Ser Thr Arg Glu Ser

15

<210> 79

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 79

Lys Gln Ser Tyr Asp Leu Phe Thr

15

<210> 80

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 80

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys

20

<210> 81

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 81

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 82

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 82

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys

20 25 30

<210> 83

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 83

Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 84

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 84

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 85

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 85

Lys Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Arg Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 86

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 86

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys

20

<210> 87

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 87

Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Ser

1 5 10 15

<210> 88

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 88

Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys

20 25 30

<210> 89

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 89

Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 90

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 90

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 91

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 91

Thr Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Gln Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 92

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 92

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Leu Glu Glu Asp Asp Thr Ala Met Phe Phe Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 93

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 93

Arg Ala Ser Glu Ser Val Asp Asn Tyr Gly Ile Ser Phe Met Asn

1 5 10 15

<210> 94

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 94

Ala Ala Ser Asn Pro Gly Ser

15

<210> 95

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 95

Gln Gln Ser Lys Glu Val Pro Tyr Thr

15

<210> 96

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 96

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys

20

<210> 97

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 97

Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 98

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 98

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser

1 5 10 15

Leu Asn Ile His Pro Leu Glu Glu Asp Asp Thr Ala Met Phe Phe Cys

20 25 30

<210> 99

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 99

Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 100

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 100

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Phe

20 25 30

Asp Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Gln Gly Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Met Glu Glu Asp Asp Thr Ala Met Tyr Phe Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 101

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 101

Arg Ala Ser Glu Ser Val Asp Asn Phe Asp Ile Ser Phe Met Asn

1 5 10 15

<210> 102

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 102

Ala Ala Ser Asn Gln Gly Ser

15

<210> 103

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 103

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser

1 5 10 15

Leu Asn Ile His Pro Met Glu Glu Asp Asp Thr Ala Met Tyr Phe Cys

20 25 30

<210> 104

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 104

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Gly Glu Ser Val Asp Asn Phe

20 25 30

Asp Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Gln Gly Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Met Glu Glu Asp Asp Thr Ala Met Tyr Phe Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 105

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 105

Arg Ala Gly Glu Ser Val Asp Asn Phe Asp Ile Ser Phe Met Asn

1 5 10 15

<210> 106

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 106

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 107

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 107

Arg Ala Ser Glu Ser Val Asp Tyr Tyr Gly Thr Ser Leu Met Gln

1 5 10 15

<210> 108

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 108

Ala Ala Ser Asn Val Glu Ser

15

<210> 109

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 109

Gln Gln Thr Arg Lys Val Tyr Thr

15

<210> 110

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 110

Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 111

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 111

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser

1 5 10 15

Leu Asn Ile His Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys

20 25 30

<210> 112

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic construction

<400> 112

Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys Gly Val Thr Thr Ser

20 25 30

Gly Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg

85 90 95

Glu Phe Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105 110

<210> 113

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 113

Arg Ala Ser Lys Gly Val Thr Thr Ser Gly Tyr Ser Tyr Met His

1 5 10 15

<210> 114

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 114

Leu Ala Ser Asn Leu Glu Ser

15

<210> 115

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 115

Gln His Ser Arg Glu Phe Pro Leu Thr

15

<210> 116

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 116

Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys

20

<210> 117

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 117

Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Asn Ile His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys

20 25 30

<210> 118

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 118

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

1 5 10

<210> 119

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 119

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Val Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Arg Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser

65 70 75 80

Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Tyr Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 120

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 120

Lys Ser Ser Gln Ser Leu Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 121

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 121

Gln Gln Tyr Tyr Ser Tyr Pro Thr

15

<210> 122

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 122

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Val Ser Cys

20

<210> 123

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 123

Trp Tyr Gln Gln Arg Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 124

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 124

Gly Val Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Ser Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys

20 25 30

<210> 125

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 125

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 126

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 126

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys

20

<210> 127

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 127

Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 128

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 128

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 129

<211> 111

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 129

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 130

<211> 23

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 130

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys

20

<210> 131

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 131

Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr

1 5 10 15

<210> 132

<211> 32

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 132

Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys

20 25 30

<210> 133

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 133

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 134

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 134

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Tyr Pro Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 135

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 135

Trp Tyr Gln Gln Lys Pro Gly Lys Ser Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 136

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 136

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 137

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 137

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 138

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 138

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 139

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 139

Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 140

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 140

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Arg Tyr Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 141

<211> 7

<212> PROTEIN

<213> Artificial Sequene

<220>

<223> Synthetic peptide sequence

<400> 141

Ala Ala Ser Lys Arg Tyr Ser

15

<210> 142

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic construction

<400> 142

Gln Gln Thr Arg Lys Ala Tyr Thr

15

<210> 143

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 143

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Trp Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 144

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 144

Ala Ala Ser Lys Leu Trp Ser

15

<210> 145

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 145

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Val Ala Pro Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 146

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 146

Ala Ala Ser Lys Val Ala Pro

15

<210> 147

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 147

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 148

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 148

Arg Ala Ser Glu Ser Val Asp Tyr Tyr Gly Ser Ser Leu Leu Gln

1 5 10 15

<210> 149

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 149

Ala Ala Ser Lys Leu Ala Ser

15

<210> 150

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 150

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Ile Ala Pro Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 151

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 151

Ala Ala Ser Asn Ile Ala Pro

15

<210> 152

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 152

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Arg Tyr Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Glu Tyr Thr Phe Gly Gln Gly Thr Lys Leu Lys Ile Lys

100 105 110

<210> 153

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 153

Arg Ala Ser Glu Ser Val Asp Tyr Tyr Gly Ser Ser Leu Met Gln

1 5 10 15

<210> 154

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 154

Gln Gln Thr Arg Lys Glu Tyr Thr

15

<210> 155

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 155

Phe Gly Gln Gly Thr Lys Leu Lys Ile Lys

1 5 10

<210> 156

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 156

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Ala Pro Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 157

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 157

Ala Ala Ser Asn Val Ala Pro

15

<210> 158

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 158

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser His Arg Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 159

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic construction

<400> 159

Ala Ala Ser His Arg Ala Ser

15

<210> 160

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 160

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Glu Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 161

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 161

Ala Ala Ser Asn Val Ala Ser

15

<210> 162

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic construction

<400> 162

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly His Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Arg Tyr Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 163

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 163

Arg Ala Ser Glu Ser Val Asp Tyr Tyr Gly His Ser Leu Met Gln

1 5 10 15

<210> 164

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 164

Ala Ala Ser Asn Arg Tyr Ser

15

<210> 165

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 165

Gln Gln Thr Arg Lys Ala Tyr Ser

15

<210> 166

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 166

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Glu Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 167

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 167

Arg Ala Ser Gln Ser Val Asp Tyr Tyr Gly Ser Ser Leu Leu Gln

1 5 10 15

<210> 168

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 168

Asp Ala Ser Lys Leu Ala Ser

15

<210> 169

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 169

Gln Gln Glu Arg Lys Ala Tyr Thr

15

<210> 170

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 170

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Glu Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 171

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 171

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 172

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 172

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 173

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 173

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Glu Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 174

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 174

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Glu Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 175

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 175

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Asp Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 176

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 176

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Tyr Tyr Tyr Gly Met Asp Val

100 105

<210> 177

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 177

Gly Phe Thr Phe Ser Ser Tyr Trp Met Ser

1 5 10

<210> 178

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 178

Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val Lys

1 5 10 15

Gly

<210> 179

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 179

Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val

15

<210> 180

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 180

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 181

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 181

Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu Asn

1 5 10

<210> 182

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 182

Ala Ala Ser Ser Leu Gln Ser

15

<210> 183

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 183

Gln Gln Ser Tyr Ser Thr Pro Tyr Thr

15

<210> 184

<211> 107

<212> PROTEIN

<213> Homo sapiens

<400> 184

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 185

<211> 98

<212> PROTEIN

<213> Homo sapiens

<400> 185

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val

<210> 186

<211> 15

<212> PROTEIN

<213> Homo sapiens

<400> 186

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

1 5 10 15

<210> 187

<211> 217

<212> PROTEIN

<213> Homo sapiens

<400> 187

Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

100 105 110

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met

115 120 125

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

130 135 140

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

145 150 155 160

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

165 170 175

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

180 185 190

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

195 200 205

Lys Ser Leu Ser Leu Ser Pro Gly Lys

210 215

<210> 188

<211> 216

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 188

Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

100 105 110

Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Glu Glu Met

115 120 125

Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro

130 135 140

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

145 150 155 160

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

165 170 175

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

180 185 190

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

195 200 205

Lys Ser Leu Ser Leu Ser Pro Gly

210 215

<210> 189

<211> 216

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 189

Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

100 105 110

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met

115 120 125

Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro

130 135 140

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

145 150 155 160

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

165 170 175

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

180 185 190

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

195 200 205

Lys Ser Leu Ser Leu Ser Pro Gly

210 215

<210> 190

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 190

Gly Gly Gly Ser Gly Gly Gly Gly

15

<210> 191

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 191

Gly Gly Gly Gly Ser Gly Gly Gly Gly

15

<210> 192

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 192

Gly Cys Pro Pro Cys Pro

15

<210> 193

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 193

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

1 5 10 15

<210> 194

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 194

Arg Thr Ser Asp Gln

15

<210> 195

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 195

Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

1 5 10 15

<210> 196

<211> 462

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 196

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Leu Glu Glu Asp Asp Thr Ala Met Phe Phe Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly

115 120 125

Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser

130 135 140

Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro

145 150 155 160

Gly Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly

165 170 175

Tyr Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser

180 185 190

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr

210 215 220

Tyr Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

225 230 235 240

Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val

245 250 255

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

260 265 270

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

275 280 285

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

290 295 300

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

305 310 315 320

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

325 330 335

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

340 345 350

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro

355 360 365

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu

370 375 380

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

385 390 395 400

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

405 410 415

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

420 425 430

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

435 440 445

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 197

<211> 456

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 197

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Gln Gln Ser Gly

115 120 125

Ala Glu Leu Ala Arg Pro Gly Ala Ser Val Asn Leu Ser Cys Lys Ala

130 135 140

Ser Gly Tyr Thr Phe Thr Thr Tyr Trp Met Gln Trp Val Lys Gln Arg

145 150 155 160

Pro Gly Gln Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asp Gly

165 170 175

Met Thr Thr Tyr Thr Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr Ala

180 185 190

Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Ala Ser

195 200 205

Glu Asp Ser Ala Val Tyr Tyr Cys Val Arg Lys Gly Met Asp Tyr Trp

210 215 220

Gly Gln Gly Thr Ser Val Thr Val Ser Ser Gly Cys Pro Pro Cys Pro

225 230 235 240

Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

245 250 255

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

260 265 270

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

275 280 285

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

290 295 300

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

305 310 315 320

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

325 330 335

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

340 345 350

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met

355 360 365

Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro

370 375 380

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

385 390 395 400

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

405 410 415

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

420 425 430

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

435 440 445

Lys Ser Leu Ser Leu Ser Pro Gly

450 455

<210> 198

<211> 918

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 198

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Leu Glu Glu Asp Asp Thr Ala Met Phe Phe Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly

115 120 125

Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser

130 135 140

Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro

145 150 155 160

Gly Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly

165 170 175

Tyr Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser

180 185 190

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr

210 215 220

Tyr Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

225 230 235 240

Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val

245 250 255

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

260 265 270

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

275 280 285

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

290 295 300

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

305 310 315 320

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

325 330 335

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

340 345 350

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro

355 360 365

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu

370 375 380

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

385 390 395 400

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

405 410 415

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

420 425 430

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

435 440 445

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile

450 455 460

Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg

465 470 475 480

Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val Arg Ser

485 490 495

Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

500 505 510

Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp

515 520 525

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

530 535 540

Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln Ser Tyr

545 550 555 560

Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Gln Gln Ser Gly Ala Glu

580 585 590

Leu Ala Arg Pro Gly Ala Ser Val Asn Leu Ser Cys Lys Ala Ser Gly

595 600 605

Tyr Thr Phe Thr Thr Tyr Trp Met Gln Trp Val Lys Gln Arg Pro Gly

610 615 620

Gln Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asp Gly Met Thr

625 630 635 640

Thr Tyr Thr Gln Lys Phe Lys Asp Lys Ala Thr Leu Thr Ala Asp Lys

645 650 655

Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser Leu Ala Ser Glu Asp

660 665 670

Ser Ala Val Tyr Tyr Cys Val Arg Lys Gly Met Asp Tyr Trp Gly Gln

675 680 685

Gly Thr Ser Val Thr Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro

690 695 700

Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

705 710 715 720

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

725 730 735

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

740 745 750

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

755 760 765

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

770 775 780

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

785 790 795 800

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

805 810 815

Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys

820 825 830

Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp

835 840 845

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

850 855 860

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser

865 870 875 880

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

885 890 895

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

900 905 910

Leu Ser Leu Ser Pro Gly

915

<210> 199

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 199

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 200

<211> 464

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 200

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Gln Val Gln Leu Gln Gln Pro Gly

115 120 125

Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala

130 135 140

Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val Lys Gln Arg

145 150 155 160

Pro Gly Gln Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly

165 170 175

Leu Thr Asn Tyr Ile Glu Lys Phe Lys Asn Lys Ala Thr Leu Thr Val

180 185 190

Asp Lys Ser Ala Thr Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ala

195 200 205

Glu Asp Ser Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu

210 215 220

Gly Tyr Trp Phe Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val

225 230 235 240

Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro

245 250 255

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

260 265 270

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

275 280 285

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

290 295 300

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

305 310 315 320

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

325 330 335

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

340 345 350

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

355 360 365

Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser

370 375 380

Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

385 390 395 400

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

405 410 415

Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys

420 425 430

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

435 440 445

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 201

<211> 925

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 201

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile Val

450 455 460

Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg Ala

465 470 475 480

Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Arg

485 490 495

Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys

500 505 510

Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg

515 520 525

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser

530 535 540

Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln Ser Tyr Asp

545 550 555 560

Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly

565 570 575

Ser Gly Gly Gly Gly Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu

580 585 590

Val Lys Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr

595 600 605

Thr Phe Thr Ser Tyr Trp Met His Trp Val Lys Gln Arg Pro Gly Gln

610 615 620

Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn

625 630 635 640

Tyr Ile Glu Lys Phe Lys Asn Lys Ala Thr Leu Thr Val Asp Lys Ser

645 650 655

Ala Thr Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ala Glu Asp Ser

660 665 670

Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp

675 680 685

Phe Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly

690 695 700

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

705 710 715 720

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

725 730 735

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

740 745 750

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

755 760 765

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

770 775 780

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

785 790 795 800

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

805 810 815

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

820 825 830

Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val

835 840 845

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

850 855 860

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

865 870 875 880

Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

885 890 895

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

900 905 910

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

915 920 925

<210> 202

<211> 463

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 202

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Val Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Arg Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser

65 70 75 80

Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Tyr Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly

115 120 125

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

130 135 140

Ser Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala

145 150 155 160

Pro Gly Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg

165 170 175

Gly Tyr Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile

180 185 190

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

195 200 205

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser

210 215 220

Tyr Tyr Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

225 230 235 240

Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser

245 250 255

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

260 265 270

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

275 280 285

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

290 295 300

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

305 310 315 320

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

325 330 335

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

340 345 350

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu

355 360 365

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys

370 375 380

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

385 390 395 400

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

405 410 415

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

420 425 430

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

435 440 445

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 203

<211> 465

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 203

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Gln Gln Ser Gly

115 120 125

Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala

130 135 140

Ser Gly Tyr Ser Phe Thr Asp Tyr Ile Met Leu Trp Val Lys Gln Ser

145 150 155 160

His Gly Lys Ser Leu Glu Trp Ile Gly Asn Ser Asn Pro Tyr Tyr Gly

165 170 175

Ser Thr Ser Tyr Asn Leu Lys Phe Lys Gly Lys Ala Thr Leu Thr Val

180 185 190

Asp Lys Ser Ser Ser Thr Ala Tyr Met His Leu Asn Ser Leu Thr Ser

195 200 205

Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Gly Tyr Tyr Gly Ser

210 215 220

Ser Pro Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr

225 230 235 240

Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

370 375 380

Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly

465

<210> 204

<211> 928

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 204

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Val Gly

1 5 10 15

Glu Lys Val Thr Val Ser Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser

20 25 30

Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Arg Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser

65 70 75 80

Ile Ser Ser Val Lys Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln

85 90 95

Tyr Tyr Ser Tyr Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly

115 120 125

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

130 135 140

Ser Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala

145 150 155 160

Pro Gly Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg

165 170 175

Gly Tyr Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile

180 185 190

Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu

195 200 205

Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser

210 215 220

Tyr Tyr Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

225 230 235 240

Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser

245 250 255

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

260 265 270

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

275 280 285

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

290 295 300

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

305 310 315 320

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

325 330 335

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

340 345 350

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu

355 360 365

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys

370 375 380

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

385 390 395 400

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

405 410 415

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

420 425 430

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

435 440 445

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp

450 455 460

Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu

465 470 475 480

Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val Arg

485 490 495

Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro

500 505 510

Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val Pro

515 520 525

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile

530 535 540

Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln Ser

545 550 555 560

Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Gly

565 570 575

Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Gln Gln Ser Gly Pro

580 585 590

Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Ala Ser

595 600 605

Gly Tyr Ser Phe Thr Asp Tyr Ile Met Leu Trp Val Lys Gln Ser His

610 615 620

Gly Lys Ser Leu Glu Trp Ile Gly Asn Ser Asn Pro Tyr Tyr Gly Ser

625 630 635 640

Thr Ser Tyr Asn Leu Lys Phe Lys Gly Lys Ala Thr Leu Thr Val Asp

645 650 655

Lys Ser Ser Ser Thr Ala Tyr Met His Leu Asn Ser Leu Thr Ser Glu

660 665 670

Asp Ser Ala Val Tyr Tyr Cys Ala Arg Ser Gly Tyr Tyr Gly Ser Ser

675 680 685

Pro Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val

690 695 700

Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro

705 710 715 720

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

725 730 735

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

740 745 750

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

755 760 765

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

770 775 780

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

785 790 795 800

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

805 810 815

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

820 825 830

Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser

835 840 845

Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

850 855 860

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

865 870 875 880

Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys

885 890 895

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

900 905 910

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

915 920 925

<210> 205

<211> 462

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 205

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly

115 120 125

Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser

130 135 140

Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro

145 150 155 160

Gly Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly

165 170 175

Tyr Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser

180 185 190

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr

210 215 220

Tyr Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

225 230 235 240

Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val

245 250 255

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

260 265 270

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

275 280 285

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

290 295 300

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

305 310 315 320

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

325 330 335

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

340 345 350

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro

355 360 365

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu

370 375 380

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

385 390 395 400

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

405 410 415

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

420 425 430

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

435 440 445

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 206

<211> 456

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 206

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly

115 120 125

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

130 135 140

Ser Gly Tyr Thr Phe Thr Thr Tyr Trp Met Gln Trp Val Arg Gln Ala

145 150 155 160

Pro Gly Lys Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asp Gly

165 170 175

Met Thr Thr Tyr Thr Gln Lys Phe Lys Asp Arg Phe Thr Ile Ser Ala

180 185 190

Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Val Arg Lys Gly Met Asp Tyr Trp

210 215 220

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Cys Pro Pro Cys Pro

225 230 235 240

Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys

245 250 255

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

260 265 270

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

275 280 285

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

290 295 300

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

305 310 315 320

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

325 330 335

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

340 345 350

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met

355 360 365

Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro

370 375 380

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

385 390 395 400

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

405 410 415

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

420 425 430

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

435 440 445

Lys Ser Leu Ser Leu Ser Pro Gly

450 455

<210> 207

<211> 918

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 207

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr

20 25 30

Gly Ile Ser Phe Met Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Pro Gly Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly

100 105 110

Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly

115 120 125

Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser

130 135 140

Gly Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro

145 150 155 160

Gly Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly

165 170 175

Tyr Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser

180 185 190

Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr

210 215 220

Tyr Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

225 230 235 240

Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val

245 250 255

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

260 265 270

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

275 280 285

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

290 295 300

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

305 310 315 320

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

325 330 335

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

340 345 350

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro

355 360 365

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu

370 375 380

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

385 390 395 400

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

405 410 415

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

420 425 430

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

435 440 445

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile

450 455 460

Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg

465 470 475 480

Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val Arg Ser

485 490 495

Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

500 505 510

Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp

515 520 525

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

530 535 540

Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln Ser Tyr

545 550 555 560

Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

580 585 590

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

595 600 605

Tyr Thr Phe Thr Thr Tyr Trp Met Gln Trp Val Arg Gln Ala Pro Gly

610 615 620

Lys Gly Leu Glu Trp Ile Gly Ala Ile Tyr Pro Gly Asp Gly Met Thr

625 630 635 640

Thr Tyr Thr Gln Lys Phe Lys Asp Arg Phe Thr Ile Ser Ala Asp Lys

645 650 655

Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

660 665 670

Thr Ala Val Tyr Tyr Cys Val Arg Lys Gly Met Asp Tyr Trp Gly Gln

675 680 685

Gly Thr Leu Val Thr Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro

690 695 700

Glu Ala Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

705 710 715 720

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

725 730 735

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

740 745 750

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

755 760 765

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

770 775 780

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

785 790 795 800

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

805 810 815

Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys

820 825 830

Asn Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp

835 840 845

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

850 855 860

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser

865 870 875 880

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

885 890 895

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

900 905 910

Leu Ser Leu Ser Pro Gly

915

<210> 208

<211> 217

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 208

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr

100 105 110

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 209

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 209

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

225 230 235 240

Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly

450

<210> 210

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 210

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 211

<211> 464

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 211

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly

115 120 125

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

130 135 140

Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala

145 150 155 160

Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly

165 170 175

Leu Thr Asn Tyr Ile Glu Lys Phe Lys Asn Arg Phe Thr Ile Ser Val

180 185 190

Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu

210 215 220

Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val

225 230 235 240

Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro

245 250 255

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

260 265 270

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

275 280 285

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

290 295 300

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

305 310 315 320

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

325 330 335

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

340 345 350

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

355 360 365

Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser

370 375 380

Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

385 390 395 400

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

405 410 415

Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys

420 425 430

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

435 440 445

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 212

<211> 925

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 212

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile Val

450 455 460

Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly Glu Arg Ala

465 470 475 480

Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Arg

485 490 495

Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys

500 505 510

Leu Leu Ile Ser Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Asp Arg

515 520 525

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser

530 535 540

Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Lys Gln Ser Tyr Asp

545 550 555 560

Leu Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly

565 570 575

Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

580 585 590

Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr

595 600 605

Thr Phe Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Lys

610 615 620

Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn

625 630 635 640

Tyr Ile Glu Lys Phe Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala

645 650 655

Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

660 665 670

Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp

675 680 685

Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly

690 695 700

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

705 710 715 720

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

725 730 735

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

740 745 750

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

755 760 765

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

770 775 780

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

785 790 795 800

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

805 810 815

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

820 825 830

Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val

835 840 845

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

850 855 860

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

865 870 875 880

Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

885 890 895

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

900 905 910

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

915 920 925

<210> 213

<211> 483

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 213

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Asp Ile Gln Leu Thr Gln

130 135 140

Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

145 150 155 160

Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr Gly Ser Ser Leu Leu Gln

165 170 175

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala

180 185 190

Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

195 200 205

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

210 215 220

Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg Lys Ala Tyr Thr Phe Gly

225 230 235 240

Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Ser Asp Gln Glu Pro Lys

245 250 255

Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala

260 265 270

Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

275 280 285

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

290 295 300

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

305 310 315 320

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

325 330 335

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

340 345 350

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

355 360 365

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

370 375 380

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

385 390 395 400

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

405 410 415

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

420 425 430

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

435 440 445

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

450 455 460

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

465 470 475 480

Leu Ser Pro

<210> 214

<211> 217

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 214

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr

100 105 110

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 215

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 215

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

225 230 235 240

Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly

450

<210> 216

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 216

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 217

<211> 465

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 217

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser

115 120 125

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

130 135 140

Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val Arg Gln

145 150 155 160

Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn

165 170 175

Gly Leu Thr Asn Val His Glu Lys Phe Lys Asn Arg Phe Thr Ile Ser

180 185 190

Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr

210 215 220

Glu Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr

225 230 235 240

Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

370 375 380

Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly

465

<210> 218

<211> 926

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 218

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile Gln

450 455 460

Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val

465 470 475 480

Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Gln

485 490 495

Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys

500 505 510

Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Ser Arg

515 520 525

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser

530 535 540

Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Ser Tyr Asp

545 550 555 560

Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

580 585 590

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

595 600 605

Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly

610 615 620

Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr

625 630 635 640

Asn Val His Glu Lys Phe Lys Asn Arg Phe Thr Ile Ser Val Asp Lys

645 650 655

Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

660 665 670

Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr

675 680 685

Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

690 695 700

Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val

705 710 715 720

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

725 730 735

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

740 745 750

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

755 760 765

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

770 775 780

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

785 790 795 800

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

805 810 815

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

820 825 830

Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala

835 840 845

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

850 855 860

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

865 870 875 880

Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg

885 890 895

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

900 905 910

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

915 920 925

<210> 219

<211> 10

<212> PROTEIN

<213> Homo sapiens

<400> 219

Glu Pro Lys Ser Ser Asp Lys Thr His Thr

1 5 10

<210> 220

<211> 465

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 220

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser

115 120 125

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

130 135 140

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln

145 150 155 160

Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn

165 170 175

Glu Leu Thr Asn Val His Glu Lys Phe Lys Asp Arg Phe Thr Ile Ser

180 185 190

Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr

210 215 220

Glu Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr

225 230 235 240

Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

370 375 380

Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly

465

<210> 221

<211> 926

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 221

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile Gln

450 455 460

Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val

465 470 475 480

Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Gln

485 490 495

Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys

500 505 510

Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Ser Arg

515 520 525

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser

530 535 540

Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Ser Tyr Asp

545 550 555 560

Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly

565 570 575

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

580 585 590

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

595 600 605

Phe Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly

610 615 620

Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr

625 630 635 640

Asn Val His Glu Lys Phe Lys Asp Arg Phe Thr Ile Ser Val Asp Lys

645 650 655

Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp

660 665 670

Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr

675 680 685

Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

690 695 700

Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val

705 710 715 720

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

725 730 735

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

740 745 750

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

755 760 765

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

770 775 780

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

785 790 795 800

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

805 810 815

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

820 825 830

Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala

835 840 845

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

850 855 860

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

865 870 875 880

Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg

885 890 895

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

900 905 910

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

915 920 925

<210> 222

<211> 5

<212> PROTEIN

<213> Homo sapiens

<400> 222

Cys Pro Pro Cys Pro

15

<210> 223

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 223

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

225 230 235 240

Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly

450

<210> 224

<211> 1080

<212> PROTEIN

<213> Homo sapiens

<400> 224

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val Leu

85 90 95

Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Thr

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Ile

210 215 220

Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Leu Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Gly Arg Gly Pro Gln Ile Leu Met Ile Ala Val Phe Thr Leu

405 410 415

Thr Gly Ala Val Val Leu Leu Leu Leu Val Ala Leu Leu Met Leu Arg

420 425 430

Lys Tyr Arg Lys Asp Tyr Glu Leu Arg Gln Lys Lys Trp Ser His Ile

435 440 445

Pro Pro Glu Asn Ile Phe Pro Leu Glu Thr Asn Glu Thr Asn His Val

450 455 460

Ser Leu Lys Ile Asp Asp Asp Lys Arg Arg Asp Thr Ile Gln Arg Leu

465 470 475 480

Arg Gln Cys Lys Tyr Asp Lys Lys Arg Val Ile Leu Lys Asp Leu Lys

485 490 495

His Asn Asp Gly Asn Phe Thr Glu Lys Gln Lys Ile Glu Leu Asn Lys

500 505 510

Leu Leu Gln Ile Asp Tyr Tyr Asn Leu Thr Lys Phe Tyr Gly Thr Val

515 520 525

Lys Leu Asp Thr Met Ile Phe Gly Val Ile Glu Tyr Cys Glu Arg Gly

530 535 540

Ser Leu Arg Glu Val Leu Asn Asp Thr Ile Ser Tyr Pro Asp Gly Thr

545 550 555 560

Phe Met Asp Trp Glu Phe Lys Ile Ser Val Leu Tyr Asp Ile Ala Lys

565 570 575

Gly Met Ser Tyr Leu His Ser Ser Lys Thr Glu Val His Gly Arg Leu

580 585 590

Lys Ser Thr Asn Cys Val Val Asp Ser Arg Met Val Val Lys Ile Thr

595 600 605

Asp Phe Gly Cys Asn Ser Ile Leu Pro Pro Lys Lys Asp Leu Trp Thr

610 615 620

Ala Pro Glu His Leu Arg Gln Ala Asn Ile Ser Gln Lys Gly Asp Val

625 630 635 640

Tyr Ser Tyr Gly Ile Ile Ala Gln Glu Ile Ile Leu Arg Lys Glu Thr

645 650 655

Phe Tyr Thr Leu Ser Cys Arg Asp Arg Asn Glu Lys Ile Phe Arg Val

660 665 670

Glu Asn Ser Asn Gly Met Lys Pro Phe Arg Pro Asp Leu Phe Leu Glu

675 680 685

Thr Ala Glu Glu Lys Glu Leu Glu Val Tyr Leu Leu Val Lys Asn Cys

690 695 700

Trp Glu Glu Asp Pro Glu Lys Arg Pro Asp Phe Lys Lys Ile Glu Thr

705 710 715 720

Thr Leu Ala Lys Ile Phe Gly Leu Phe His Asp Gln Lys Asn Glu Ser

725 730 735

Tyr Met Asp Thr Leu Ile Arg Arg Leu Gln Leu Tyr Ser Arg Asn Leu

740 745 750

Glu His Leu Val Glu Glu Arg Thr Gln Leu Tyr Lys Ala Glu Arg Asp

755 760 765

Arg Ala Asp Arg Leu Asn Phe Met Leu Leu Pro Arg Leu Val Val Lys

770 775 780

Ser Leu Lys Glu Lys Gly Phe Val Glu Pro Glu Leu Tyr Glu Glu Val

785 790 795 800

Thr Ile Tyr Phe Ser Asp Ile Val Gly Phe Thr Thr Ile Cys Lys Tyr

805 810 815

Ser Thr Pro Met Glu Val Val Asp Met Leu Asn Asp Ile Tyr Lys Ser

820 825 830

Phe Asp His Ile Val Asp His His Asp Val Tyr Lys Val Glu Thr Ile

835 840 845

Gly Asp Ala Tyr Met Val Ala Ser Gly Leu Pro Lys Arg Asn Gly Asn

850 855 860

Arg His Ala Ile Asp Ile Ala Lys Met Ala Leu Glu Ile Leu Ser Phe

865 870 875 880

Met Gly Thr Phe Glu Leu Glu His Leu Pro Gly Leu Pro Ile Trp Ile

885 890 895

Arg Ile Gly Val His Ser Gly Pro Cys Ala Ala Gly Val Val Gly Ile

900 905 910

Lys Met Pro Arg Tyr Cys Leu Phe Gly Asp Thr Val Asn Thr Ala Ser

915 920 925

Arg Met Glu Ser Thr Gly Leu Pro Leu Arg Ile His Val Ser Gly Ser

930 935 940

Thr Ile Ala Ile Leu Lys Arg Thr Glu Cys Gln Phe Leu Tyr Glu Val

945 950 955 960

Arg Gly Glu Thr Tyr Leu Lys Gly Arg Gly Asn Glu Thr Thr Tyr Trp

965 970 975

Leu Thr Gly Met Lys Asp Gln Lys Phe Asn Leu Pro Thr Pro Pro Thr

980 985 990

Val Glu Asn Gln Gln Arg Leu Gln Ala Glu Phe Ser Asp Met Ile Ala

995 1000 1005

Asn Ser Leu Gln Lys Arg Gln Ala Ala Gly Ile Arg Ser Gln Lys

1010 1015 1020

Pro Arg Arg Val Ala Ser Tyr Lys Lys Gly Thr Leu Glu Tyr Leu

1025 1030 1035

Gln Leu Asn Thr Thr Asp Lys Glu Ser Thr Tyr Phe Thr Arg Thr

1040 1045 1050

Arg Pro Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Ala Ala

1055 1060 1065

Asn Asp Ile Leu Asp Tyr Lys Asp Asp Asp Asp Lys

1070 1075 1080

<210> 225

<211> 3240

<212> DNA

<213> Homo sapiens

<400> 225

tcccaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatgggc 60

aactcagcct ttgcagagcc cctgaaaaac ttggaagatg cggtgaatga ggggctggaa 120

atagtgagag gacgtctgca aaatgctggc ctaaatgtga ctgtgaacgc tactttcatg 180

tattcggatg gtctgattca taactcaggc gactgccgga gtagcacctg tgaaggcctc 240

gacctactca ggaaaatttc aaatgcacaa cggatgggct gtgtcctcat agggccctca 300

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 360

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 420

gctagaaagt tgatgtactt cttggttaac ttttggaaaa ccaacgatct gcccttcaaa 480

acttattcct ggagcacttc gtatgtttac aagaatggta cagaaactga ggactgtttc 540

tggtacctta atgctctgga ggctagcgtt tcctatttct cccacgaact cggctttaag 600

gtggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 660

aatgtgatta ttatgtgtgg tggtccagag ttcctctaca agctgaaggg tgaccgagca 720

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta cttggaggac 780

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgctgtc tcctgggaat 840

tcccttctaa atagctcttt ctccaggaat ctatcaccaa caaaacgaga ctttgctctt 900

gcctatttga atggaatcct gctctttgga catatgctga agatatttct tgaaaatgga 960

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1020

gacggtccag tgaccttgga tgactgggg gatgttgaca gtaccatggt gcttctgtat 1080

acctctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaataag 1140

acctatcctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1200

attacaggcc ggggccctca gatcctgatg attgcagtct tcaccctcac tggagctgtg 1260

gtgctgctcc tgctcgtcgc tctcctgatg ctcagaaaat atagaaaaga ttatgaactt 1320

cgtcagaaaa aatggtccca cattcctcct gaaaatatct ttcctctgga gaccaatgag 1380

accaatcatg ttagcctcaa gatcgatgat gacaaaagac gagatacaat ccagagacta 1440

cgacagtgca aatacgacaa aaagcgagtg attctcaaag atctcaagca caatgatggt 1500

aatttcactg aaaaacagaa gatagaattg aacaagttgc ttcagattga ctattacaac 1560

ctgaccaagt tctacggcac agtgaaactt gataccatga tcttcggggt gatagaatac 1620

tgtgagagag gatccctccg ggaagtttta aatgacacaa tttcctaccc tgatggcaca 1680

ttcatggatt gggagtttaa gatctctgtc ttgtatgaca ttgctaaggg aatgtcatat 1740

ctgcactcca gtaagacaga agtccatggt cgtctgaaat ctaccaactg cgtagtggac 1800

agtagaatgg tggtgaagat cactgatttt ggctgcaatt ccattttacc tccaaaaaag 1860

gacctgtgga cagctccaga gcacctccgc caagccaaca tctctcagaa aggagatgtg 1920

tacagctatg ggatcatcgc acaggagatc attctgcgga aagaaacctt ctacactttg 1980

agctgtcggg accggaatga gaagattttc agagtggaaa attccaatgg aatgaaaccc 2040

ttccgcccag atttattctt ggaaacagca gaggaaaaag agctagaagt gtacctactt 2100

gtaaaaaact gttgggagga agatccagaa aagagaccag atttcaaaaa aattgagact 2160

acacttgcca agatatttgg actttttcat gaccaaaaaa atgaaagcta tatggatacc 2220

ttgatccgac gtctacagct atattctcga aacctggaac atctggtaga ggaaaggaca 2280

cagctgtaca aggcagagag ggacagggct gacagactta actttatgtt gcttccaagg 2340

ctagtggtaa agtctctgaa ggagaaaggc tttgtggagc cggaactata tgaggaagtt 2400

acaatctact tcagtgacat tgtaggtttc actactatct gcaaatacag cacccccatg 2460

gaagtggtgg acatgcttaa tgacatctat aagagttttg accacattgt tgatcatcat 2520

gatgtctaca aggtggaaac catcggtgat gcgtacatgg tggctagtgg tttgcctaag 2580

agaaatggca atcggcatgc aatagacatt gccaagatgg ccttggaaat cctcagcttc 2640

atggggacct ttgagctgga gcatcttcct ggcctcccaa tatggattcg cattggagtt 2700

cactctggtc cctgtgctgc tggagttgtg ggaatcaaga tgcctcgtta ttgtctattt 2760

ggagatacgg tcaacacagc ctctaggatg gaatccactg gcctcccttt gagaattcac 2820

gtgagtggct ccaccatagc catcctgaag agaactgagt gccagttcct ttatgaagtg 2880

agaggagaaa catacttaaa gggaagagga aatgagacta cctactggct gactgggatg 2940

aaggaccaga aattcaacct gccaacccct cctactgtgg agaatcaaca gcgtttgcaa 3000

gcagaatttt cagacatgat tgccaactct ttacagaaaa gacaggcagc agggataaga 3060

agccaaaaac cgacgggt agccagctat aaaaaaggca ctctggaata cttgcagctg 3120

aataccacag acaaggag cacctatttt acgcgtacgc ggccgctcga gcagaaactc 3180

atctcagaag aggatctggc agcaaatgat atcctggatt acaaggatga cgacgataag 3240

<210> 226

<211> 1080

<212> PROTEIN

<213> Macaca fascicularis

<400> 226

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Asp Asn Ser Ala Phe Ala Glu Pro Leu Glu Asn Val Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Ser Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Lys Arg Met Gly Cys Val Leu

85 90 95

Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Thr Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Ser

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Ser Phe Lys Leu Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Val

210 215 220

Met Cys Gly Asp Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Gln Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Thr Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Gln Thr Asn Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Asp Arg Gly Pro Gln Ile Leu Met Ile Ala Val Phe Thr Leu

405 410 415

Thr Gly Ala Val Val Leu Leu Leu Leu Val Ala Leu Leu Met Leu Arg

420 425 430

Lys Tyr Lys Lys Asp Tyr Glu Leu Arg Gln Lys Lys Trp Ser His Ile

435 440 445

Pro Pro Glu Asn Ile Phe Pro Leu Glu Thr Asn Glu Thr Asn His Val

450 455 460

Ser Leu Lys Ile Asp Asp Asp Lys Arg Arg Asp Thr Ile Gln Arg Leu

465 470 475 480

Arg Gln Cys Lys Tyr Asp Lys Lys Arg Val Ile Leu Lys Asp Leu Lys

485 490 495

His Asn Asp Gly Asn Phe Thr Glu Lys Gln Lys Ile Glu Leu Asn Lys

500 505 510

Leu Leu Gln Ile Asp Tyr Tyr Asn Leu Thr Lys Phe Tyr Gly Thr Val

515 520 525

Lys Leu Asp Thr Met Ile Phe Gly Val Ile Glu Tyr Cys Glu Arg Gly

530 535 540

Ser Leu Arg Glu Val Leu Asn Asp Thr Ile Ser Tyr Pro Asp Gly Thr

545 550 555 560

Phe Met Asp Trp Glu Phe Lys Ile Ser Val Leu Tyr Asp Ile Ala Lys

565 570 575

Gly Met Ser Tyr Leu His Ser Ser Lys Thr Glu Val His Gly Arg Leu

580 585 590

Lys Ser Thr Asn Cys Val Val Asp Ser Arg Met Val Val Lys Ile Thr

595 600 605

Asp Phe Gly Cys Asn Ser Ile Leu Pro Pro Lys Lys Asp Leu Trp Thr

610 615 620

Ala Pro Glu His Leu Arg Gln Ala Asn Val Ser Gln Lys Gly Asp Val

625 630 635 640

Tyr Ser Tyr Gly Ile Ile Ala Gln Glu Ile Ile Leu Arg Lys Glu Thr

645 650 655

Phe Tyr Thr Ser Ser Cys Arg Asp Arg Asn Glu Lys Ile Phe Arg Val

660 665 670

Glu Asn Ser Asn Gly Met Lys Pro Phe Arg Pro Asp Leu Phe Leu Glu

675 680 685

Thr Ala Glu Glu Lys Glu Leu Glu Val Tyr Leu Leu Val Lys Ser Cys

690 695 700

Trp Glu Glu Asp Pro Glu Lys Arg Pro Asp Phe Lys Lys Ile Glu Thr

705 710 715 720

Thr Leu Ala Lys Ile Phe Gly Leu Phe His Asp Gln Lys Asn Glu Ser

725 730 735

Tyr Met Asp Thr Leu Ile Arg Arg Leu Gln Leu Tyr Ser Arg Asn Leu

740 745 750

Glu His Leu Val Glu Glu Arg Thr Gln Leu Tyr Lys Ala Glu Arg Asp

755 760 765

Arg Ala Asp Arg Leu Asn Phe Met Leu Leu Pro Arg Leu Val Val Lys

770 775 780

Ser Leu Lys Glu Lys Gly Phe Val Glu Pro Glu Leu Tyr Glu Glu Val

785 790 795 800

Thr Ile Tyr Phe Ser Asp Ile Val Gly Phe Thr Thr Ile Cys Lys Tyr

805 810 815

Ser Thr Pro Met Glu Val Val Asp Met Leu Asn Asp Ile Tyr Lys Ser

820 825 830

Phe Asp His Ile Val Asp His His Asp Val Tyr Lys Val Glu Thr Ile

835 840 845

Gly Asp Ala Tyr Met Val Ala Ser Gly Leu Pro Lys Arg Asn Gly Asn

850 855 860

Arg His Ala Ile Asp Ile Ala Lys Met Ala Leu Glu Ile Leu Ser Phe

865 870 875 880

Met Gly Thr Phe Glu Leu Glu His Leu Pro Gly Leu Pro Ile Trp Ile

885 890 895

Arg Ile Gly Val His Ser Gly Pro Cys Ala Ala Gly Val Val Gly Ile

900 905 910

Lys Met Pro Arg Tyr Cys Leu Phe Gly Asp Thr Val Asn Thr Ala Ser

915 920 925

Arg Met Glu Ser Thr Gly Leu Pro Leu Arg Ile His Val Ser Gly Ser

930 935 940

Thr Ile Ala Ile Leu Lys Arg Thr Glu Cys Gln Phe Leu Tyr Glu Val

945 950 955 960

Arg Gly Glu Thr Tyr Leu Lys Gly Arg Gly Asn Glu Thr Thr Tyr Trp

965 970 975

Leu Thr Gly Met Lys Asp Gln Lys Phe Asn Leu Pro Thr Pro Pro Thr

980 985 990

Val Glu Asn Gln Gln Arg Leu Gln Ala Glu Phe Ser Asp Met Ile Ala

995 1000 1005

Asn Ser Leu Gln Lys Arg Gln Ala Ala Gly Ile Arg Ser Gln Lys

1010 1015 1020

Pro Arg Arg Val Ala Ser Tyr Lys Lys Gly Thr Leu Glu Tyr Leu

1025 1030 1035

Gln Leu Asn Thr Thr Asp Lys Glu Ser Thr Tyr Phe Thr Arg Thr

1040 1045 1050

Arg Pro Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Ala Ala

1055 1060 1065

Asn Asp Ile Leu Asp Tyr Lys Asp Asp Asp Asp Lys

1070 1075 1080

<210> 227

<211> 3240

<212> DNA

<213> Macaca fascicularis

<400> 227

tcacaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatggac 60

aactcagcct ttgcagagcc cctggaaaac gtggaagatg cggtgaatga ggggctggaa 120

atagtgagag gacgtctgca aaacgctggc ctaaatgtga ctgtgaatgc ttctttcatg 180

tattcggatg gtctgattca taactccggc gactgccgga gcagcacctg tgaaggcctt 240

gacctactca ggaaaatttc aaatgcaaaa cggatgggct gtgtcctcat ggggccctca 300

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 360

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 420

gctagaaagt tgacatactt cttggttaac ttttggaaaa ccaatgatct acccttcaaa 480

acttattcct ggagcacttc gtatgtttac aagaatggta cggagtccga ggactgtttc 540

tggtacctta acgctctgga ggccagtgtt tcctatttct cccacgaact cagttttaag 600

ttggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 660

aatgtgattg ttatgtgtgg tgatccagag ttcctctaca agttgaaggg tgaccgagca 720

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta ctttgaggac 780

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgcagtc tcctggcaat 840

tccctcctaa atagctcttt ctccaggaat ctatccccaa caaaacgaga ctttgctctt 900

gcctatttga atggaatcct gctctttgga catatgctaa agacatttct tgaaaatgga 960

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1020

gacggtccag tgaccttgga tgactgggg gatgtggaca gtaccatggt gcttctgtat 1080

acgtctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaatcag 1140

accaaccctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1200

attacagacc ggggccctca gatcctgatg attgcagtct tcaccctcac cggagctgtg 1260

gtgctgctcc tgcttgtcgc tctcctgatg ctcagaaaat ataaaaaaga ttatgaactt 1320

cgtcagaaaa aatggtccca cattcctcct gaaaatatct ttcctctgga gaccaatgag 1380

accaatcacg ttagcctgaa gatcgatgat gacaaaagac gagatacaat cccagacta 1440

cgacagtgca aatacgacaa aaagcgagtg attctcaaag atctcaagca caatgatggt 1500

aatttcactg aaaaacagaa gatagaattg aacaagttgc ttcagattga ctattacaac 1560

ctgaccaagt tctatggcac cgtgaaactt gataccatga tcttcggggt gatagaatac 1620

tgtgagagag gatccctccg ggaagtttta aatgacacaa tttcctaccc tgatggcaca 1680

ttcatggatt gggagtttaa gatctctgtc ctgtatgaca ttgctaaggg aatgtcatat 1740

ctgcactcca gtaagacaga agtccatggt cgtctgaaat ctaccaactg cgtagtggac 1800

agtagaatgg tggtgaagat cactgatttt ggctgcaatt ccattttacc tccaaaaaaa 1860

gacctgtgga cagctccaga gcacctccgc caagccaacg tctctcagaa aggagatgtg 1920

tacagctacg ggatcatcgc acaggagatc atcctgcgga aagaaacctt ctacacttcg 1980

agctgtcgag accggaacga gaagattttc agagtggaaa attccaatgg aatgaaaccc 2040

ttccgtccag atttattctt ggaaacggca gaggaaaaag agctagaagt gtacctactt 2100

gtaaaaagct gttgggaaga agatccagaa aagagaccag atttcaaaaa aattgagact 2160

acacttgcca agatatttgg actttttcat gaccaaaaaa atgaaagcta tatggatacc 2220

ttgatccgac gtctacagct atattctcga aacctggaac atctggtaga ggaaaggaca 2280

cagctataca aggcagagag ggacagggct gacagactta actttatgtt gcttccaagg 2340

ctagtggtaa agtctctgaa ggagaaaggc tttgtagagc cggaactata tgaggaagtt 2400

acaatctact tcagtgacat tgtaggtttc actactatct gcaaatacag cacccccatg 2460

gaagtggtgg acatgcttaa tgacatctat aagagttttg accacattgt tgatcatcat 2520

gatgtctaca aggtggaaac cattggtgat gcctacatgg tggctagtgg tttgcctaag 2580

agaaatggca atcggcatgc aatagacatt gccaagatgg ccttggaaat cctcagcttc 2640

atggggacct ttgagctgga gcatcttcct ggcctcccaa tatggattcg cattggcgtt 2700

cactctggtc cctgcgctgc tggagttgtg ggaatcaaga tgcctcgtta ttgtctattt 2760

ggagatacag tcaacacagc ctctaggatg gaatccactg gcctcccttt gaggattcat 2820

gtgagtggct ccaccatagc cattctgaag agaactgagt gccagttcct gtatgaagtg 2880

agaggagaaa cgtacttaaa gggaagagga aatgagacta cctactggct gaccgggatg 2940

aaggaccaga aattcaacct gccaacccct cctactgtgg agaatcaaca gcgtttgcaa 3000

gcagaatttt cagacatgat tgccaactct ttacagaaaa gacaggcggc agggataaga 3060

agccaaaaac cgacgagt agccagctat aaaaaaggca ctctggaata cttgcaactg 3120

aataccacgg acaaggag cacctatttt acgcgtactc ggccgctcga gcagaaactc 3180

atctcagaag aggatctggc agcaaatgat atcctggatt acaaggatga cgacgataag 3240

<210> 228

<211> 1083

<212> PROTEIN

<213> Mus musculus

<400> 228

Val Phe Trp Ala Ser Gln Val Arg Gln Asn Cys Arg Asn Gly Ser Tyr

1 5 10 15

Glu Ile Ser Val Leu Met Met Asp Asn Ser Ala Tyr Lys Glu Pro Met

20 25 30

Gln Asn Leu Arg Glu Ala Val Glu Glu Gly Leu Asp Ile Val Arg Lys

35 40 45

Arg Leu Arg Glu Ala Asp Leu Asn Val Thr Val Asn Ala Thr Phe Ile

50 55 60

Tyr Ser Asp Gly Leu Ile His Lys Ser Gly Asp Cys Arg Ser Ser Thr

65 70 75 80

Cys Glu Gly Leu Asp Leu Leu Arg Glu Ile Thr Arg Asp His Lys Met

85 90 95

Gly Cys Ala Leu Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met

100 105 110

Tyr Leu Asp Thr Glu Leu Asn Tyr Pro Met Ile Ser Ala Gly Ser Tyr

115 120 125

Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Ile Leu Pro Pro

130 135 140

Ala Arg Lys Leu Met Tyr Phe Leu Val Asp Phe Trp Lys Val Asn Asn

145 150 155 160

Ala Ser Phe Lys Pro Phe Ser Trp Asn Ser Ser Tyr Val Tyr Lys Asn

165 170 175

Gly Ser Glu Pro Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala

180 185 190

Gly Val Ser Tyr Phe Ser Glu Val Leu Asn Phe Lys Asp Val Leu Arg

195 200 205

Arg Ser Glu Gln Phe Gln Glu Ile Leu Thr Gly His Asn Arg Lys Ser

210 215 220

Asn Val Ile Val Met Cys Gly Thr Pro Glu Ser Phe Tyr Asp Val Lys

225 230 235 240

Gly Asp Leu Gln Val Ala Glu Asp Thr Val Val Ile Leu Val Asp Leu

245 250 255

Phe Ser Asn His Tyr Phe Glu Glu Asn Thr Thr Ala Pro Glu Tyr Met

260 265 270

Asp Asn Val Leu Val Leu Thr Leu Pro Ser Glu Gln Ser Thr Ser Asn

275 280 285

Thr Ser Val Ala Glu Arg Phe Ser Ser Gly Arg Ser Asp Phe Ser Leu

290 295 300

Ala Tyr Leu Glu Gly Thr Leu Leu Phe Gly His Met Leu Gln Thr Phe

305 310 315 320

Leu Glu Asn Gly Glu Asn Val Thr Gly Pro Lys Phe Ala Arg Ala Phe

325 330 335

Arg Asn Leu Thr Phe Gln Gly Phe Ala Gly Pro Val Thr Leu Asp Asp

340 345 350

Ser Gly Asp Ile Asp Asn Ile Met Ser Leu Leu Tyr Val Ser Leu Asp

355 360 365

Thr Arg Lys Tyr Lys Val Leu Met Lys Tyr Asp Thr His Lys Asn Lys

370 375 380

Thr Ile Pro Val Ala Glu Asn Pro Asn Phe Ile Trp Lys Asn His Lys

385 390 395 400

Leu Pro Asn Asp Val Pro Gly Leu Gly Pro Gln Ile Leu Met Ile Ala

405 410 415

Val Phe Thr Leu Thr Gly Ile Leu Val Val Leu Leu Leu Ile Ala Leu

420 425 430

Leu Val Leu Arg Lys Tyr Arg Arg Asp His Ala Leu Arg Gln Lys Lys

435 440 445

Trp Ser His Ile Pro Ser Glu Asn Ile Phe Pro Leu Glu Thr Asn Glu

450 455 460

Thr Asn His Ile Ser Leu Lys Ile Asp Asp Asp Arg Arg Arg Asp Thr

465 470 475 480

Ile Gln Arg Val Arg Gln Cys Lys Tyr Asp Lys Lys Lys Val Ile Leu

485 490 495

Lys Asp Leu Lys His Ser Asp Gly Asn Phe Ser Glu Lys Gln Lys Ile

500 505 510

Asp Leu Asn Lys Leu Leu Gln Ser Asp Tyr Tyr Asn Leu Thr Lys Phe

515 520 525

Tyr Gly Thr Val Lys Leu Asp Thr Arg Ile Phe Gly Val Val Glu Tyr

530 535 540

Cys Glu Arg Gly Ser Leu Arg Glu Val Leu Asn Asp Thr Ile Ser Tyr

545 550 555 560

Pro Asp Gly Thr Phe Met Asp Trp Glu Phe Lys Ile Ser Val Leu Asn

565 570 575

Asp Ile Ala Lys Gly Met Ser Tyr Leu His Ser Ser Lys Ile Glu Val

580 585 590

His Gly Arg Leu Lys Ser Thr Asn Cys Val Val Asp Ser Arg Met Val

595 600 605

Val Lys Ile Thr Asp Phe Gly Cys Asn Ser Ile Leu Pro Pro Lys Lys

610 615 620

Asp Leu Trp Thr Ala Pro Glu His Leu Arg Gln Ala Thr Ile Ser Gln

625 630 635 640

Lys Gly Asp Val Tyr Ser Phe Ala Ile Ile Ala Gln Glu Ile Ile Leu

645 650 655

Arg Lys Glu Thr Phe Tyr Thr Leu Ser Cys Arg Asp His Asn Glu Lys

660 665 670

Ile Phe Arg Val Glu Asn Ser Tyr Gly Lys Pro Phe Arg Pro Asp Leu

675 680 685

Phe Leu Glu Thr Ala Asp Glu Lys Glu Leu Glu Val Tyr Leu Leu Val

690 695 700

Lys Ser Cys Trp Glu Glu Asp Pro Glu Lys Arg Pro Asp Phe Lys Lys

705 710 715 720

Ile Glu Ser Thr Leu Ala Lys Ile Phe Gly Leu Phe His Asp Gln Lys

725 730 735

Asn Glu Ser Tyr Met Asp Thr Leu Ile Arg Arg Leu Gln Leu Tyr Ser

740 745 750

Arg Asn Leu Glu His Leu Val Glu Glu Arg Thr Gln Leu Tyr Lys Ala

755 760 765

Glu Arg Asp Arg Ala Asp His Leu Asn Phe Met Leu Leu Pro Arg Leu

770 775 780

Val Val Lys Ser Leu Lys Glu Lys Gly Ile Val Glu Pro Glu Leu Tyr

785 790 795 800

Glu Glu Val Thr Ile Tyr Phe Ser Asp Ile Val Gly Phe Thr Thr Ile

805 810 815

Cys Lys Tyr Ser Thr Pro Met Glu Val Val Asp Met Leu Asn Asp Ile

820 825 830

Tyr Lys Ser Phe Asp Gln Ile Val Asp His His Asp Val Tyr Lys Val

835 840 845

Glu Thr Ile Gly Asp Ala Tyr Val Val Ala Ser Gly Leu Pro Met Arg

850 855 860

Asn Gly Asn Arg His Ala Val Asp Ile Ser Lys Met Ala Leu Asp Ile

865 870 875 880

Leu Ser Phe Ile Gly Thr Phe Glu Leu Glu His Leu Pro Gly Leu Pro

885 890 895

Val Trp Ile Arg Ile Gly Val His Ser Gly Pro Cys Ala Ala Gly Val

900 905 910

Val Gly Ile Lys Met Pro Arg Tyr Cys Leu Phe Gly Asp Thr Val Asn

915 920 925

Thr Ala Ser Arg Met Glu Ser Thr Gly Leu Pro Leu Arg Ile His Met

930 935 940

Ser Ser Ser Thr Ile Thr Ile Leu Lys Arg Thr Asp Cys Gln Phe Leu

945 950 955 960

Tyr Glu Val Arg Gly Glu Thr Tyr Leu Lys Gly Arg Gly Thr Glu Thr

965 970 975

Thr Tyr Trp Leu Thr Gly Met Lys Asp Gln Glu Tyr Asn Leu Pro Ser

980 985 990

Pro Pro Thr Val Glu Asn Gln Gln Arg Leu Gln Thr Glu Phe Ser Asp

995 1000 1005

Met Ile Val Ser Ala Leu Gln Lys Arg Gln Ala Ser Gly Lys Lys

1010 1015 1020

Ser Arg Arg Pro Thr Arg Val Ala Ser Tyr Lys Lys Gly Phe Leu

1025 1030 1035

Glu Tyr Met Gln Leu Asn Asn Ser Asp His Asp Ser Thr Tyr Phe

1040 1045 1050

Thr Arg Thr Arg Pro Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp

1055 1060 1065

Leu Ala Ala Asn Asp Ile Leu Asp Tyr Lys Asp Asp Asp Asp Lys

1070 1075 1080

<210> 229

<211> 3249

<212> DNA

<213> Mus musculus

<400> 229

gtgttctggg cctctcaggt gaggcagaac tgccgcaatg gcagctacga gatcagcgtc 60

ctgatgatgg acaactcagc ctacaaagaa cctatgcaaa acctgaggga ggctgtggag 120

gaaggactgg acatagtgcg aaagcgcctg cgtgaagccg acctaaatgt gactgtgaac 180

gcgactttca tctactccga cggtctgatt cataagtcag gtgactgccg gagcagcacc 240

tgtgaaggcc ttgacctact cagggagatt acaagagatc ataagatggg ctgcgccctc 300

atggggccct cgtgcacgta ttccaccttc cagatgtacc tcgacacaga gttgaactat 360

cccatgattt ccgctggaag ttatggattg tcctgtgact ataaggaaac cctaaccagg 420

atcctgcctc cagccaggaa gctgatgtac ttcttggtcg atttctggaa agtcaacaat 480

gcatctttca aacccttttc ctggaactct tcgtatgttt acaagaatgg atcggaacct 540

gaagattgtt tctggtacct caatgctctg gaggctgggg tgtcctattt ttctgaggtg 600

ctcaacttca aggatgtact gagacgcagc gaacagttcc aggaaatctt aacaggccat 660

aacagaaaga gcaatgtgat tgttatgtgt ggcacgccag aaagcttcta tgatgtgaaa 720

ggtgacctcc aagtggctga agatactgtt gtcatcctgg tagatctgtt cagtaaccat 780

tactttgagg agaacaccac agctcctgag tatatggaca atgtcctcgt cctgacgctg 840

ccgtctgaac agtccacctc aaacacctct gtcgccgaga ggttttcatc ggggagaagt 900

gacttttctc tcgcttactt ggagggaacc ttgctatttg gacacatgct gcagacgttt 960

cttgaaaatg gagaaaatgt cacgggtccc aagtttgctc gtgcattcag gaatctcact 1020

tttcaaggct ttgcaggacc tgtgactctg gatgacagtg gggacattga caacattatg 1080

tcccttctgt atgtgtctct ggataccagg aaatacaagg ttcttatgaa gtatgacacc 1140

cacaaaaaca aaactattcc ggtggctgag aaccccaact tcatctggaa gaaccacaag 1200

ctccccaatg acgttcctgg gctgggccct caaatcctga tgattgccgt cttcacgctc 1260

acggggatcc tggtagttct gctgctgatt gccctcctcg tgctgagaaa atacagaaga 1320

gatcatgcac ttcgacagaa gaaatggtcc cacattcctt ctgaaaacat ctttcctctg 1380

gagaccaacg agaccaacca catcagcctg aagattgacg atgacaggag acgagacaca 1440

atccagagag tgcgacagtg caaatacgac aagaagaaag tgattctgaa agacctcaag 1500

cacagcgacg ggaacttcag tgagaagcag aagatagacc tgaacaagtt gctgcagtct 1560

gactactaca acctgactaa gttctacggc accgtgaagc tggacaccag gatctttggg 1620

gtggttgagt actgcgagag gggatccctc cgggaagtgt taaacgacac aatttcctac 1680

cctgacggca cgttcatgga ttgggagttt aagatctctg tcttaaatga catcgctaag 1740

gggatgtcct acctgcactc cagtaagatt gaagtccacg ggcgtctcaa atccaccaac 1800

tgcgtggtgg acagccgcat ggtggtgaag atcaccgact ttgggtgcaa ttccatcctg 1860

cctccaaaaa aagacctgtg gacggccccg gagcacctgc gccaggccac catctctcag 1920

aaaggagacg tgtacagctt cgccatcatt gcccaggaga tcatcctccg taaggagact 1980

ttttacacgc tgagctgtcg ggatcacaat gagaagattt tcagagtgga aaattcatac 2040

gggaaacctt tccgcccaga cctcttcctg gagactgcag atgagaagga gctggaggtc 2100

tatctacttg tcaaaagctg ttgggaggag gatccagaaa agaggccaga tttcaagaaa 2160

atcgagagca cactggccaa gatatttggc cttttccatg accagaaaaa cgagtcttac 2220

atggacacct tgatccgacg tctccagctg tactctcgaa acctggaaca tctggtggag 2280

gaaaggactc agctgtacaa ggcggagagg gacagggctg accaccttaa cttcatgctc 2340

ctcccacggc tggtggtaaa gtcactgaag gagaaaggca tcgtggagcc agagctgtac 2400

gaagaagtca caatctactt cagtgacatt gtgggcttca cccacatctg caagtatagc 2460

acgcccatgg aggtggtgga catgctcaac gacatctaca agagctttga ccagattgtg 2520

gaccaccatg acgtctacaa ggtagaaacc atcggtgacg cctacgtggt ggccagcggt 2580

ctgcctatga gaaacggcaa ccgacacgcg gtagacattt ccaagatggc cttggacatc 2640

ctcagcttca tagggacctt tgagttggag catctccctg gcctccccgt gtggatccgc 2700

attggagttc attctgggcc ctgcgctgct ggtgttgtgg ggatcaagat gcctcgctat 2760

tgcctgtttg gagacactgt caacactgcc tccaggatgg aatccaccgg cctccccttg 2820

aggattcaca tgagcagctc caccataacc atcctgaaga gaacggattg ccagttcctg 2880

tatgaagtga ggggagaaac ctacttaaag ggaagaggga cagagaccac atactggctg 2940

actgggatga aggaccaaga atacaacctg ccatccccac cgacagtgga gaaccaacag 3000

cgtctgcaga ctgagttctc agacatgatc gttagcgcct tacagaaaag acaggcctcg 3060

ggcaagaaga gccggaggcc cactcgggtg gccagctaca agaaaggctt tctggaatac 3120

atgcagctga acaattcaga ccacgatagc acctatttta cgcgtacgcg gccgctcgag 3180

cagaaactca tctcagaaga ggatctggca gcaaatgata tcctggatta caaggatgac 3240

gacgataag 3249

<210> 230

<211> 672

<212> PROTEIN

<213> Homo sapiens

<400> 230

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val Leu

85 90 95

Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Thr

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Ile

210 215 220

Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Gly Arg Gly Pro Gln Gly Gly Gly Gly Ser Glu Asn Leu Tyr

405 410 415

Phe Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Arg Gly

420 425 430

Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro Asn Leu

435 440 445

Glu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp Val

450 455 460

Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val Val Asp Val

465 470 475 480

Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val

485 490 495

Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser

500 505 510

Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp Met

515 520 525

Ser Gly Lys Ala Phe Ala Cys Ala Val Asn Asn Lys Asp Leu Pro Ala

530 535 540

Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg Ala Pro

545 550 555 560

Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Lys Gln

565 570 575

Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp Ile Tyr

580 585 590

Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys Asn Thr

595 600 605

Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser Lys Leu

610 615 620

Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser Cys Ser

625 630 635 640

Val Val His Glu Gly Leu His Asn His His Thr Thr Lys Ser Phe Ser

645 650 655

Arg Thr Pro Gly Gly Gly Pro Pro Asp Tyr Lys Asp Asp Asp Asp Lys

660 665 670

<210> 231

<211> 2076

<212> DNA

<213> Homo sapiens

<400> 231

atggagacag acacactgct cctgtgggtg ctgcttctgt gggtgccagg ttccactgga 60

tcacaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatgggc 120

aactcagcct ttgcagagcc cctgaaaaac ttggaagatg cggtgaatga ggggctggaa 180

atagtgagag gacgtctgca aaatgctggc ctaaatgtga ctgtgaacgc tactttcatg 240

tattcggatg gtctgattca taactcaggc gactgccgga gtagcacctg tgaaggcctc 300

gacctactca ggaaaatttc aaatgcacaa cggatgggct gtgtcctcat agggccctca 360

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 420

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 480

gctagaaagt tgatgtactt cttggttaac ttttggaaaa ccaacgatct gcccttcaaa 540

acttattcct ggagcacttc gtatgtttac aagaatggta cagaaactga ggactgtttc 600

tggtacctta atgctctgga ggctagcgtt tcctatttct cccacgaact cggctttaag 660

gtggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 720

aatgtgatta ttatgtgtgg tggtccagag ttcctctaca agctgaaggg tgaccgagca 780

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta ctttgaggac 840

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgctgtc tcctgggaat 900

tcccttctaa atagctcttt ctccaggaat ctatcaccaa caaaacgaga ctttgctctt 960

gcctatttga atggaatcct gctctttgga catatgctga agatatttct tgaaaatgga 1020

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1080

gacggtccag tgaccttgga tgactggggg gatgttgaca gtaccatggt gcttctgtat 1140

acctctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaataag 1200

acctatcctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1260

attacaggcc ggggccctca gggaggcgga ggttccgaga atttatactt ccagggcgga 1320

ggcggttccg gcggcggagg aagcgagccc cgcggaccga caatcaagcc ctgtcctcca 1380

tgcaaatgcc cagcacctaa cctcgagggt ggaccatccg tcttcatctt ccctccaaag 1440

atcaaggatg tactcatgat ctccctgagc cccatagtca catgtgtggt ggtggatgtg 1500

agcgaggatg acccagatgt ccagatcagc tggtttgtga acaacgtgga agtacacaca 1560

gctcagacac aaacccatag agaggattac aacagtactc tccgggtggt cagtgccctc 1620

cccatccagc accaggactg gatgagtggc aaggctttcg catgcgccgt caacaacaaa 1680

gacctcccag cgcccatcga gagaaccatc tcaaaaccca aagggtcagt aagagctcca 1740

caggtatatg tcttgcctcc accagaagaa gagatgacta agaaacaggt cactctgacc 1800

tgcatggtca cagacttcat gcctgaagac atttacgtgg agtggaccaa caacgggaaa 1860

acagagctaa actacaagaa cactgaacca gtcctggact ctgatggttc ttacttcatg 1920

tacagcaagc tgagagtgga aaagaagaac tgggtggaaa gaaatagcta ctcctgttca 1980

gtggtccacg agggtctgca caatcaccac acgactaaga gcttctcccg gactccgggt 2040

ggcgggccac ccgactacaa ggacgacgat gacaaa 2076

<210> 232

<211> 418

<212> PROTEIN

<213> Homo sapiens

<400> 232

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val Leu

85 90 95

Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Thr

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Ile

210 215 220

Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Gly Arg Gly Pro Gln Gly Gly Gly Gly Ser Glu Asn Leu Tyr

405 410 415

Phe Gln

<210> 233

<211> 1254

<212> DNA

<213> Homo sapiens

<400> 233

tcacaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatgggc 60

aactcagcct ttgcagagcc cctgaaaaac ttggaagatg cggtgaatga ggggctggaa 120

atagtgagag gacgtctgca aaatgctggc ctaaatgtga ctgtgaacgc tactttcatg 180

tattcggatg gtctgattca taactcaggc gactgccgga gtagcacctg tgaaggcctc 240

gacctactca ggaaaatttc aaatgcacaa cggatgggct gtgtcctcat agggccctca 300

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 360

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 420

gctagaaagt tgatgtactt cttggttaac ttttggaaaa ccaacgatct gcccttcaaa 480

acttattcct ggagcacttc gtatgtttac aagaatggta cagaaactga ggactgtttc 540

tggtacctta atgctctgga ggctagcgtt tcctatttct cccacgaact cggctttaag 600

gtggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 660

aatgtgatta ttatgtgtgg tggtccagag ttcctctaca agctgaaggg tgaccgagca 720

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta ctttgaggac 780

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgctgtc tcctgggaat 840

tcccttctaa atagctcttt ctccaggaat ctatcaccaa caaaacgaga ctttgctctt 900

gcctatttga atggaatcct gctctttgga catatgctga agatatttct tgaaaatgga 960

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1020

gacggtccag tgaccttgga tgactgggg gatgttgaca gtaccatggt gcttctgtat 1080

acctctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaataag 1140

acctatcctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1200

attacaggcc ggggccctca gggaggcgga ggttccgaga atttatactt ccag 1254

<210> 234

<211> 672

<212> PROTEIN

<213> Macaca fascicularis

<400> 234

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Asp Asn Ser Ala Phe Ala Glu Pro Leu Glu Asn Val Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Ser Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Lys Arg Met Gly Cys Val Leu

85 90 95

Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Thr Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Ser

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Ser Phe Lys Leu Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Val

210 215 220

Met Cys Gly Asp Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Gln Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Thr Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Gln Thr Asn Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Asp Arg Gly Pro Gln Gly Gly Gly Gly Ser Glu Asn Leu Tyr

405 410 415

Phe Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Pro Arg Gly

420 425 430

Pro Thr Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro Asn Leu

435 440 445

Glu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp Val

450 455 460

Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val Val Asp Val

465 470 475 480

Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe Val Asn Asn Val

485 490 495

Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn Ser

500 505 510

Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp Met

515 520 525

Ser Gly Lys Ala Phe Ala Cys Ala Val Asn Asn Lys Asp Leu Pro Ala

530 535 540

Pro Ile Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg Ala Pro

545 550 555 560

Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Lys Gln

565 570 575

Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp Ile Tyr

580 585 590

Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr Lys Asn Thr

595 600 605

Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser Lys Leu

610 615 620

Arg Val Glu Lys Lys Asn Trp Val Glu Arg Asn Ser Tyr Ser Cys Ser

625 630 635 640

Val Val His Glu Gly Leu His Asn His His Thr Thr Lys Ser Phe Ser

645 650 655

Arg Thr Pro Gly Gly Gly Pro Pro Asp Tyr Lys Asp Asp Asp Asp Lys

660 665 670

<210> 235

<211> 2016

<212> DNA

<213> Macaca fascicularis

<400> 235

tcacaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatggac 60

aactcagcct ttgcagagcc cctggaaaac gtggaagatg cggtgaatga ggggctggaa 120

atagtgagag gacgtctgca aaacgctggc ctaaatgtga ctgtgaatgc ttctttcatg 180

tattcggatg gtctgattca taactccggc gactgccgga gcagcacctg tgaaggcctt 240

gacctactca ggaaaatttc aaatgcaaaa cggatgggct gtgtcctcat ggggccctca 300

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 360

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 420

gctagaaagt tgacatactt cttggttaac ttttggaaaa ccaatgatct acccttcaaa 480

acttattcct ggagcacttc gtatgtttac aagaatggta cggagtccga ggactgtttc 540

tggtacctta acgctctgga ggccagtgtt tcctatttct cccacgaact cagttttaag 600

ttggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 660

aatgtgattg ttatgtgtgg tgatccagag ttcctctaca agttgaaggg tgaccgagca 720

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta ctttgaggac 780

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgcagtc tcctgggaat 840

tccctcctaa atagctcttt ctccaggaat ctatccccaa caaaacgaga ctttgctctt 900

gcctatttga atggaatcct gctctttgga catatgctaa agacatttct tgaaaatgga 960

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1020

gacggtccag tgaccttgga tgactgggg gatgtggaca gtaccatggt gcttctgtat 1080

acgtctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaatcag 1140

accaaccctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1200

attacagacc ggggccctca gggaggcgga ggttccgaga atttatactt ccagggcgga 1260

ggcggttccg gcggcggagg aagcgagccc cgcggaccga caatcaagcc ctgtcctcca 1320

tgcaaatgcc cagcacctaa cctcgagggt ggaccatccg tcttcatctt ccctccaaag 1380

atcaaggatg tactcatgat ctccctgagc cccatagtca catgtgtggt ggtggatgtg 1440

agcgaggatg acccagatgt ccagatcagc tggtttgtga acaacgtgga agtacacaca 1500

gctcagacac aaacccatag agaggattac aacagtactc tccgggtggt cagtgccctc 1560

cccatccagc accaggactg gatgagtggc aaggctttcg catgcgccgt caacaacaaa 1620

gacctcccag cgcccatcga gagaaccatc tcaaaaccca aagggtcagt aagagctcca 1680

caggtatatg tcttgcctcc accagaagaa gagatgacta agaaacaggt cactctgacc 1740

tgcatggtca cagacttcat gcctgaagac atttacgtgg agtggaccaa caacgggaaa 1800

acagagctaa actacaagaa cactgaacca gtcctggact ctgatggttc ttacttcatg 1860

tacagcaagc tgagagtgga aaagaagaac tgggtggaaa gaaatagcta ctcctgttca 1920

gtggtccacg agggtctgca caatcaccac acgactaaga gcttctcccg gactccgggt 1980

ggcgggccac ccgactacaa ggacgacgat gacaaa 2016

<210> 236

<211> 418

<212> PROTEIN

<213> Macaca fascicularis

<400> 236

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Asp Asn Ser Ala Phe Ala Glu Pro Leu Glu Asn Val Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Ser Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Lys Arg Met Gly Cys Val Leu

85 90 95

Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Thr Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Ser

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Ser Phe Lys Leu Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Val

210 215 220

Met Cys Gly Asp Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Gln Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Thr Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Gln Thr Asn Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Asp Arg Gly Pro Gln Gly Gly Gly Gly Ser Glu Asn Leu Tyr

405 410 415

Phe Gln

<210> 237

<211> 1254

<212> DNA

<213> Macaca fascicularis

<400> 237

tcacaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatggac 60

aactcagcct ttgcagagcc cctggaaaac gtggaagatg cggtgaatga ggggctggaa 120

atagtgagag gacgtctgca aaacgctggc ctaaatgtga ctgtgaatgc ttctttcatg 180

tattcggatg gtctgattca taactccggc gactgccgga gcagcacctg tgaaggcctt 240

gacctactca ggaaaatttc aaatgcaaaa cggatgggct gtgtcctcat ggggccctca 300

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 360

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 420

gctagaaagt tgacatactt cttggttaac ttttggaaaa ccaatgatct acccttcaaa 480

acttattcct ggagcacttc gtatgtttac aagaatggta cggagtccga ggactgtttc 540

tggtacctta acgctctgga ggccagtgtt tcctatttct cccacgaact cagttttaag 600

ttggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 660

aatgtgattg ttatgtgtgg tgatccagag ttcctctaca agttgaaggg tgaccgagca 720

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta ctttgaggac 780

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgcagtc tcctgggaat 840

tccctcctaa atagctcttt ctccaggaat ctatccccaa caaaacgaga ctttgctctt 900

gcctatttga atggaatcct gctctttgga catatgctaa agacatttct tgaaaatgga 960

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1020

gacggtccag tgaccttgga tgactgggg gatgtggaca gtaccatggt gcttctgtat 1080

acgtctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaatcag 1140

accaaccctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1200

attacagacc ggggccctca gggaggcgga ggttccgaga atttatactt ccag 1254

<210> 238

<211> 673

<212> PROTEIN

<213> Mus musculus

<400> 238

Val Phe Trp Ala Ser Gln Val Arg Gln Asn Cys Arg Asn Gly Ser Tyr

1 5 10 15

Glu Ile Ser Val Leu Met Met Asp Asn Ser Ala Tyr Lys Glu Pro Met

20 25 30

Gln Asn Leu Arg Glu Ala Val Glu Glu Gly Leu Asp Ile Val Arg Lys

35 40 45

Arg Leu Arg Glu Ala Asp Leu Asn Val Thr Val Asn Ala Thr Phe Ile

50 55 60

Tyr Ser Asp Gly Leu Ile His Lys Ser Gly Asp Cys Arg Ser Ser Thr

65 70 75 80

Cys Glu Gly Leu Asp Leu Leu Arg Glu Ile Thr Arg Asp His Lys Met

85 90 95

Gly Cys Ala Leu Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met

100 105 110

Tyr Leu Asp Thr Glu Leu Asn Tyr Pro Met Ile Ser Ala Gly Ser Tyr

115 120 125

Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Ile Leu Pro Pro

130 135 140

Ala Arg Lys Leu Met Tyr Phe Leu Val Asp Phe Trp Lys Val Asn Asn

145 150 155 160

Ala Ser Phe Lys Pro Phe Ser Trp Asn Ser Ser Tyr Val Tyr Lys Asn

165 170 175

Gly Ser Glu Pro Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala

180 185 190

Gly Val Ser Tyr Phe Ser Glu Val Leu Asn Phe Lys Asp Val Leu Arg

195 200 205

Arg Ser Glu Gln Phe Gln Glu Ile Leu Thr Gly His Asn Arg Lys Ser

210 215 220

Asn Val Ile Val Met Cys Gly Thr Pro Glu Ser Phe Tyr Asp Val Lys

225 230 235 240

Gly Asp Leu Gln Val Ala Glu Asp Thr Val Val Ile Leu Val Asp Leu

245 250 255

Phe Ser Asn His Tyr Phe Glu Glu Asn Thr Thr Ala Pro Glu Tyr Met

260 265 270

Asp Asn Val Leu Val Leu Thr Leu Pro Ser Glu Gln Ser Thr Ser Asn

275 280 285

Thr Ser Val Ala Glu Arg Phe Ser Ser Gly Arg Ser Asp Phe Ser Leu

290 295 300

Ala Tyr Leu Glu Gly Thr Leu Leu Phe Gly His Met Leu Gln Thr Phe

305 310 315 320

Leu Glu Asn Gly Glu Asn Val Thr Gly Pro Lys Phe Ala Arg Ala Phe

325 330 335

Arg Asn Leu Thr Phe Gln Gly Phe Ala Gly Pro Val Thr Leu Asp Asp

340 345 350

Ser Gly Asp Ile Asp Asn Ile Met Ser Leu Leu Tyr Val Ser Leu Asp

355 360 365

Thr Arg Lys Tyr Lys Val Leu Met Lys Tyr Asp Thr His Lys Asn Lys

370 375 380

Thr Ile Pro Val Ala Glu Asn Pro Asn Phe Ile Trp Lys Asn His Lys

385 390 395 400

Leu Pro Asn Asp Val Pro Gly Leu Gly Pro Gln Ile Leu Met Gly Gly

405 410 415

Gly Gly Ser Glu Asn Leu Tyr Phe Gln Gly Gly Gly Gly Ser Gly Gly

420 425 430

Gly Gly Ser Glu Pro Arg Gly Pro Thr Ile Lys Pro Cys Pro Pro Cys

435 440 445

Lys Cys Pro Ala Pro Asn Leu Glu Gly Gly Pro Ser Val Phe Ile Phe

450 455 460

Pro Pro Lys Ile Lys Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val

465 470 475 480

Thr Cys Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile

485 490 495

Ser Trp Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr

500 505 510

His Arg Glu Asp Tyr Asn Ser Thr Leu Arg Val Val Ser Ala Leu Pro

515 520 525

Ile Gln His Gln Asp Trp Met Ser Gly Lys Ala Phe Ala Cys Ala Val

530 535 540

Asn Asn Lys Asp Leu Pro Ala Pro Ile Glu Arg Thr Ile Ser Lys Pro

545 550 555 560

Lys Gly Ser Val Arg Ala Pro Gln Val Tyr Val Leu Pro Pro Pro Glu

565 570 575

Glu Glu Met Thr Lys Lys Gln Val Thr Leu Thr Cys Met Val Thr Asp

580 585 590

Phe Met Pro Glu Asp Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr

595 600 605

Glu Leu Asn Tyr Lys Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser

610 615 620

Tyr Phe Met Tyr Ser Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu

625 630 635 640

Arg Asn Ser Tyr Ser Cys Ser Val Val His Glu Gly Leu His Asn His

645 650 655

His Thr Thr Lys Ser Phe Ser Arg Thr Pro Gly His His His His

660 665 670

His

<210> 239

<211> 2019

<212> DNA

<213> Mus musculus

<400> 239

gtgttctggg cctctcaggt gaggcagaac tgccgcaatg gcagctacga gatcagcgtc 60

ctgatgatgg acaactcagc ctacaaagaa cctatgcaaa acctgaggga ggctgtggag 120

gaaggactgg acatagtgcg aaagcgcctg cgtgaagccg acctaaatgt gactgtgaac 180

gcgactttca tctactccga cggtctgatt cataagtcag gtgactgccg gagcagcacc 240

tgtgaaggcc ttgacctact cagggagatt acaagagatc ataagatggg ctgcgccctc 300

atggggccct cgtgcacgta ttccaccttc cagatgtacc tcgacacaga gttgaactat 360

cccatgattt ccgctggaag ttatggattg tcctgtgact ataaggaaac cctaaccagg 420

attctgcctc cagccaggaa gctgatgtac ttcttggtcg atttctggaa agtcaacaat 480

gcatctttca aacccttttc ctggaactct tcgtatgttt acaagaatgg atcggaacct 540

gaagattgtt tctggtacct caatgctctg gaggctgggg tgtcctattt ttctgaggtg 600

ctcaacttca aggatgtact gagacgcagc gaacagttcc aggaaatctt aacaggccat 660

aacagaaaga gcaatgtgat tgttatgtgt ggcacgccag aaagcttcta tgatgtgaaa 720

ggtgacctcc aagtggctga agatactgtt gtcatcctgg tagatctgtt cagtaaccat 780

tactttgagg agaacaccac agctcctgag tatatggaca atgtcctcgt cctgacgctg 840

ccgtctgaac agtccacctc aaacacctct gtcgccgaga ggttttcatc ggggagaagt 900

gacttttctc tcgcttactt ggagggaacc ttgctatttg gacacatgct gcagacgttt 960

cttgaaaatg gagaaaatgt cacgggtccc aagtttgctc gtgcattcag gaatctcact 1020

tttcaaggct ttgcaggacc tgtgactctg gatgacagtg gggacattga caacattatg 1080

tcccttctgt atgtgtctct ggataccagg aaatacaagg ttcttatgaa gtatgacacc 1140

cacaaaaaca aaactattcc ggtggctgag aaccccaact tcatctggaa gaaccacaag 1200

ctccccaatg acgttcctgg gctgggccct caaatcctga tgggaggcgg aggttccgag 1260

aatttatact tccagggcgg aggcggttcc ggcggcggag gaagcgagcc ccgcggaccg 1320

acaatcaagc cctgtcctcc atgcaaatgc ccagcaccta acctcgaggg tggaccatcc 1380

gtcttcatct tccctccaaa gatcaaggat gtactcatga tctccctgag ccccatagtc 1440

acatgtgtgg tggtggatgt gagcgaggat gacccagatg tccagatcag ctggtttgtg 1500

aacaacgtgg aagtacacac agctcagaca caaacccata gagaggatta caacagtact 1560

ctccgggtgg tcagtgccct ccccatccag caccaggact ggatgagtgg caaggctttc 1620

gcatgcgccg tcaacaacaa agacctccca gcgcccatcg agagaaccat ctcaaaaccc 1680

aaagggtcag taagagctcc acaggtatat gtcttgcctc caccagaaga agagatgact 1740

aagaaacagg tcactctgac ctgcatggtc acagacttca tgcctgaaga catttacgtg 1800

gagtggacca acaacgggaa aacagagcta aactacaaga acactgaacc agtcctggac 1860

tctgatggtt cttacttcat gtacagcaag ctgagagtgg aaaagaagaa ctgggtggaa 1920

agaaatagct actcctgttc agtggtccac gagggtctgc acaatcacca cacgactaag 1980

agcttctccc ggactccggg tcaccatcac catcaccat 2019

<210> 240

<211> 425

<212> PROTEIN

<213> Mus musculus

<400> 240

Val Phe Trp Ala Ser Gln Val Arg Gln Asn Cys Arg Asn Gly Ser Tyr

1 5 10 15

Glu Ile Ser Val Leu Met Met Asp Asn Ser Ala Tyr Lys Glu Pro Met

20 25 30

Gln Asn Leu Arg Glu Ala Val Glu Glu Gly Leu Asp Ile Val Arg Lys

35 40 45

Arg Leu Arg Glu Ala Asp Leu Asn Val Thr Val Asn Ala Thr Phe Ile

50 55 60

Tyr Ser Asp Gly Leu Ile His Lys Ser Gly Asp Cys Arg Ser Ser Thr

65 70 75 80

Cys Glu Gly Leu Asp Leu Leu Arg Glu Ile Thr Arg Asp His Lys Met

85 90 95

Gly Cys Ala Leu Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met

100 105 110

Tyr Leu Asp Thr Glu Leu Asn Tyr Pro Met Ile Ser Ala Gly Ser Tyr

115 120 125

Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Ile Leu Pro Pro

130 135 140

Ala Arg Lys Leu Met Tyr Phe Leu Val Asp Phe Trp Lys Val Asn Asn

145 150 155 160

Ala Ser Phe Lys Pro Phe Ser Trp Asn Ser Ser Tyr Val Tyr Lys Asn

165 170 175

Gly Ser Glu Pro Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala

180 185 190

Gly Val Ser Tyr Phe Ser Glu Val Leu Asn Phe Lys Asp Val Leu Arg

195 200 205

Arg Ser Glu Gln Phe Gln Glu Ile Leu Thr Gly His Asn Arg Lys Ser

210 215 220

Asn Val Ile Val Met Cys Gly Thr Pro Glu Ser Phe Tyr Asp Val Lys

225 230 235 240

Gly Asp Leu Gln Val Ala Glu Asp Thr Val Val Ile Leu Val Asp Leu

245 250 255

Phe Ser Asn His Tyr Phe Glu Glu Asn Thr Thr Ala Pro Glu Tyr Met

260 265 270

Asp Asn Val Leu Val Leu Thr Leu Pro Ser Glu Gln Ser Thr Ser Asn

275 280 285

Thr Ser Val Ala Glu Arg Phe Ser Ser Gly Arg Ser Asp Phe Ser Leu

290 295 300

Ala Tyr Leu Glu Gly Thr Leu Leu Phe Gly His Met Leu Gln Thr Phe

305 310 315 320

Leu Glu Asn Gly Glu Asn Val Thr Gly Pro Lys Phe Ala Arg Ala Phe

325 330 335

Arg Asn Leu Thr Phe Gln Gly Phe Ala Gly Pro Val Thr Leu Asp Asp

340 345 350

Ser Gly Asp Ile Asp Asn Ile Met Ser Leu Leu Tyr Val Ser Leu Asp

355 360 365

Thr Arg Lys Tyr Lys Val Leu Met Lys Tyr Asp Thr His Lys Asn Lys

370 375 380

Thr Ile Pro Val Ala Glu Asn Pro Asn Phe Ile Trp Lys Asn His Lys

385 390 395 400

Leu Pro Asn Asp Val Pro Gly Leu Gly Pro Gln Ile Leu Met Gly Gly

405 410 415

Gly Gly Ser Glu Asn Leu Tyr Phe Gln

420 425

<210> 241

<211> 1275

<212> DNA

<213> Mus musculus

<400> 241

gtgttctggg cctctcaggt gaggcagaac tgccgcaatg gcagctacga gatcagcgtc 60

ctgatgatgg acaactcagc ctacaaagaa cctatgcaaa acctgaggga ggctgtggag 120

gaaggactgg acatagtgcg aaagcgcctg cgtgaagccg acctaaatgt gactgtgaac 180

gcgactttca tctactccga cggtctgatt cataagtcag gtgactgccg gagcagcacc 240

tgtgaaggcc ttgacctact cagggagatt acaagagatc ataagatggg ctgcgccctc 300

atggggccct cgtgcacgta ttccaccttc cagatgtacc tcgacacaga gttgaactat 360

cccatgattt ccgctggaag ttatggattg tcctgtgact ataaggaaac cctaaccagg 420

attctgcctc cagccaggaa gctgatgtac ttcttggtcg atttctggaa agtcaacaat 480

gcatctttca aacccttttc ctggaactct tcgtatgttt acaagaatgg atcggaacct 540

gaagattgtt tctggtacct caatgctctg gaggctgggg tgtcctattt ttctgaggtg 600

ctcaacttca aggatgtact gagacgcagc gaacagttcc aggaaatctt aacaggccat 660

aacagaaaga gcaatgtgat tgttatgtgt ggcacgccag aaagcttcta tgatgtgaaa 720

ggtgacctcc aagtggctga agatactgtt gtcatcctgg tagatctgtt cagtaaccat 780

tactttgagg agaacaccac agctcctgag tatatggaca atgtcctcgt cctgacgctg 840

ccgtctgaac agtccacctc aaacacctct gtcgccgaga ggttttcatc ggggagaagt 900

gacttttctc tcgcttactt ggagggaacc ttgctatttg gacacatgct gcagacgttt 960

cttgaaaatg gagaaaatgt cacgggtccc aagtttgctc gtgcattcag gaatctcact 1020

tttcaaggct ttgcaggacc tgtgactctg gatgacagtg gggacattga caacattatg 1080

tcccttctgt atgtgtctct ggataccagg aaatacaagg ttcttatgaa gtatgacacc 1140

cacaaaaaca aaactattcc ggtggctgag aaccccaact tcatctggaa gaaccacaag 1200

ctccccaatg acgttcctgg gctgggccct caaatcctga tgggaggcgg aggttccgag 1260

aatttactact tccag 1275

<210> 242

<211> 214

<212> PROTEIN

<213> Homo sapiens

<400> 242

Met Gln Ser Gly Thr His Trp Arg Val Leu Gly Leu Cys Leu Leu Ser

1 5 10 15

Val Gly Val Trp Gly Gln Asp Gly Asn Glu Glu Met Gly Gly Ile Thr

20 25 30

Gln Thr Pro Tyr Lys Val Ser Ile Ser Gly Thr Thr Val Ile Leu Thr

35 40 45

Cys Pro Gln Tyr Pro Gly Ser Glu Ile Leu Trp Gln His Asn Asp Lys

50 55 60

Asn Ile Gly Gly Asp Glu Asp Asp Lys Asn Ile Gly Ser Asp Glu Asp

65 70 75 80

His Leu Ser Leu Lys Glu Phe Ser Glu Leu Glu Gln Ser Gly Tyr Tyr

85 90 95

Val Cys Tyr Pro Arg Gly Ser Lys Pro Glu Asp Ala Asn Phe Tyr Leu

100 105 110

Tyr Leu Arg Ala Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Pro Ile Glu Glu Leu Glu Asp Arg Val Phe Val Asn

130 135 140

Cys Asn Thr Ser Ile Thr Trp Val Glu Gly Thr Val Gly Thr Leu Leu

145 150 155 160

Ser Asp Ile Thr Arg Leu Asp Leu Gly Lys Arg Ile Leu Asp Pro Arg

165 170 175

Gly Ile Tyr Arg Cys Asn Gly Thr Asp Ile Tyr Lys Asp Lys Glu Ser

180 185 190

Thr Val Gln Val His Tyr Arg Met Cys Gln Ser Cys Val Glu Leu Asp

195 200 205

His His His His His

210

<210> 243

<211> 642

<212> DNA

<213> Homo sapiens

<400> 243

atgcaatccg gtacgcactg gagagtcttg ggtctgtgcc ttttgtctgt tggcgtatgg 60

gggcaagacg ggaacgaaga aatgggaggc attacacaaa caccatacaa ggtatcaatt 120

agcggcacga cggttatact gacatgtcca caatatccag gcagcgaaat tctgtggcag 180

cacaatgaca agaatattgg gggagatgaa gacgacaaaa atatcggtag cgacgaggac 240

catctgtctc tgaaggaatt ttcagaactt gaacaatctg gctattatgt gtgctacccg 300

cgaggcagca aaccggaaga tgcaaacttt tacctttatc tgagagcaag gggcggcgga 360

ggctctgggg gcggaggcag cggcggagga ggatcaccaa tcgaggaatt ggaagatagg 420

gtattcgtaa attgtaacac cagcattaca tgggtggaag ggaccgttgg aactctcctg 480

agcgatatca cacgactgga tcttggtaaa cgaatcctgg acccacgcgg aatctataga 540

tgtaacggaa ctgatattta caaagacaaa gaatctactg tgcaagttca ctaccgaatg 600

tgtcaatcat gcgttgaact cgatcaccac caccatcatc ac 642

<210> 244

<211> 205

<212> PROTEIN

<213> Macaca fascicularis

<400> 244

Met Gln Ser Gly Thr His Trp Arg Val Leu Gly Leu Cys Leu Leu Ser

1 5 10 15

Val Gly Val Trp Gly Gln Asp Gly Asn Glu Glu Met Gly Ser Ile Thr

20 25 30

Gln Thr Pro Tyr Gln Val Ser Ile Ser Gly Thr Thr Val Ile Leu Thr

35 40 45

Cys Ser Gln His Leu Gly Ser Glu Ala Gln Trp Gln His Asn Gly Lys

50 55 60

Asn Lys Gly Asp Ser Gly Asp Gln Leu Phe Leu Pro Glu Phe Ser Glu

65 70 75 80

Met Glu Gln Ser Gly Tyr Tyr Val Cys Tyr Pro Arg Gly Ser Asn Pro

85 90 95

Glu Asp Ala Ser His His Leu Tyr Leu Lys Ala Arg Gly Gly Gly Gly

100 105 110

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Pro Val Glu Glu Leu

115 120 125

Glu Asp Arg Val Phe Val Lys Cys Asn Thr Ser Val Thr Trp Val Glu

130 135 140

Gly Thr Val Gly Thr Leu Leu Thr Asn Asn Thr Arg Leu Asp Leu Gly

145 150 155 160

Lys Arg Ile Leu Asp Pro Arg Gly Ile Tyr Arg Cys Asn Gly Thr Asp

165 170 175

Ile Tyr Lys Asp Lys Glu Ser Ala Val Gln Val His Tyr Arg Met Cys

180 185 190

Gln Asn Cys Val Glu Leu Asp His His His His His

195 200 205

<210> 245

<211> 615

<212> DNA

<213> Macaca fascicularis

<400> 245

atgcaaagcg gaactcattg gcgcgtcctg ggactctgtc tgctctccgt gggagtctgg 60

ggacaagatg gaaacgaaga gatgggaagc attacacaaa caccatacca agtctccatt 120

agcggcacta ccgtcattct gacatgttcc caacatctgg gcagcgaagc tcaatggcag 180

cacaatggaa agaataaggg cgatagcgga gaccaactgt ttctgccaga atttagcgaa 240

atggagcaat ccggctatta cgtgtgctac ccacgcggca gcaaccctga agatgctagc 300

catcacctct atctgaaggc tcgcggaggc ggaggcagcg gcggcggagg atccggcgga 360

ggcggaagcc cagtcgagga actggaagat cgcgtcttcg tgaagtgtaa caccagcgtc 420

acatgggtgg aaggcaccgt cggaactctc ctgactaaca acacacgcct ggatctcgga 480

aaacgcatcc tggacccacg cggaatctat agatgtaacg gaactgatat ttacaaagac 540

aaagaatccg ctgtgcaagt ccactaccgc atgtgtcaaa actgtgtcga actggatcat 600

caccatcacc atcac 615

<210> 246

<211> 1386

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 246

gacatccagc tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgca gagccagtga aagtgttgat tattatggca gtagtttatt gcagtggtat 120

cagcagaaac cagggaaagc ccctaagctc ctgatctatg ctgcatccaa actagcttct 180

ggggtcccat caaggttcag tggcagtgga tctgggacag atttcactct caccatcagc 240

agtctgcaac ctgaagattt tgcaacttac tactgtcagc aaactcggaa agcgtatacg 300

tttggccagg ggaccaagct ggagatcaaa ggtggaggta gcggggggcgg cggggaagtt 360

caactcgttg agtctggcgg gggattggtt caacccggtg gaagccttag attgtcatgt 420

gccgcctccg gctttacatt tagcgactat tacatgacct gggtgagaca agctccaggc 480

aaaggacttg aatgggtggc ctttatcaga aatcaggccc gcggctacac aagcgaccat 540

aatccctccg tgaaaggaag atttaccatc agccgggaca atgctaaaaa ttcactttac 600

cttcaaatga actctcttag agccgaggac accgccgtat actactgcgc aagagataga 660

ccaagttatt acgtcctgga ttactggggc cagggaacaa ccgtcaccgt gtcttctgga 720

tgcccaccgt gcccagcacc tgaagccgct ggggcaccgt cagtcttcct cttcccccca 780

aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac 840

gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 900

aatgccaaga caaagccgcg ggagggagcag tacaacagca cgtaccgtgt ggtcagcgtc 960

ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac 1020

aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa 1080

ccacaggtgt gcaccctgcc cccacccgg gaggagatga ccaagaacca ggtcagcctg 1140

tggtgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg 1200

cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc 1260

ctctatagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc 1320

tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtccccc 1380

ggaaag 1386

<210> 247

<211> 1398

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 247

gacatccaga tgacccagtc cccctcttct ctgtctgcct ctgtgggcga cagagtgacc 60

atcacctgca caagctcaca gtcactgttt aatgtccgca gccagaaaaa ctatcttgcg 120

tggtatcagc agaagcctgg caaggctccc aagctgctga tctactgggc cagtacacga 180

gaatccggcg tgccttccag attctccggc tctggctctg gcaccgattt caccctgacc 240

atctcctccc tccagcctga ggatttcgcc acctactact gcaaacagtc ttacgacctt 300

ttcacttttg gcggcggaac aaaggtggag atcaagggcg gaggtggatc tggcggcgga 360

ggcgaggtgc agctggtgga gtctggggga ggcttggtcc agcctggggg gtccctgaga 420

ctctcctgtg cagcctctgg cttcaccttc agcagctact ggatgcactg ggtccgccag 480

gctccaggga aggggctgga gtggattgga gagattaaac ctagcaacga acttactaac 540

gtccatgaaa agttcaagga ccgattcacc atctccgtgg acaaggccaa gaactcagcc 600

tatctgcaaa tgaacagcct gagagccgag gacacggctg tgtattactg tacaagaacg 660

attacgacga cggagggata ctggttcttc gatgtctggg gccaagggac actggtcacc 720

gtctcttcag gatgtccacc gtgcccagca cctgaagccg ctggggcacc gtcagtcttc 780

ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 840

gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 900

gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 960

gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 1020

aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1080

cagccccgag aaccacaggt gtacaccctg cccccatgcc gggaggagat gaccaagaac 1140

caggtcagcc tgtcctgcgc ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1200

gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1260

ggctccttct tcctcgttag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1320

gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1380

tccctgtccc ccggaaag 1398

<210> 248

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 248

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 249

<211> 464

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 249

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Arg Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly

115 120 125

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

130 135 140

Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala

145 150 155 160

Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly

165 170 175

Leu Thr Asn Tyr Ile Glu Lys Phe Lys Asn Arg Phe Thr Ile Ser Val

180 185 190

Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu

210 215 220

Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val

225 230 235 240

Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro

245 250 255

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

260 265 270

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

275 280 285

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

290 295 300

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

305 310 315 320

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

325 330 335

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

340 345 350

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

355 360 365

Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser

370 375 380

Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

385 390 395 400

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

405 410 415

Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys

420 425 430

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

435 440 445

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 250

<211> 925

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 250

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Arg Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Asp Ile Gln

450 455 460

Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val

465 470 475 480

Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Arg

485 490 495

Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys

500 505 510

Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val Pro Ser Arg

515 520 525

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser

530 535 540

Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Ser Tyr Asp

545 550 555 560

Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly

565 570 575

Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu

580 585 590

Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr

595 600 605

Thr Phe Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Lys

610 615 620

Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn

625 630 635 640

Tyr Ile Glu Lys Phe Lys Asn Arg Phe Thr Ile Ser Val Asp Lys Ala

645 650 655

Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

660 665 670

Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp

675 680 685

Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly

690 695 700

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

705 710 715 720

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

725 730 735

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

740 745 750

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

755 760 765

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

770 775 780

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

785 790 795 800

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

805 810 815

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

820 825 830

Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Ser Cys Ala Val

835 840 845

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

850 855 860

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

865 870 875 880

Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

885 890 895

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

900 905 910

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

915 920 925

<210> 251

<211> 1073

<212> PROTEIN

<213> Homo sapiens

<400> 251

Met Lys Thr Leu Leu Leu Asp Leu Ala Leu Trp Ser Leu Leu Phe Gln

1 5 10 15

Pro Gly Trp Leu Ser Phe Ser Ser Gln Val Ser Gln Asn Cys His Asn

20 25 30

Gly Ser Tyr Glu Ile Ser Val Leu Met Met Gly Asn Ser Ala Phe Ala

35 40 45

Glu Pro Leu Lys Asn Leu Glu Asp Ala Val Asn Glu Gly Leu Glu Ile

50 55 60

Val Arg Gly Arg Leu Gln Asn Ala Gly Leu Asn Val Thr Val Asn Ala

65 70 75 80

Thr Phe Met Tyr Ser Asp Gly Leu Ile His Asn Ser Gly Asp Cys Arg

85 90 95

Ser Ser Thr Cys Glu Gly Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala

100 105 110

Gln Arg Met Gly Cys Val Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr

115 120 125

Phe Gln Met Tyr Leu Asp Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala

130 135 140

Gly Ser Phe Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu

145 150 155 160

Met Ser Pro Ala Arg Lys Leu Met Tyr Phe Leu Val Asn Phe Trp Lys

165 170 175

Thr Asn Asp Leu Pro Phe Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val

180 185 190

Tyr Lys Asn Gly Thr Glu Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala

195 200 205

Leu Glu Ala Ser Val Ser Tyr Phe Ser His Glu Leu Gly Phe Lys Val

210 215 220

Val Leu Arg Gln Asp Lys Glu Phe Gln Asp Ile Leu Met Asp His Asn

225 230 235 240

Arg Lys Ser Asn Val Ile Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr

245 250 255

Lys Leu Lys Gly Asp Arg Ala Val Ala Glu Asp Ile Val Ile Ile Leu

260 265 270

Val Asp Leu Phe Asn Asp Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro

275 280 285

Asp Tyr Met Lys Asn Val Leu Val Leu Thr Leu Ser Pro Gly Asn Ser

290 295 300

Leu Leu Asn Ser Ser Phe Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp

305 310 315 320

Phe Ala Leu Ala Tyr Leu Asn Gly Ile Leu Leu Phe Gly His Met Leu

325 330 335

Lys Ile Phe Leu Glu Asn Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala

340 345 350

His Ala Phe Arg Asn Leu Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr

355 360 365

Leu Asp Asp Trp Gly Asp Val Asp Ser Thr Met Val Leu Leu Tyr Thr

370 375 380

Ser Val Asp Thr Lys Lys Tyr Lys Val Leu Leu Thr Tyr Asp Thr His

385 390 395 400

Val Asn Lys Thr Tyr Pro Val Asp Met Ser Pro Thr Phe Thr Trp Lys

405 410 415

Asn Ser Lys Leu Pro Asn Asp Ile Thr Gly Arg Gly Pro Gln Ile Leu

420 425 430

Met Ile Ala Val Phe Thr Leu Thr Gly Ala Val Val Leu Leu Leu Leu

435 440 445

Val Ala Leu Leu Met Leu Arg Lys Tyr Arg Lys Asp Tyr Glu Leu Arg

450 455 460

Gln Lys Lys Trp Ser His Ile Pro Pro Glu Asn Ile Phe Pro Leu Glu

465 470 475 480

Thr Asn Glu Thr Asn His Val Ser Leu Lys Ile Asp Asp Asp Lys Arg

485 490 495

Arg Asp Thr Ile Gln Arg Leu Arg Gln Cys Lys Tyr Asp Lys Lys Arg

500 505 510

Val Ile Leu Lys Asp Leu Lys His Asn Asp Gly Asn Phe Thr Glu Lys

515 520 525

Gln Lys Ile Glu Leu Asn Lys Leu Leu Gln Ile Asp Tyr Tyr Asn Leu

530 535 540

Thr Lys Phe Tyr Gly Thr Val Lys Leu Asp Thr Met Ile Phe Gly Val

545 550 555 560

Ile Glu Tyr Cys Glu Arg Gly Ser Leu Arg Glu Val Leu Asn Asp Thr

565 570 575

Ile Ser Tyr Pro Asp Gly Thr Phe Met Asp Trp Glu Phe Lys Ile Ser

580 585 590

Val Leu Tyr Asp Ile Ala Lys Gly Met Ser Tyr Leu His Ser Ser Lys

595 600 605

Thr Glu Val His Gly Arg Leu Lys Ser Thr Asn Cys Val Val Asp Ser

610 615 620

Arg Met Val Val Lys Ile Thr Asp Phe Gly Cys Asn Ser Ile Leu Pro

625 630 635 640

Pro Lys Lys Asp Leu Trp Thr Ala Pro Glu His Leu Arg Gln Ala Asn

645 650 655

Ile Ser Gln Lys Gly Asp Val Tyr Ser Tyr Gly Ile Ile Ala Gln Glu

660 665 670

Ile Ile Leu Arg Lys Glu Thr Phe Tyr Thr Leu Ser Cys Arg Asp Arg

675 680 685

Asn Glu Lys Ile Phe Arg Val Glu Asn Ser Asn Gly Met Lys Pro Phe

690 695 700

Arg Pro Asp Leu Phe Leu Glu Thr Ala Glu Glu Lys Glu Leu Glu Val

705 710 715 720

Tyr Leu Leu Val Lys Asn Cys Trp Glu Glu Asp Pro Glu Lys Arg Pro

725 730 735

Asp Phe Lys Lys Ile Glu Thr Thr Leu Ala Lys Ile Phe Gly Leu Phe

740 745 750

His Asp Gln Lys Asn Glu Ser Tyr Met Asp Thr Leu Ile Arg Arg Leu

755 760 765

Gln Leu Tyr Ser Arg Asn Leu Glu His Leu Val Glu Glu Arg Thr Gln

770 775 780

Leu Tyr Lys Ala Glu Arg Asp Arg Ala Asp Arg Leu Asn Phe Met Leu

785 790 795 800

Leu Pro Arg Leu Val Val Lys Ser Leu Lys Glu Lys Gly Phe Val Glu

805 810 815

Pro Glu Leu Tyr Glu Glu Val Thr Ile Tyr Phe Ser Asp Ile Val Gly

820 825 830

Phe Thr Thr Ile Cys Lys Tyr Ser Thr Pro Met Glu Val Val Asp Met

835 840 845

Leu Asn Asp Ile Tyr Lys Ser Phe Asp His Ile Val Asp His His Asp

850 855 860

Val Tyr Lys Val Glu Thr Ile Gly Asp Ala Tyr Met Val Ala Ser Gly

865 870 875 880

Leu Pro Lys Arg Asn Gly Asn Arg His Ala Ile Asp Ile Ala Lys Met

885 890 895

Ala Leu Glu Ile Leu Ser Phe Met Gly Thr Phe Glu Leu Glu His Leu

900 905 910

Pro Gly Leu Pro Ile Trp Ile Arg Ile Gly Val His Ser Gly Pro Cys

915 920 925

Ala Ala Gly Val Val Gly Ile Lys Met Pro Arg Tyr Cys Leu Phe Gly

930 935 940

Asp Thr Val Asn Thr Ala Ser Arg Met Glu Ser Thr Gly Leu Pro Leu

945 950 955 960

Arg Ile His Val Ser Gly Ser Thr Ile Ala Ile Leu Lys Arg Thr Glu

965 970 975

Cys Gln Phe Leu Tyr Glu Val Arg Gly Glu Thr Tyr Leu Lys Gly Arg

980 985 990

Gly Asn Glu Thr Thr Tyr Trp Leu Thr Gly Met Lys Asp Gln Lys Phe

995 1000 1005

Asn Leu Pro Thr Pro Pro Thr Val Glu Asn Gln Gln Arg Leu Gln

1010 1015 1020

Ala Glu Phe Ser Asp Met Ile Ala Asn Ser Leu Gln Lys Arg Gln

1025 1030 1035

Ala Ala Gly Ile Arg Ser Gln Lys Pro Arg Arg Val Ala Ser Tyr

1040 1045 1050

Lys Lys Gly Thr Leu Glu Tyr Leu Gln Leu Asn Thr Thr Asp Lys

1055 1060 1065

Glu Ser Thr Tyr Phe

1070

<210> 252

<211> 3850

<212> DNA

<213> Homo sapians

<400> 252

gaccagagag aagcgtgggg aagagtgggc tgagggactc cactagaggc tgtccatctg 60

gattccctgc ctccctagga gcccaacaga gcaaagcaag tgggcacaag gagtatggtt 120

ctaacgtgat tggggtcatg aagacgttgc tgttggactt ggctttgtgg tcactgctct 180

tccagcccgg gtggctgtcc tttagttccc aggtgagtca gaactgccac aatggcagct 240

atgaaatcag cgtcctgatg atgggcaact cagcctttgc agagcccctg aaaaacttgg 300

aagatgcggt gaatgagggg ctggaaatag tgagaggacg tctgcaaaat gctggcctaa 360

atgtgactgt gaacgctact ttcatgtatt cggatggtct gattcataac tcaggcgact 420

gccggagtag cacctgtgaa ggcctcgacc tactcaggaa aatttcaaat gcacaacgga 480

tgggctgtgt cctcataggg ccctcatgta catactccac cttccagatg taccttgaca 540

cagaattgag ctaccccatg atctcagctg gaagttttgg attgtcatgt gactataaag 600

aaaccttaac caggctgatg tctccagcta gaaagttgat gtacttcttg gttaactttt 660

ggaaaaccaa cgatctgccc ttcaaaactt attcctggag cacttcgtat gtttacaaga 720

atggtacaga aactgaggac tgtttctggt accttaatgc tctggaggct agcgtttcct 780

atttctccca cgaactcggc tttaaggtgg tgttaagaca agataaggag tttcaggata 840

tcttaatgga ccacaacagg aaaagcaatg tgattattat gtgtggtggt ccagagttcc 900

tctacaagct gaagggtgac cgagcagtgg ctgaagacat tgtcattatt ctagtggatc 960

ttttcaatga ccagtacttt gaggacaatg tcacagcccc tgactatatg aaaaatgtcc 1020

ttgttctgac gctgtctcct gggaattccc ttctaaatag ctctttctcc aggaatctat 1080

caccaacaaa acgagacttt gctcttgcct atttgaatgg aatcctgctc tttggacata 1140

tgctgaagat atttcttgaa aatggagaaa atattaccac ccccaaattt gctcatgctt 1200

tcaggaatct cacttttgaa gggtatgacg gtccagtgac cttggatgac tggggggatg 1260

ttgacagtac catggtgctt ctgtatacct ctgtggacac caagaaatac aaggttcttt 1320

tgacctatga tacccacgta aataagacct atcctgtgga tatgagcccc acattcactt 1380

ggaagaactc taaacttcct aatgatatta caggccgggg ccctcagatc ctgatgattg 1440

cagtcttcac cctcactgga gctgtggtgc tgctcctgct cgtcgctctc ctgatgctca 1500

gaaaatatag aaaagattat gaacttcgtc agaaaaaatg gtcccacatt cctcctgaaa 1560

atatctttcc tctggagacc aatgagacca atcatgttag cctcaagatc gatgatgaca 1620

aaagacgaga tacaatccag agactacgac agtgcaaata cgacaaaaag cgagtgattc 1680

tcaaagatct caagcacaat gatggtaatt tcactgaaaa acagaagata gaattgaaca 1740

agttgcttca gattgactat tacaacctga ccaagttcta cggcacagtg aaacttgata 1800

ccatgatctt cggggtgata gaatactgtg agagaggatc cctccgggaa gttttaaatg 1860

acacaatttc ctaccctgat ggcacattca tggattggga gtttaagatc tctgtcttgt 1920

atgacattgc taagggaatg tcatatctgc actccagtaa gacagaagtc catggtcgtc 1980

tgaaatctac caactgcgta gtggacagta gaatggtggt gaagatcact gattttggct 2040

gcaattccat tttacctcca aaaaaggacc tgtggacagc tccagagcac ctccgccaag 2100

ccaacatctc tcagaaagga gatgtgtaca gctatgggat catcgcacag gagatcatcc 2160

tgcggaaaga aaccttctac actttgagct gtcgggaccg gaatgagaag attttcagag 2220

tggaaaattc caatggaatg aaacccttcc gcccagattt attcttggaa acagcagagg 2280

aaaaagagct agaagtgtac ctacttgtaa aaaactgttg ggaggaagat ccagaaaaga 2340

gaccagattt caaaaaaatt gagactacac ttgccaagat atttggactt tttcatgacc 2400

aaaaaaatga aagctatatg gataccttga tccgacgtct acagctatat tctcgaaacc 2460

tggaacatct ggtagaggaa aggacacagc tgtacaaggc agagagggac agggctgaca 2520

gacttaactt tatgttgctt ccaaggctag tggtaaagtc tctgaaggag aaaggctttg 2580

tggagccgga actatatgag gaagttacaa tctacttcag tgacattgta ggtttcacta 2640

ctatctgcaa atacagcacc cccatggaag tggtggacat gcttaatgac atctataaga 2700

gttttgacca cattgttgat catcatgatg tctacaaggt ggaaaccatc ggtgatgcgt 2760

acatggtggc tagtggtttg cctaagagaa atggcaatcg gcatgcaata gacattgcca 2820

agatggcctt ggaaatcctc agcttcatgg ggacctttga gctggagcat cttcctggcc 2880

tcccaatatg gattcgcatt ggagttcact ctggtccctg tgctgctgga gttgtgggaa 2940

tcaagatgcc tcgttattgt ctatttggag atacggtcaa cacagcctct aggatggaat 3000

ccactggcct ccctttgaga attcacgtga gtggctccac catagccatc ctgaagagaa 3060

ctgagtgcca gttcctttat gaagtgagag gagaaacata cttaaaggga agaggaaatg 3120

agactaccta ctggctgact gggatgaagg accagaaatt caacctgcca acccctccta 3180

ctgtggagaa tcaacagcgt ttgcaagcag aattttcaga catgattgcc aactctttac 3240

agaaaagaca ggcagcaggg ataagaagcc aaaaacccag acgggtagcc agctataaaa 3300

aaggcactct ggaatacttg cagctgaata ccacagacaa ggagagcacc tatttttaaa 3360

cctaaatgag gtataaggac tcacacaaat taaaatacag ctgcactgag gcagcgacct 3420

caagtgtcct gaaagcttac attttcctga gacctcaatg aagcagaaat gtacttaggc 3480

ttggctgccc tgtctggaac atggactttc ttgcatgaat cagatgtgtg ttctcagtga 3540

aataactacc ttccactctg gaaccttatt ccagcagttg ttccagggag cttctacctg 3600

gaaaagaaaa gaaatgaata gactatctag aacttgagaa gattttattc ttatttcatt 3660

tattttttgt ttgtttattt ttatcgtttt tgtttactgg ctttccttct gtattcataa 3720

gattttttaa attgtcataa ttatatttta aatacccatc ttcattaaag tatatttaac 3780

tcataatttt tgcagaaaat atgctatata ttaggcaaga ataaaagcta aaggtttccc 3840

aaaaaaaaaa 3850

<210> 253

<211> 1050

<212> PROTEIN

<213> Macaca fascicularis

<400> 253

Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val

1 5 10 15

Leu Met Met Asp Asn Ser Ala Phe Ala Glu Pro Leu Glu Asn Val Glu

20 25 30

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

35 40 45

Ala Gly Leu Asn Val Thr Val Asn Ala Ser Phe Met Tyr Ser Asp Gly

50 55 60

Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu

65 70 75 80

Asp Leu Leu Arg Lys Ile Ser Asn Ala Lys Arg Met Gly Cys Val Leu

85 90 95

Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr

100 105 110

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

115 120 125

Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu

130 135 140

Thr Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys

145 150 155 160

Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Ser

165 170 175

Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr

180 185 190

Phe Ser His Glu Leu Ser Phe Lys Leu Val Leu Arg Gln Asp Lys Glu

195 200 205

Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Val

210 215 220

Met Cys Gly Asp Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala

225 230 235 240

Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln

245 250 255

Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu

260 265 270

Val Leu Thr Gln Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

275 280 285

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

290 295 300

Gly Ile Leu Leu Phe Gly His Met Leu Lys Thr Phe Leu Glu Asn Gly

305 310 315 320

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

325 330 335

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

340 345 350

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

355 360 365

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Gln Thr Asn Pro Val

370 375 380

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

385 390 395 400

Ile Thr Asp Arg Gly Pro Gln Ile Leu Met Ile Ala Val Phe Thr Leu

405 410 415

Thr Gly Ala Val Val Leu Leu Leu Leu Val Ala Leu Leu Met Leu Arg

420 425 430

Lys Tyr Lys Lys Asp Tyr Glu Leu Arg Gln Lys Lys Trp Ser His Ile

435 440 445

Pro Pro Glu Asn Ile Phe Pro Leu Glu Thr Asn Glu Thr Asn His Val

450 455 460

Ser Leu Lys Ile Asp Asp Asp Lys Arg Arg Asp Thr Ile Gln Arg Leu

465 470 475 480

Arg Gln Cys Lys Tyr Asp Lys Lys Arg Val Ile Leu Lys Asp Leu Lys

485 490 495

His Asn Asp Gly Asn Phe Thr Glu Lys Gln Lys Ile Glu Leu Asn Lys

500 505 510

Leu Leu Gln Ile Asp Tyr Tyr Asn Leu Thr Lys Phe Tyr Gly Thr Val

515 520 525

Lys Leu Asp Thr Met Ile Phe Gly Val Ile Glu Tyr Cys Glu Arg Gly

530 535 540

Ser Leu Arg Glu Val Leu Asn Asp Thr Ile Ser Tyr Pro Asp Gly Thr

545 550 555 560

Phe Met Asp Trp Glu Phe Lys Ile Ser Val Leu Tyr Asp Ile Ala Lys

565 570 575

Gly Met Ser Tyr Leu His Ser Ser Lys Thr Glu Val His Gly Arg Leu

580 585 590

Lys Ser Thr Asn Cys Val Val Asp Ser Arg Met Val Val Lys Ile Thr

595 600 605

Asp Phe Gly Cys Asn Ser Ile Leu Pro Pro Lys Lys Asp Leu Trp Thr

610 615 620

Ala Pro Glu His Leu Arg Gln Ala Asn Val Ser Gln Lys Gly Asp Val

625 630 635 640

Tyr Ser Tyr Gly Ile Ile Ala Gln Glu Ile Ile Leu Arg Lys Glu Thr

645 650 655

Phe Tyr Thr Ser Ser Cys Arg Asp Arg Asn Glu Lys Ile Phe Arg Val

660 665 670

Glu Asn Ser Asn Gly Met Lys Pro Phe Arg Pro Asp Leu Phe Leu Glu

675 680 685

Thr Ala Glu Glu Lys Glu Leu Glu Val Tyr Leu Leu Val Lys Ser Cys

690 695 700

Trp Glu Glu Asp Pro Glu Lys Arg Pro Asp Phe Lys Lys Ile Glu Thr

705 710 715 720

Thr Leu Ala Lys Ile Phe Gly Leu Phe His Asp Gln Lys Asn Glu Ser

725 730 735

Tyr Met Asp Thr Leu Ile Arg Arg Leu Gln Leu Tyr Ser Arg Asn Leu

740 745 750

Glu His Leu Val Glu Glu Arg Thr Gln Leu Tyr Lys Ala Glu Arg Asp

755 760 765

Arg Ala Asp Arg Leu Asn Phe Met Leu Leu Pro Arg Leu Val Val Lys

770 775 780

Ser Leu Lys Glu Lys Gly Phe Val Glu Pro Glu Leu Tyr Glu Glu Val

785 790 795 800

Thr Ile Tyr Phe Ser Asp Ile Val Gly Phe Thr Thr Ile Cys Lys Tyr

805 810 815

Ser Thr Pro Met Glu Val Val Asp Met Leu Asn Asp Ile Tyr Lys Ser

820 825 830

Phe Asp His Ile Val Asp His His Asp Val Tyr Lys Val Glu Thr Ile

835 840 845

Gly Asp Ala Tyr Met Val Ala Ser Gly Leu Pro Lys Arg Asn Gly Asn

850 855 860

Arg His Ala Ile Asp Ile Ala Lys Met Ala Leu Glu Ile Leu Ser Phe

865 870 875 880

Met Gly Thr Phe Glu Leu Glu His Leu Pro Gly Leu Pro Ile Trp Ile

885 890 895

Arg Ile Gly Val His Ser Gly Pro Cys Ala Ala Gly Val Val Gly Ile

900 905 910

Lys Met Pro Arg Tyr Cys Leu Phe Gly Asp Thr Val Asn Thr Ala Ser

915 920 925

Arg Met Glu Ser Thr Gly Leu Pro Leu Arg Ile His Val Ser Gly Ser

930 935 940

Thr Ile Ala Ile Leu Lys Arg Thr Glu Cys Gln Phe Leu Tyr Glu Val

945 950 955 960

Arg Gly Glu Thr Tyr Leu Lys Gly Arg Gly Asn Glu Thr Thr Tyr Trp

965 970 975

Leu Thr Gly Met Lys Asp Gln Lys Phe Asn Leu Pro Thr Pro Pro Thr

980 985 990

Val Glu Asn Gln Gln Arg Leu Gln Ala Glu Phe Ser Asp Met Ile Ala

995 1000 1005

Asn Ser Leu Gln Lys Arg Gln Ala Ala Gly Ile Arg Ser Gln Lys

1010 1015 1020

Pro Arg Arg Val Ala Ser Tyr Lys Lys Gly Thr Leu Glu Tyr Leu

1025 1030 1035

Gln Leu Asn Thr Thr Asp Lys Glu Ser Thr Tyr Phe

1040 1045 1050

<210> 254

<211> 3150

<212> DNA

<213> Macaca fascicularis

<400> 254

tcacaggtga gtcagaactg ccacaatggc agctatgaaa tcagcgtcct gatgatggac 60

aactcagcct ttgcagagcc cctggaaaac gtggaagatg cggtgaatga ggggctggaa 120

atagtgagag gacgtctgca aaacgctggc ctaaatgtga ctgtgaatgc ttctttcatg 180

tattcggatg gtctgattca taactccggc gactgccgga gcagcacctg tgaaggcctt 240

gacctactca ggaaaatttc aaatgcaaaa cggatgggct gtgtcctcat ggggccctca 300

tgtacatact ccaccttcca gatgtacctt gacacagaat tgagctaccc catgatctca 360

gctggaagtt ttggattgtc atgtgactat aaagaaacct taaccaggct gatgtctcca 420

gctagaaagt tgacatactt cttggttaac ttttggaaaa ccaatgatct acccttcaaa 480

acttattcct ggagcacttc gtatgtttac aagaatggta cggagtccga ggactgtttc 540

tggtacctta acgctctgga ggccagtgtt tcctatttct cccacgaact cagttttaag 600

ttggtgttaa gacaagataa ggagtttcag gatatcttaa tggaccacaa caggaaaagc 660

aatgtgattg ttatgtgtgg tgatccagag ttcctctaca agttgaaggg tgaccgagca 720

gtggctgaag acattgtcat tattctagtg gatcttttca atgaccagta ctttgaggac 780

aatgtcacag cccctgacta tatgaaaaat gtccttgttc tgacgcagtc tcctggcaat 840

tccctcctaa atagctcttt ctccaggaat ctatccccaa caaaacgaga ctttgctctt 900

gcctatttga atggaatcct gctctttgga catatgctaa agacatttct tgaaaatgga 960

gaaaatatta ccacccccaa atttgctcat gctttcagga atctcacttt tgaagggtat 1020

gacggtccag tgaccttgga tgactgggg gatgtggaca gtaccatggt gcttctgtat 1080

acgtctgtgg acaccaagaa atacaaggtt cttttgacct atgataccca cgtaaatcag 1140

accaaccctg tggatatgag ccccacattc acttggaaga actctaaact tcctaatgat 1200

attacagacc ggggccctca gatcctgatg attgcagtct tcaccctcac cggagctgtg 1260

gtgctgctcc tgcttgtcgc tctcctgatg ctcagaaaat ataaaaaaga ttatgaactt 1320

cgtcagaaaa aatggtccca cattcctcct gaaaatatct ttcctctgga gaccaatgag 1380

accaatcacg ttagcctgaa gatcgatgat gacaaaagac gagatacaat cccagacta 1440

cgacagtgca aatacgacaa aaagcgagtg attctcaaag atctcaagca caatgatggt 1500

aatttcactg aaaaacagaa gatagaattg aacaagttgc ttcagattga ctattacaac 1560

ctgaccaagt tctatggcac cgtgaaactt gataccatga tcttcggggt gatagaatac 1620

tgtgagagag gatccctccg ggaagtttta aatgacacaa tttcctaccc tgatggcaca 1680

ttcatggatt gggagtttaa gatctctgtc ctgtatgaca ttgctaaggg aatgtcatat 1740

ctgcactcca gtaagacaga agtccatggt cgtctgaaat ctaccaactg cgtagtggac 1800

agtagaatgg tggtgaagat cactgatttt ggctgcaatt ccattttacc tccaaaaaaa 1860

gacctgtgga cagctccaga gcacctccgc caagccaacg tctctcagaa aggagatgtg 1920

tacagctacg ggatcatcgc acaggagatc atcctgcgga aagaaacctt ctacacttcg 1980

agctgtcgag accggaacga gaagattttc agagtggaaa attccaatgg aatgaaaccc 2040

ttccgtccag atttattctt ggaaacggca gaggaaaaag agctagaagt gtacctactt 2100

gtaaaaagct gttgggaaga agatccagaa aagagaccag atttcaaaaa aattgagact 2160

acacttgcca agatatttgg actttttcat gaccaaaaaa atgaaagcta tatggatacc 2220

ttgatccgac gtctacagct atattctcga aacctggaac atctggtaga ggaaaggaca 2280

cagctataca aggcagagag ggacagggct gacagactta actttatgtt gcttccaagg 2340

ctagtggtaa agtctctgaa ggagaaaggc tttgtagagc cggaactata tgaggaagtt 2400

acaatctact tcagtgacat tgtaggtttc actactatct gcaaatacag cacccccatg 2460

gaagtggtgg acatgcttaa tgacatctat aagagttttg accacattgt tgatcatcat 2520

gatgtctaca aggtggaaac cattggtgat gcctacatgg tggctagtgg tttgcctaag 2580

agaaatggca atcggcatgc aatagacatt gccaagatgg ccttggaaat cctcagcttc 2640

atggggacct ttgagctgga gcatcttcct ggcctcccaa tatggattcg cattggcgtt 2700

cactctggtc cctgcgctgc tggagttgtg ggaatcaaga tgcctcgtta ttgtctattt 2760

ggagatacag tcaacacagc ctctaggatg gaatccactg gcctcccttt gaggattcat 2820

gtgagtggct ccaccatagc cattctgaag agaactgagt gccagttcct gtatgaagtg 2880

agaggagaaa cgtacttaaa gggaagagga aatgagacta cctactggct gaccgggatg 2940

aaggaccaga aattcaacct gccaacccct cctactgtgg agaatcaaca gcgtttgcaa 3000

gcagaatttt cagacatgat tgccaactct ttacagaaaa gacaggcggc agggataaga 3060

agccaaaaac cgacgagt agccagctat aaaaaaggca ctctggaata cttgcaactg 3120

aataccacgg acaaggag cacctatttt 3150

<210> 255

<211> 1072

<212> PROTEIN

<213> Mus musculus

<400> 255

Met Thr Ser Leu Leu Gly Leu Ala Val Arg Leu Leu Leu Phe Gln Pro

1 5 10 15

Ala Leu Met Val Phe Trp Ala Ser Gln Val Arg Gln Asn Cys Arg Asn

20 25 30

Gly Ser Tyr Glu Ile Ser Val Leu Met Met Asp Asn Ser Ala Tyr Lys

35 40 45

Glu Pro Met Gln Asn Leu Arg Glu Ala Val Glu Glu Gly Leu Asp Ile

50 55 60

Val Arg Lys Arg Leu Arg Glu Ala Asp Leu Asn Val Thr Val Asn Ala

65 70 75 80

Thr Phe Ile Tyr Ser Asp Gly Leu Ile His Lys Ser Gly Asp Cys Arg

85 90 95

Ser Ser Thr Cys Glu Gly Leu Asp Leu Leu Arg Glu Ile Thr Arg Asp

100 105 110

His Lys Met Gly Cys Ala Leu Met Gly Pro Ser Cys Thr Tyr Ser Thr

115 120 125

Phe Gln Met Tyr Leu Asp Thr Glu Leu Asn Tyr Pro Met Ile Ser Ala

130 135 140

Gly Ser Tyr Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Ile

145 150 155 160

Leu Pro Pro Ala Arg Lys Leu Met Tyr Phe Leu Val Asp Phe Trp Lys

165 170 175

Val Asn Asn Ala Ser Phe Lys Pro Phe Ser Trp Asn Ser Ser Tyr Val

180 185 190

Tyr Lys Asn Gly Ser Glu Pro Glu Asp Cys Phe Trp Tyr Leu Asn Ala

195 200 205

Leu Glu Ala Gly Val Ser Tyr Phe Ser Glu Val Leu Asn Phe Lys Asp

210 215 220

Val Leu Arg Arg Ser Glu Gln Phe Gln Glu Ile Leu Thr Gly His Asn

225 230 235 240

Arg Lys Ser Asn Val Ile Val Met Cys Gly Thr Pro Glu Ser Phe Tyr

245 250 255

Asp Val Lys Gly Asp Leu Gln Val Ala Glu Asp Thr Val Val Ile Leu

260 265 270

Val Asp Leu Phe Ser Asn His Tyr Phe Glu Glu Asn Thr Thr Ala Pro

275 280 285

Glu Tyr Met Asp Asn Val Leu Val Leu Thr Leu Pro Ser Glu Gln Ser

290 295 300

Thr Ser Asn Thr Ser Val Ala Glu Arg Phe Ser Ser Gly Arg Ser Asp

305 310 315 320

Phe Ser Leu Ala Tyr Leu Glu Gly Thr Leu Leu Phe Gly His Met Leu

325 330 335

Gln Thr Phe Leu Glu Asn Gly Glu Asn Val Thr Gly Pro Lys Phe Ala

340 345 350

Arg Ala Phe Arg Asn Leu Thr Phe Gln Gly Phe Ala Gly Pro Val Thr

355 360 365

Leu Asp Asp Ser Gly Asp Ile Asp Asn Ile Met Ser Leu Leu Tyr Val

370 375 380

Ser Leu Asp Thr Arg Lys Tyr Lys Val Leu Met Lys Tyr Asp Thr His

385 390 395 400

Lys Asn Lys Thr Ile Pro Val Ala Glu Asn Pro Asn Phe Ile Trp Lys

405 410 415

Asn His Lys Leu Pro Asn Asp Val Pro Gly Leu Gly Pro Gln Ile Leu

420 425 430

Met Ile Ala Val Phe Thr Leu Thr Gly Ile Leu Val Val Leu Leu Leu

435 440 445

Ile Ala Leu Leu Val Leu Arg Lys Tyr Arg Arg Asp His Ala Leu Arg

450 455 460

Gln Lys Lys Trp Ser His Ile Pro Ser Glu Asn Ile Phe Pro Leu Glu

465 470 475 480

Thr Asn Glu Thr Asn His Ile Ser Leu Lys Ile Asp Asp Asp Arg Arg

485 490 495

Arg Asp Thr Ile Gln Arg Val Arg Gln Cys Lys Tyr Asp Lys Lys Lys

500 505 510

Val Ile Leu Lys Asp Leu Lys His Ser Asp Gly Asn Phe Ser Glu Lys

515 520 525

Gln Lys Ile Asp Leu Asn Lys Leu Leu Gln Ser Asp Tyr Tyr Asn Leu

530 535 540

Thr Lys Phe Tyr Gly Thr Val Lys Leu Asp Thr Arg Ile Phe Gly Val

545 550 555 560

Val Glu Tyr Cys Glu Arg Gly Ser Leu Arg Glu Val Leu Asn Asp Thr

565 570 575

Ile Ser Tyr Pro Asp Gly Thr Phe Met Asp Trp Glu Phe Lys Ile Ser

580 585 590

Val Leu Asn Asp Ile Ala Lys Gly Met Ser Tyr Leu His Ser Ser Lys

595 600 605

Ile Glu Val His Gly Arg Leu Lys Ser Thr Asn Cys Val Val Asp Ser

610 615 620

Arg Met Val Val Lys Ile Thr Asp Phe Gly Cys Asn Ser Ile Leu Pro

625 630 635 640

Pro Lys Lys Asp Leu Trp Thr Ala Pro Glu His Leu Arg Gln Ala Thr

645 650 655

Ile Ser Gln Lys Gly Asp Val Tyr Ser Phe Ala Ile Ile Ala Gln Glu

660 665 670

Ile Ile Leu Arg Lys Glu Thr Phe Tyr Thr Leu Ser Cys Arg Asp His

675 680 685

Asn Glu Lys Ile Phe Arg Val Glu Asn Ser Tyr Gly Lys Pro Phe Arg

690 695 700

Pro Asp Leu Phe Leu Glu Thr Ala Asp Glu Lys Glu Leu Glu Val Tyr

705 710 715 720

Leu Leu Val Lys Ser Cys Trp Glu Glu Asp Pro Glu Lys Arg Pro Asp

725 730 735

Phe Lys Lys Ile Glu Ser Thr Leu Ala Lys Ile Phe Gly Leu Phe His

740 745 750

Asp Gln Lys Asn Glu Ser Tyr Met Asp Thr Leu Ile Arg Arg Leu Gln

755 760 765

Leu Tyr Ser Arg Asn Leu Glu His Leu Val Glu Glu Arg Thr Gln Leu

770 775 780

Tyr Lys Ala Glu Arg Asp Arg Ala Asp His Leu Asn Phe Met Leu Leu

785 790 795 800

Pro Arg Leu Val Val Lys Ser Leu Lys Glu Lys Gly Ile Val Glu Pro

805 810 815

Glu Leu Tyr Glu Glu Val Thr Ile Tyr Phe Ser Asp Ile Val Gly Phe

820 825 830

Thr Thr Ile Cys Lys Tyr Ser Thr Pro Met Glu Val Val Asp Met Leu

835 840 845

Asn Asp Ile Tyr Lys Ser Phe Asp Gln Ile Val Asp His His Asp Val

850 855 860

Tyr Lys Val Glu Thr Ile Gly Asp Ala Tyr Val Val Ala Ser Gly Leu

865 870 875 880

Pro Met Arg Asn Gly Asn Arg His Ala Val Asp Ile Ser Lys Met Ala

885 890 895

Leu Asp Ile Leu Ser Phe Ile Gly Thr Phe Glu Leu Glu His Leu Pro

900 905 910

Gly Leu Pro Val Trp Ile Arg Ile Gly Val His Ser Gly Pro Cys Ala

915 920 925

Ala Gly Val Val Gly Ile Lys Met Pro Arg Tyr Cys Leu Phe Gly Asp

930 935 940

Thr Val Asn Thr Ala Ser Arg Met Glu Ser Thr Gly Leu Pro Leu Arg

945 950 955 960

Ile His Met Ser Ser Ser Thr Ile Thr Ile Leu Lys Arg Thr Asp Cys

965 970 975

Gln Phe Leu Tyr Glu Val Arg Gly Glu Thr Tyr Leu Lys Gly Arg Gly

980 985 990

Thr Glu Thr Thr Tyr Trp Leu Thr Gly Met Lys Asp Gln Glu Tyr Asn

995 1000 1005

Leu Pro Ser Pro Pro Thr Val Glu Asn Gln Gln Arg Leu Gln Thr

1010 1015 1020

Glu Phe Ser Asp Met Ile Val Ser Ala Leu Gln Lys Arg Gln Ala

1025 1030 1035

Ser Gly Lys Lys Ser Arg Arg Pro Thr Arg Val Ala Ser Tyr Lys

1040 1045 1050

Lys Gly Phe Leu Glu Tyr Met Gln Leu Asn Asn Ser Asp His Asp

1055 1060 1065

Ser Thr Tyr Phe

1070

<210> 256

<211> 3951

<212> DNA

<213> Mus musculus

<400> 256

gaccagtgtg gcaagaccag aaaggtgcgt ggggaagaga agaccaaggg actctgctag 60

cgactctcca gaggggctcc ctgtgtctct aaaagcgagc aatccagggg ggcatggtgc 120

tacggtgagc caggtcatga cgtcactgct gggcttggct gtgcggttac tgctcttcca 180

gcccgcgctg atggtgttct gggcctctca ggtgaggcag aactgccgca atggcagcta 240

cgagatcagc gtcctgatga tggacaactc agcctacaaa gaacctatgc aaaacctgag 300

ggaggctgtg gaggaaggac tggacatagt gcgaaagcgc ctgcgtgaag ccgacctaaa 360

tgtgactgtg aacgcgactt tcatctactc cgacggtctg attcataagt caggtgactg 420

ccggagcagc acctgtgaag gccttgacct actcagggag attacaagag atcataagat 480

gggctgcgcc ctcatggggc cctcgtgcac gtattccacc ttccagatgt acctcgacac 540

agagttgaac tatcccatga tttccgctgg aagttatgga ttgtcctgtg actataagga 600

aaccctaacc aggatcctgc ctccagccag gaagctgatg tacttcttgg tcgatttctg 660

gaaagtcaac aatgcatctt tcaaaccctt ttcctggaac tcttcgtatg tttacaagaa 720

tggatcggaa cctgaagatt gtttctggta cctcaatgct ctggaggctg gggtgtccta 780

tttttctgag gtgctcaact tcaaggatgt actgagacgc agcgaacagt tccaggaaat 840

cttaacaggc cataacagaa agagcaatgt gattgttatg tgtggcacgc cagaaagctt 900

ctatgatgtg aaaggtgacc tccaagtggc tgaagatact gttgtcatcc tggtagatct 960

gttcagtaac cattactttg aggagaacac cacagctcct gagtatatgg acaatgtcct 1020

cgtcctgacg ctgccgtctg aacagtccac ctcaaacacc tctgtcgccg agaggttttc 1080

atcggggaga agtgactttt ctctcgctta cttggaggga accttgctat ttggacacat 1140

gctgcagacg tttcttgaaa atggagaaaa tgtcacgggt cccaagtttg ctcgtgcatt 1200

caggaatctc acttttcaag gctttgcagg acctgtgact ctggatgaca gtggggacat 1260

tgacaacatt atgtcccttc tgtatgtgtc tctggatacc aggaaataca aggttcttat 1320

gaagtatgac acccacaaaa acaaaactat tccggtggct gagaacccca acttcatctg 1380

gaagaaccac aagctcccca atgacgttcc tgggctgggc cctcaaatcc tgatgattgc 1440

cgtcttcacg ctcacgggga tcctggtagt tctgctgctg attgccctcc tcgtgctgag 1500

aaaatacaga agagatcatg cacttcgaca gaagaaatgg tcccacattc cttctgaaaa 1560

catctttcct ctggagacca acgagaccaa ccacatcagc ctgaagattg acgatgacag 1620

gagacgagac acaatccaga gagtgcgaca gtgcaaatac gacaagaaga aagtgattct 1680

gaaagacctc aagcacagcg acgggaactt cagtgagaag cagaagatag acctgaacaa 1740

gttgctgcag tctgactact acaacctgac taagttctac ggcaccgtga agctggacac 1800

caggatcttt ggggtggttg agtactgcga gaggggatcc ctccgggaag tgttaaacga 1860

cacaatttcc taccctgacg gcacgttcat ggattgggag tttaagatct ctgtcttaaa 1920

tgacatcgct aaggggatgt cctacctgca ctccagtaag attgaagtcc acgggcgtct 1980

caaatccacc aactgcgtgg tggacagccg catggtggtg aagatcaccg actttgggtg 2040

caattccatc ctgcctccaa aaaaagacct gtggacggcc ccggagcacc tgcgccaggc 2100

caccatctct cagaaaggag acgtgtacag cttcgccatc attgcccagg agatcatcct 2160

ccgtaaggag actttttaca cgctgagctg tcgggatcac aatgagaaga ttttcagagt 2220

ggaaaattca tacgggaaac ctttccgccc agacctcttc ctggagactg cagatgagaa 2280

ggagctggag gtctatctac ttgtcaaaag ctgttgggag gaggatccag aaaagaggcc 2340

agatttcaag aaaatcgaga gcacactggc caagatattt ggccttttcc atgaccagaa 2400

aaacgagtct tacatggaca ccttgatccg acgtctccag ctgtactctc gaaacctgga 2460

acatctggtg gaggaaagga ctcagctgta caaggcggag agggacaggg ctgaccacct 2520

taacttcatg ctcctcccac ggctggtggt aaagtcactg aaggagaaag gcatcgtgga 2580

gccagagctg tacgaagaag tcacaatcta cttcagtgac attgtgggct tcaccaccat 2640

ctgcaagtat agcacgccca tggaggtggt ggacatgctc aacgacatct acaagagctt 2700

tgaccagatt gtggaccacc atgacgtcta caaggtagaa accatcggtg acgcctacgt 2760

ggtggccagc ggtctgccta tgagaaacgg caaccgacac gcggtagaca tttccaagat 2820

ggccttggac atcctcagct tcatagggac ctttgagttg gagcatctcc ctggcctccc 2880

cgtgtggatc cgcattggag ttcattctgg gccctgcgct gctggtgttg tggggatcaa 2940

gatgcctcgc tattgcctgt ttggagacac tgtcaacact gcctccagga tggaatccac 3000

cggcctcccc ttgaggattc acatgagcag ctccaccata accatcctga agagaacgga 3060

ttgccagttc ctgtatgaag tgaggggaga aacctactta aagggaagag ggacagagac 3120

cacatactgg ctgactggga tgaaggacca agaatacaac ctgccatccc caccgacagt 3180

ggagaaccaa cagcgtctgc agactgagtt ctcagacatg atcgttagcg ccttacagaa 3240

aagacaggcc tcgggcaaga agagccggag gcccactcgg gtggccagct acaagaaagg 3300

ctttctggaa tacatgcagc tgaacaattc agaccacgat agcacctatt tttagaccaa 3360

gtgaggtctg agaactgaca gtagcaacct cctatatcat gaatctgtat tttccagaga 3420

cctcaacaac atagacaagc acttagcctc agtgccctga ctggaacgta gaaccaaccc 3480

ctcaagtcat gtgggtctga ttttgggttg gttggttggt tggttggttg gttttggttt 3540

tgttgagaca gagtttcgtg tatcccaagc tggtctcaaa ctcactgtgt agcagcagat 3600

gactttggac ttctaagatc atccgtgtgt gttcctgaca gtgtgatgag tgtatacgtc 3660

agagccctgt cccacagttc tccatggagc atcaacctga atggagggagc gaggaggggg 3720

aatgtcctgt gctgtacaga acttggggtt ttgttctaat tttatctctg gtttggtttt 3780

gttttctggc tccttccctc tctatgtgtg agagaagttt ttaaattgtc tgaattgtat 3840

gctaagtagc ttatctacaa gaaagtgtgt ttaactagtg atttttgcag aaaccatgct 3900

ggatattagg taaaaaataa aagtgtttag agtctaaaaa aaaaaaaaaa a 3951

<210> 257

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 257

Thr Tyr Trp Met Gln

15

<210> 258

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 258

Lys Tyr Trp Met Gln

15

<210> 259

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 259

Ser Tyr Trp Met His

15

<210> 260

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 260

Asp Thr Tyr Ile His

15

<210> 261

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 261

Asp Tyr Ile Met Leu

15

<210> 262

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 262

Tyr Pro Gly Asp Gly Met

15

<210> 263

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 263

Tyr Pro Gly Asp Gly Phe

15

<210> 264

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 264

Lys Pro Ser Asn Gly Leu

15

<210> 265

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 265

Asp Pro Ala Asn Gly Asn

15

<210> 266

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 266

Asn Pro Tyr Tyr Gly Ser

15

<210> 267

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 267

Lys Pro Ser Asn Glu Leu

15

<210> 268

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 268

Asp Tyr Tyr Met Thr

15

<210> 269

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 269

Arg Asn Arg Ala Arg Gly Tyr Thr

15

<210> 270

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 270

Arg Asn Gln Ala Arg Gly Tyr Thr

15

<210> 271

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 271

Ser Tyr Trp Met Ser

15

<210> 272

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 272

Lys Gln Asp Gly Ser Glu

15

<210> 273

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 273

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr Thr Ser Asp His Asn Pro

50 55 60

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr Val Leu Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 274

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 274

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Asp Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gly Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Asp Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 275

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 275

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Glu Ser Leu Phe Asn Val

20 25 30

Arg Ser Gly Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Asp Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 276

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 276

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gly Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Asp Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 277

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 277

Gly Phe Thr Phe Ser Asp Tyr

15

<210> 278

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 278

Thr Ser Asp Gln Ser Leu Phe Asn Val Arg Ser Gly Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 279

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 279

Thr Ser Ser Glu Ser Leu Phe Asn Val Arg Ser Gly Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 280

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 280

Thr Ser Ser Gln Ser Leu Phe Asn Val Arg Ser Gly Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 281

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 281

Trp Ala Ser Asp Arg Glu Ser

15

<210> 282

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 282

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 283

<211> 465

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 283

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Asp Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gly Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Asp Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser

115 120 125

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

130 135 140

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln

145 150 155 160

Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn

165 170 175

Glu Leu Thr Asn Val His Glu Lys Phe Lys Asp Arg Phe Thr Ile Ser

180 185 190

Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr

210 215 220

Glu Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr

225 230 235 240

Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

370 375 380

Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly

465

<210> 284

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 284

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 285

<211> 465

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 285

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gly Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Asp Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser

115 120 125

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

130 135 140

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln

145 150 155 160

Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn

165 170 175

Glu Leu Thr Asn Val His Glu Lys Phe Lys Asp Arg Phe Thr Ile Ser

180 185 190

Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr

210 215 220

Glu Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr

225 230 235 240

Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

370 375 380

Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly

465

<210> 286

<211> 461

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 286

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala Pro Ser Val Phe

245 250 255

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

260 265 270

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

275 280 285

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

290 295 300

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

305 310 315 320

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

325 330 335

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

340 345 350

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu Pro Pro

355 360 365

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val

370 375 380

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

385 390 395 400

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

405 410 415

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

420 425 430

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

435 440 445

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455 460

<210> 287

<211> 465

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 287

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Glu Ser Leu Phe Asn Val

20 25 30

Arg Ser Gly Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Asp Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser

115 120 125

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

130 135 140

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln

145 150 155 160

Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn

165 170 175

Glu Leu Thr Asn Val His Glu Lys Phe Lys Asp Arg Phe Thr Ile Ser

180 185 190

Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr

210 215 220

Glu Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr

225 230 235 240

Val Ser Ser Gly Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Ala

245 250 255

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

260 265 270

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

275 280 285

Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His

290 295 300

Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg

305 310 315 320

Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys

325 330 335

Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu

340 345 350

Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr

355 360 365

Thr Leu Pro Pro Cys Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu

370 375 380

Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp

385 390 395 400

Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val

405 410 415

Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp

420 425 430

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

435 440 445

Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro

450 455 460

Gly

465

<210> 288

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 288

gacatccaga tgacccagtc cccctcttct ctgtctgcct ctgtgggcga cagagtgacc 60

atcacctgca caagcgacca gtcactgttt aatgtccgca gcggcaaaaa ctatcttgcg 120

tggtatcagc agaagcctgg caaggctccc aagctgctga tctactgggc cagtgaccga 180

gaatccggcg tgccttccag attctccggc tctggctctg gcaccgattt caccctgacc 240

atctcctccc tccagcctga ggatttcgcc acctactact gcaaacagtc ttacgacctt 300

ttcacttttg gcggcggaac aaaggtggag atcaag 336

<210> 289

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 289

gaagtgcagc ttgttgaatc tggcggcggt ttggttcagc ccggtggatc actgcgactc 60

agttgcgcag ctagcggctt caccttttct gattactaca tgacatgggt acgacaggcg 120

ccaggcaagg gtttggaatg ggtagcattc atacgcaatc aggcacgcgg gtacacttca 180

gaccacaatc cctcagtaaa aggaagattt accatctcaa gagacaatgc caaaaattca 240

ctctacctgc aaatgaactc acttcgcgcc gaggataccg ccgtgtatta ctgtgccaga 300

gacagaccat cttattacgt gctggactat tggggacagg gcactacagt caccgtcagc 360

tct 363

<210> 290

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 290

gacatccaga tgacccagtc cccctcttct ctgtctgcct ctgtgggcga cagagtgacc 60

atcacctgca caagctcaga gtcactgttt aatgtccgca gcggcaaaaa ctatcttgcg 120

tggtatcagc agaagcctgg caaggctccc aagctgctga tctactgggc cagtgaccga 180

gaatccggcg tgccttccag attctccggc tctggctctg gcaccgattt caccctgacc 240

atctcctccc tccagcctga ggatttcgcc acctactact gcaaacagtc ttacgacctt 300

ttcacttttg gcggcggaac aaaggtggag atcaag 336

<210> 291

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 291

gaagtgcagc ttgttgaatc tggcggcggt ttggttcagc ccggtggatc actgcgactc 60

agttgcgcag ctagcggctt caccttttct gattactaca tgacatgggt acgacaggcg 120

ccaggcaagg gtttggaatg ggtagcattc atacgcaatc aggcacgcgg gtacacttca 180

gaccacaatc cctcagtaaa aggaagattt accatctcaa gagacaatgc caaaaattca 240

ctctacctgc aaatgaactc acttcgcgcc gaggataccg ccgtgtatta ctgtgccaga 300

gacagaccat cttattacgt gctggactat tggggacagg gcactacagt caccgtcagc 360

tct 363

<210> 292

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 292

gacatccaga tgacccagtc cccctcttct ctgtctgcct ctgtgggcga cagagtgacc 60

atcacctgca caagctcaca gtcactgttt aatgtccgca gcggcaaaaa ctatcttgcg 120

tggtatcagc agaagcctgg caaggctccc aagctgctga tctactgggc cagtgaccga 180

gaatccggcg tgccttccag attctccggc tctggctctg gcaccgattt caccctgacc 240

atctcctccc tccagcctga ggatttcgcc acctactact gcaaacagtc ttacgacctt 300

ttcacttttg gcggcggaac aaaggtggag atcaag 336

<210> 293

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic nucleotide sequence

<400> 293

gaagtgcagc ttgttgaatc tggcggcggt ttggttcagc ccggtggatc actgcgactc 60

agttgcgcag ctagcggctt caccttttct gattactaca tgacatgggt acgacaggcg 120

ccaggcaagg gtttggaatg ggtagcattc atacgcaatc aggcacgcgg gtacacttca 180

gaccacaatc cctcagtaaa aggaagattt accatctcaa gagacaatgc caaaaattca 240

ctctacctgc aaatgaactc acttcgcgcc gaggataccg ccgtgtatta ctgtgccaga 300

gacagaccat cttattacgt gctggactat tggggacagg gcactacagt caccgtcagc 360

tct 363

<210> 294

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 294

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Arg

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Lys Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Leu Ser Asp Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 295

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 295

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly His Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 296

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 296

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Pro Leu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 297

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 297

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 298

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 298

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Pro Leu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 299

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 299

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 300

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 300

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Arg Trp Asn Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 301

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 301

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 302

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 302

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Arg Gly Phe Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 303

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 303

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 304

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 304

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Thr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 305

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 305

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 306

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 306

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Pro

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Thr Gly Trp Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Thr Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 307

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 307

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Lys His Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 308

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 308

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Arg Gly Trp Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 309

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 309

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Arg His Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 310

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 310

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Thr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Arg Gly Phe Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Gln Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 311

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 311

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asn Trp Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 312

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 312

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Thr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Thr Lys Tyr Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Thr Ile Phe Thr Arg Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 313

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 313

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asn Trp Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 314

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 314

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Thr Lys Tyr Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Phe Thr Asn Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 315

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 315

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Ser Ile Tyr

20 25 30

Gly Ser Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 316

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 316

Met Lys Thr Leu Leu Leu Asp Leu Ala Leu Trp Ser Leu Leu Phe Gln

1 5 10 15

Pro Gly Trp Leu

20

<210> 317

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 317

Leu Asp Leu Ala Leu Trp Ser Leu Leu Phe Gln Pro Gly Trp Leu Ser

1 5 10 15

Phe Ser Ser Gln

20

<210> 318

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 318

Trp Ser Leu Leu Phe Gln Pro Gly Trp Leu Ser Phe Ser Ser Gln Val

1 5 10 15

Ser Gln Asn Cys

20

<210> 319

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 319

Gln Pro Gly Trp Leu Ser Phe Ser Ser Gln Val Ser Gln Asn Cys His

1 5 10 15

Asn Gly Ser Tyr

20

<210> 320

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 320

Ser Phe Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu

1 5 10 15

Ile Ser Val Leu

20

<210> 321

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 321

Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser Val Leu Met

1 5 10 15

Met Gly Asn Ser

20

<210> 322

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 322

His Asn Gly Ser Tyr Glu Ile Ser Val Leu Met Gly Asn Ser Ala

1 5 10 15

Phe Ala Glu Pro

20

<210> 323

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 323

Glu Ile Ser Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu

1 5 10 15

Lys Asn Leu Glu

20

<210> 324

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 324

Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu Glu Asp

1 5 10 15

Ala Val Asn Glu

20

<210> 325

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 325

Ala Phe Ala Glu Pro Leu Lys Asn Leu Glu Asp Ala Val Asn Glu Gly

1 5 10 15

Leu Glu Ile Val

20

<210> 326

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 326

Leu Lys Asn Leu Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg

1 5 10 15

Gly Arg Leu Gln

20

<210> 327

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 327

Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn

1 5 10 15

Ala Gly Leu Asn

20

<210> 328

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 328

Gly Leu Glu Ile Val Arg Gly Arg Leu Gln Asn Ala Gly Leu Asn Val

1 5 10 15

Thr Val Asn Ala

20

<210> 329

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 329

Arg Gly Arg Leu Gln Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr

1 5 10 15

Phe Met Tyr Ser

20

<210> 330

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 330

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

1 5 10 15

Gly Leu Ile His

20

<210> 331

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 331

Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp Gly Leu Ile His Asn

1 5 10 15

Ser Gly Asp Cys

20

<210> 332

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 332

Thr Phe Met Tyr Ser Asp Gly Leu Ile His Asn Ser Gly Asp Cys Arg

1 5 10 15

Ser Ser Thr Cys

20

<210> 333

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 333

Asp Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu

1 5 10 15

Gly Leu Asp Leu

20

<210> 334

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 334

Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu Asp Leu Leu

1 5 10 15

Arg Lys Ile Ser

20

<210> 335

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 335

Arg Ser Ser Thr Cys Glu Gly Leu Asp Leu Leu Arg Lys Ile Ser Asn

1 5 10 15

Ala Gln Arg Met

20

<210> 336

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 336

Glu Gly Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly

1 5 10 15

Cys Val Leu Ile

20

<210> 337

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 337

Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val Leu Ile Gly

1 5 10 15

Pro Ser Cys Thr

20

<210> 338

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 338

Asn Ala Gln Arg Met Gly Cys Val Leu Ile Gly Pro Ser Cys Thr Tyr

1 5 10 15

Ser Thr Phe Gln

20

<210> 339

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 339

Gly Cys Val Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met

1 5 10 15

Tyr Leu Asp Thr

20

<210> 340

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 340

Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr Glu

1 5 10 15

Leu Ser Tyr Pro

20

<210> 341

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 341

Tyr Ser Thr Phe Gln Met Tyr Leu Asp Thr Glu Leu Ser Tyr Pro Met

1 5 10 15

Ile Ser Ala Gly

20

<210> 342

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 342

Met Tyr Leu Asp Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser

1 5 10 15

Phe Gly Leu Ser

20

<210> 343

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 343

Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys

1 5 10 15

Asp Tyr Lys Glu

20

<210> 344

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 344

Met Ile Ser Ala Gly Ser Phe Gly Leu Ser Cys Asp Tyr Lys Glu Thr

1 5 10 15

Leu Thr Arg Leu

20

<210> 345

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 345

Ser Phe Gly Leu Ser Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met

1 5 10 15

Ser Pro Ala Arg

20

<210> 346

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 346

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

1 5 10 15

Leu Met Tyr Phe

20

<210> 347

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 347

Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys Leu Met Tyr Phe Leu

1 5 10 15

Val Asn Phe Trp

20

<210> 348

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 348

Met Ser Pro Ala Arg Lys Leu Met Tyr Phe Leu Val Asn Phe Trp Lys

1 5 10 15

Thr Asn Asp Leu

20

<210> 349

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 349

Lys Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro

1 5 10 15

Phe Lys Thr Tyr

20

<210> 350

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 350

Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe Lys Thr Tyr Ser

1 5 10 15

Trp Ser Thr Ser

20

<210> 351

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 351

Lys Thr Asn Asp Leu Pro Phe Lys Thr Tyr Ser Trp Ser Thr Ser Tyr

1 5 10 15

Val Tyr Lys Asn

20

<210> 352

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 352

Pro Phe Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly

1 5 10 15

Thr Glu Thr Glu

20

<210> 353

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 353

Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu Thr Glu Asp

1 5 10 15

Cys Phe Trp Tyr

20

<210> 354

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 354

Tyr Val Tyr Lys Asn Gly Thr Glu Thr Glu Asp Cys Phe Trp Tyr Leu

1 5 10 15

Asn Ala Leu Glu

20

<210> 355

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 355

Gly Thr Glu Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala

1 5 10 15

Ser Val Ser Tyr

20

<210> 356

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 356

Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr Phe

1 5 10 15

Ser His Glu Leu

20

<210> 357

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 357

Leu Asn Ala Leu Glu Ala Ser Val Ser Tyr Phe Ser His Glu Leu Gly

1 5 10 15

Phe Lys Val Val

20

<210> 358

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 358

Ala Ser Val Ser Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu

1 5 10 15

Arg Gln Asp Lys

20

<210> 359

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 359

Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys Glu

1 5 10 15

Phe Gln Asp Ile

20

<210> 360

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 360

Gly Phe Lys Val Val Leu Arg Gln Asp Lys Glu Phe Gln Asp Ile Leu

1 5 10 15

Met Asp His Asn

20

<210> 361

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 361

Leu Arg Gln Asp Lys Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg

1 5 10 15

Lys Ser Asn Val

20

<210> 362

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 362

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

1 5 10 15

Ile Met Cys Gly

20

<210> 363

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 363

Leu Met Asp His Asn Arg Lys Ser Asn Val Ile Ile Met Cys Gly Gly

1 5 10 15

Pro Glu Phe Leu

20

<210> 364

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 364

Arg Lys Ser Asn Val Ile Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr

1 5 10 15

Lys Leu Lys Gly

20

<210> 365

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 365

Ile Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp

1 5 10 15

Arg Ala Val Ala

20

<210> 366

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 366

Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg Ala Val Ala Glu

1 5 10 15

Asp Ile Val Ile

20

<210> 367

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 367

Tyr Lys Leu Lys Gly Asp Arg Ala Val Ala Glu Asp Ile Val Ile Ile

1 5 10 15

Leu Val Asp Leu

20

<210> 368

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 368

Asp Arg Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe

1 5 10 15

Asn Asp Gln Tyr

20

<210> 369

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 369

Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp Gln Tyr Leu

1 5 10 15

Glu Asp Asn Val

20

<210> 370

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 370

Ile Leu Val Asp Leu Phe Asn Asp Gln Tyr Leu Glu Asp Asn Val Thr

1 5 10 15

Ala Pro Asp Tyr

20

<210> 371

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 371

Phe Asn Asp Gln Tyr Leu Glu Asp Asn Val Thr Ala Pro Asp Tyr Met

1 5 10 15

Lys Asn Val Leu

20

<210> 372

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 372

Leu Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val Leu Val

1 5 10 15

Leu Thr Leu Ser

20

<210> 373

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 373

Thr Ala Pro Asp Tyr Met Lys Asn Val Leu Val Leu Thr Leu Ser Pro

1 5 10 15

Gly Asn Ser Leu

20

<210> 374

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 374

Met Lys Asn Val Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu

1 5 10 15

Asn Ser Ser Phe

20

<210> 375

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 375

Met Lys Asn Val Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu

1 5 10 15

Asn Ser Ser Phe

20

<210> 376

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 376

Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

1 5 10 15

Arg Asn Leu Ser

20

<210> 377

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 377

Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser Arg Asn Leu Ser Pro

1 5 10 15

Thr Lys Arg Asp

20

<210> 378

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 378

Leu Asn Ser Ser Phe Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe

1 5 10 15

Ala Leu Ala Tyr

20

<210> 379

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 379

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

1 5 10 15

Asn Gly Ile Leu

20

<210> 380

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 380

Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn Gly Ile Leu Leu

1 5 10 15

Phe Gly His Met

20

<210> 381

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 381

Phe Ala Leu Ala Tyr Leu Asn Gly Ile Leu Leu Phe Gly His Met Leu

1 5 10 15

Lys Ile Phe Leu

20

<210> 382

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 382

Leu Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu

1 5 10 15

Asn Gly Glu Asn

20

<210> 383

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 383

Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn Gly Glu Asn Ile

1 5 10 15

Thr Thr Pro Lys

20

<210> 384

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 384

Leu Lys Ile Phe Leu Glu Asn Gly Glu Asn Ile Thr Thr Pro Lys Phe

1 5 10 15

Ala His Ala Phe

20

<210> 385

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 385

Glu Asn Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg

1 5 10 15

Asn Leu Thr Phe

20

<210> 386

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 386

Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr Phe Glu

1 5 10 15

Gly Tyr Asp Gly

20

<210> 387

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 387

Phe Ala His Ala Phe Arg Asn Leu Thr Phe Glu Gly Tyr Asp Gly Pro

1 5 10 15

Val Thr Leu Asp

20

<210> 388

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 388

Arg Asn Leu Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp

1 5 10 15

Trp Gly Asp Val

20

<210> 389

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 389

Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val Asp

1 5 10 15

Ser Thr Met Val

20

<210> 390

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 390

Pro Val Thr Leu Asp Asp Trp Gly Asp Val Asp Ser Thr Met Val Leu

1 5 10 15

Leu Tyr Thr Ser

20

<210> 391

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 391

Asp Trp Gly Asp Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val

1 5 10 15

Asp Thr Lys Lys

20

<210> 392

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 392

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

1 5 10 15

Lys Val Leu Leu

20

<210> 393

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 393

Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr Lys Val Leu Leu Thr

1 5 10 15

Tyr Asp Thr His

20

<210> 394

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 394

Val Asp Thr Lys Lys Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val

1 5 10 15

Asn Lys Thr Tyr

20

<210> 395

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 395

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

1 5 10 15

Val Asp Met Ser

20

<210> 396

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 396

Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro Val Asp Met Ser Pro

1 5 10 15

Thr Phe Thr Trp

20

<210> 397

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 397

Val Asn Lys Thr Tyr Pro Val Asp Met Ser Pro Thr Phe Thr Trp Lys

1 5 10 15

Asn Ser Lys Leu

20

<210> 398

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 398

Pro Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro

1 5 10 15

Asn Asp Ile Thr

20

<210> 399

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 399

Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp Ile Thr Gly

1 5 10 15

Arg Gly Pro Gln

20

<210> 400

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 400

Lys Asn Ser Lys Leu Pro Asn Asp Ile Thr Gly Arg Gly Pro Gln Ile

1 5 10 15

Leu Met Ile Ala

20

<210> 401

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 401

Pro Asn Asp Ile Thr Gly Arg Gly Pro Gln Ile Leu Met Ile Ala Val

1 5 10 15

Phe Thr Leu Thr

20

<210> 402

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 402

Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val Leu Ile Gly

1 5 10 15

Pro Ser Cys Thr

20

<210> 403

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 403

Asn Ala Gln Arg Met Gly Cys Val Leu Ile Gly Pro Ser Cys Thr Tyr

1 5 10 15

Ser Thr Phe Gln

20

<210> 404

<211> 20

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 404

Gly Cys Val Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met

1 5 10 15

Tyr Leu Asp Thr

20

<210> 405

<211> 25

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 405

Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly Leu Asp Leu Leu

1 5 10 15

Arg Lys Ile Ser Asn Ala Gln Arg Met

20 25

<210> 406

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 406

Arg Ser Ser Thr Cys Glu Gly Leu Asp Leu Leu Arg Lys Ile Ser

1 5 10 15

<210> 407

<211> 254

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 407

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Thr Ser Ser Gln Ser Leu Phe Asn Val

20 25 30

Arg Ser Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Lys Gln

85 90 95

Ser Tyr Asp Leu Phe Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser

115 120 125

Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala

130 135 140

Ala Ser Gly Phe Thr Phe Ser Ser Tyr Trp Met His Trp Val Arg Gln

145 150 155 160

Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly Glu Ile Lys Pro Ser Asn

165 170 175

Glu Leu Thr Asn Val His Glu Lys Phe Lys Asp Arg Phe Thr Ile Ser

180 185 190

Val Asp Lys Ala Lys Asn Ser Ala Tyr Leu Gln Met Asn Ser Leu Arg

195 200 205

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg Thr Ile Thr Thr Thr

210 215 220

Glu Gly Tyr Trp Phe Phe Asp Val Trp Gly Gln Gly Thr Leu Val Thr

225 230 235 240

Val Ser Ser Gly Gly Cys Gly Gly His His His His His

245 250

<210> 408

<211> 252

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 408

Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Ser Ser Leu Leu Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Lys Leu Ala Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg

85 90 95

Lys Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly

100 105 110

Gly Ser Gly Gly Gly Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly

115 120 125

Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

130 135 140

Phe Thr Phe Ser Asp Tyr Tyr Met Thr Trp Val Arg Gln Ala Pro Gly

145 150 155 160

Lys Gly Leu Glu Trp Val Ala Phe Ile Arg Asn Gln Ala Arg Gly Tyr

165 170 175

Thr Ser Asp His Asn Pro Ser Val Lys Gly Arg Phe Thr Ile Ser Arg

180 185 190

Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala

195 200 205

Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Arg Pro Ser Tyr Tyr

210 215 220

Val Leu Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly

225 230 235 240

Gly Cys Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys

245 250

<210> 409

<211> 257

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 409

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Glu Leu Thr Asn Val His Glu Lys Phe

50 55 60

Lys Asp Arg Phe Thr Ile Ser Val Asp Lys Ala Lys Asn Ser Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Asp Ile Gln Leu Thr Gln

130 135 140

Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr

145 150 155 160

Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr Gly Ser Ser Leu Leu Gln

165 170 175

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala

180 185 190

Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly

195 200 205

Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp

210 215 220

Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Arg Lys Ala Tyr Thr Phe Gly

225 230 235 240

Gln Gly Thr Lys Leu Glu Ile Lys Thr Gly Ser Glu Asn Leu Tyr Phe

245 250 255

Gln

<210> 410

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 410

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 411

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 411

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Arg Gln Lys Cys His Asn Gly Thr Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Asp Asn Ser Ala Tyr Lys Glu Pro Leu Gln Asn Leu

65 70 75 80

Arg Asp Ala Val Glu Glu Gly Leu Asp Ile Val Arg Lys Arg Leu Arg

85 90 95

Glu Ala Glu Leu Asn Val Thr Val Asn Ala Thr Phe Ile Tyr Ser Asp

100 105 110

Gly Leu Ile His Lys Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Glu Ile Thr Arg Asp Arg Lys Met Gly Cys Val

130 135 140

Leu Met Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Asn Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Ile Leu Pro Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asp Phe Trp Lys Val Asn Asn Ala Pro Phe

195 200 205

Lys Thr Phe Ser Trp Asn Ser Ser Tyr Val Tyr Lys Asn Gly Ser Glu

210 215 220

Pro Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Gly Val Ser

225 230 235 240

Tyr Phe Ser Glu Val Leu Ser Phe Lys Asp Val Leu Arg Arg Ser Glu

245 250 255

Gln Phe Gln Glu Ile Leu Met Gly Arg Asn Arg Lys Ser Asn Val Ile

260 265 270

Val Met Cys Gly Thr Pro Glu Thr Phe Tyr Asn Val Lys Gly Asp Leu

275 280 285

Lys Val Ala Asp Asp Thr Val Val Ile Leu Val Asp Leu Phe Ser Asn

290 295 300

His Tyr Phe Glu Asp Asp Thr Arg Ala Pro Glu Tyr Met Asp Asn Val

305 310 315 320

Leu Val Leu Thr Leu Pro Pro Glu Lys Phe Ile Ala Asn Ala Ser Val

325 330 335

Ser Gly Arg Phe Pro Ser Glu Arg Ser Asp Phe Ser Leu Ala Tyr Leu

340 345 350

Glu Gly Thr Leu Leu Phe Gly His Met Leu Gln Thr Phe Leu Glu Asn

355 360 365

Gly Glu Ser Val Thr Thr Pro Lys Phe Ala Arg Ala Phe Arg Asn Leu

370 375 380

Thr Phe Gln Gly Leu Glu Gly Pro Val Thr Leu Asp Asp Ser Gly Asp

385 390 395 400

Ile Asp Asn Ile Met Cys Leu Leu Tyr Val Ser Leu Asp Thr Arg Lys

405 410 415

Tyr Lys Val Leu Met Ala Tyr Asp Thr His Lys Asn Gln Thr Ile Pro

420 425 430

Val Ala Thr Ser Pro Asn Phe Ile Trp Lys Asn His Arg Leu Pro Asn

435 440 445

Asp Val Pro Gly Leu Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 412

<211> 1028

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 412

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Ser Arg Cys Asn Asn Asn Asn Tyr Met Ile Asn Val Met Leu

50 55 60

Met Pro Asp Ser Asp Phe Pro Ser Thr Ser Glu Asn Leu Thr Ser Ala

65 70 75 80

Val Glu Glu Ala Leu Ser Thr Ile Gln Asn Glu Leu Glu Thr Glu Gly

85 90 95

Val Lys Val Thr Val Asn Ala Ser Phe His His Phe Arg Ser Ser Leu

100 105 110

Tyr Val Ser Gln Gly Cys Arg Thr Ser Thr Cys Glu Gly Val Glu Leu

115 120 125

Ile Lys Gln Ile Phe Glu Asn Gly Thr Leu Gly Cys Ala Val Ile Gly

130 135 140

Pro Ala Cys Thr Tyr Ala Thr Tyr Gln Met Val Ser Val Glu Thr Ile

145 150 155 160

Pro Ser Leu Pro Leu Ile Ser Val Gly Ser Phe Gly Leu Ser Cys Asp

165 170 175

Tyr Lys Glu Asn Leu Thr Arg Leu Leu Thr Pro Ala Arg Lys Val Asn

180 185 190

Asp Phe Phe Tyr Tyr Phe Trp Asn Glu Ile Gln Gln Pro Phe Lys Thr

195 200 205

Ser Thr Trp Glu Ser Val Tyr Leu Tyr Lys Lys Thr Asp Asn Ser Glu

210 215 220

Gln Cys Leu Trp Tyr Met Asn Ala Leu Asp Ala Gly Val Thr Gln Phe

225 230 235 240

Ser Glu Lys Leu Lys Phe Lys Asp Ile Val Arg Thr Gln Asp Gln Phe

245 250 255

Arg Arg Leu Val Lys Asn Pro Lys Arg Lys Ser Asn Val Ile Ile Met

260 265 270

Cys Gly Thr Pro Ala Asp Ile Arg Gln Asp Leu Gly Thr Glu Thr Val

275 280 285

Asp Lys Asp Ile Val Ile Ile Leu Ile Asp Leu Phe Lys Asn Thr Tyr

290 295 300

Phe Arg Asn Thr Thr Ser Ala Arg Tyr Met Gln Asn Val Leu Val Leu

305 310 315 320

Thr Leu Pro Pro Ala Asn Asn Asn Phe Ser Thr Arg Thr Thr Asp Thr

325 330 335

Ser Leu Leu Glu Asp Asp Phe Val Ile Gly Tyr Tyr Asn Ala Val Leu

340 345 350

Leu Phe Gly His Ile Leu Lys Lys Phe Ile Phe Ser Gln Ser Pro Val

355 360 365

Leu Pro Thr Ser Phe Ile Asn Glu Phe Arg Asn Ile Thr Phe Glu Gly

370 375 380

Ala Gln Gly Pro Val Thr Leu Asp Glu Phe Gly Asp Ile Asp Asn Asn

385 390 395 400

Leu Thr Leu Leu Tyr Thr Thr Gln Ser Ala Ser Asp Pro Gln Tyr Arg

405 410 415

Val Leu Met Tyr Phe Asn Thr Gln Glu Asn Asp Thr Tyr Val Val Ser

420 425 430

Thr Ser Pro Asp Phe Ile Trp Lys Ser His Arg Leu Pro Ser Asp Ile

435 440 445

Pro Ser Thr Gly Pro His Arg Thr Ser Asp Gln Ala Ser Gly Ala His

450 455 460

His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala

465 470 475 480

Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser Thr Pro

485 490 495

Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His Ser Thr

500 505 510

Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser His Ser

515 520 525

Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr Ser His

530 535 540

Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser Thr Ser

545 550 555 560

Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu Thr Ser

565 570 575

Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr Glu Thr

580 585 590

Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val Thr Glu

595 600 605

Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr Ser Ser

610 615 620

Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile Met Ser

625 630 635 640

Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val Tyr Thr

645 650 655

Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser Leu Ile

660 665 670

Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr Ser Ser

675 680 685

Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr Thr Pro

690 695 700

Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser Ser Thr

705 710 715 720

Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser Ser Val

725 730 735

Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr Pro Ser

740 745 750

Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser Thr Pro

755 760 765

Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His Ile Thr

770 775 780

Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser His Ser

785 790 795 800

Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr Ser Glu

805 810 815

Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val Ser Thr

820 825 830

Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr Ala Val

835 840 845

Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile Pro Thr

850 855 860

Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser Thr Ser

865 870 875 880

Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile Ser Thr

885 890 895

Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile Gln Ser

900 905 910

Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met Ser Thr

915 920 925

Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser Ser Phe

930 935 940

Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr Pro Pro

945 950 955 960

Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro Thr Thr

965 970 975

Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His Lys Leu

980 985 990

Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala Ala Asp

995 1000 1005

Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro Leu Val

1010 1015 1020

Leu Leu Ser Leu Leu

1025

<210> 413

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 413

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Asn

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Phe Asp

100 105 110

Gly Leu Leu His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Val Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 414

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 414

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asp Ser Asp Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Gly Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 415

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 415

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Val Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Asn Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 416

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 416

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Thr Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Ser Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 417

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 417

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Arg Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Ala Phe Leu Tyr Lys Leu Lys Gly Thr Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Arg Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 418

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 418

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Glu Phe Arg Asn Ile

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 419

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 419

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Thr Ser Thr Cys Glu Gly

115 120 125

Val Glu Leu Ile Lys Gln Ile Phe Glu Asn Gly Thr Leu Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 420

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 420

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Asn Glu Ile Gln Gln Pro Phe

195 200 205

Lys Thr Ser Thr Trp Glu Ser Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 421

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 421

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Lys Thr Asp Asn

210 215 220

Ser Glu Gln Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 422

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 422

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Lys Phe Lys Asp Ile Val Arg Thr Gln Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 423

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 423

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Asp

245 250 255

Gln Phe Arg Arg Leu Val Lys Asn Pro Lys Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 424

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 424

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Thr Glu

275 280 285

Thr Val Asp Lys Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 425

<211> 1029

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 425

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Arg Asn Thr Thr Ser Ala Arg Tyr Met Gln Asn Val Leu

305 310 315 320

Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe Ser

325 330 335

Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu Asn

340 345 350

Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn Gly

355 360 365

Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu Thr

370 375 380

Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp Val

385 390 395 400

Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys Tyr

405 410 415

Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro Val

420 425 430

Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn Asp

435 440 445

Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly Ala

450 455 460

His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp Tyr

465 470 475 480

Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser Thr

485 490 495

Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His Ser

500 505 510

Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser His

515 520 525

Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr Ser

530 535 540

His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser Thr

545 550 555 560

Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu Thr

565 570 575

Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr Glu

580 585 590

Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val Thr

595 600 605

Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr Ser

610 615 620

Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile Met

625 630 635 640

Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val Tyr

645 650 655

Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser Leu

660 665 670

Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr Ser

675 680 685

Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr Thr

690 695 700

Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser Ser

705 710 715 720

Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser Ser

725 730 735

Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr Pro

740 745 750

Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser Thr

755 760 765

Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His Ile

770 775 780

Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser His

785 790 795 800

Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr Ser

805 810 815

Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val Ser

820 825 830

Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr Ala

835 840 845

Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile Pro

850 855 860

Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser Thr

865 870 875 880

Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile Ser

885 890 895

Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile Gln

900 905 910

Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met Ser

915 920 925

Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser Ser

930 935 940

Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr Pro

945 950 955 960

Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro Thr

965 970 975

Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His Lys

980 985 990

Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala Ala

995 1000 1005

Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro Leu

1010 1015 1020

Val Leu Leu Ser Leu Leu

1025

<210> 426

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 426

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Ala Ser Asp Pro Gln

405 410 415

Tyr Arg Val Leu Met Tyr Phe Asn Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 427

<211> 1028

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 427

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Ser Arg Cys Asn Asn Asn Asn Tyr Met Ile Asn Val Met Leu

50 55 60

Met Pro Asp Ser Asp Phe Pro Ser Thr Ser Glu Asn Leu Thr Ser Ala

65 70 75 80

Val Glu Glu Ala Leu Ser Thr Ile Gln Asn Glu Leu Glu Thr Glu Gly

85 90 95

Val Lys Val Thr Val Asn Ala Ser Phe His His Phe Arg Ser Ser Leu

100 105 110

Tyr Val Ser Gln Gly Cys Arg Ser Ser Thr Cys Glu Gly Leu Asp Leu

115 120 125

Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Ala Val Ile Gly

130 135 140

Pro Ala Cys Thr Tyr Ala Thr Tyr Gln Met Val Ser Val Glu Thr Ile

145 150 155 160

Pro Ser Leu Pro Leu Ile Ser Val Gly Ser Phe Gly Leu Ser Cys Asp

165 170 175

Tyr Lys Glu Asn Leu Thr Arg Leu Leu Thr Pro Ala Arg Lys Val Asn

180 185 190

Asp Phe Phe Tyr Tyr Phe Trp Asn Glu Ile Gln Gln Pro Phe Lys Thr

195 200 205

Ser Thr Trp Glu Ser Val Tyr Leu Tyr Lys Lys Thr Asp Asn Ser Glu

210 215 220

Gln Cys Leu Trp Tyr Met Asn Ala Leu Asp Ala Gly Val Thr Gln Phe

225 230 235 240

Ser Glu Lys Leu Lys Phe Lys Asp Ile Val Arg Thr Gln Asp Gln Phe

245 250 255

Arg Arg Leu Val Lys Asn Pro Lys Arg Lys Ser Asn Val Ile Ile Met

260 265 270

Cys Gly Thr Pro Ala Asp Ile Arg Gln Asp Leu Gly Thr Glu Thr Val

275 280 285

Asp Lys Asp Ile Val Ile Ile Leu Ile Asp Leu Phe Lys Asn Thr Tyr

290 295 300

Phe Arg Asn Thr Thr Ser Ala Arg Tyr Met Gln Asn Val Leu Val Leu

305 310 315 320

Thr Leu Pro Pro Ala Asn Asn Asn Phe Ser Thr Arg Thr Thr Asp Thr

325 330 335

Ser Leu Leu Glu Asp Asp Phe Val Ile Gly Tyr Tyr Asn Ala Val Leu

340 345 350

Leu Phe Gly His Ile Leu Lys Lys Phe Ile Phe Ser Gln Ser Pro Val

355 360 365

Leu Pro Thr Ser Phe Ile Asn Glu Phe Arg Asn Ile Thr Phe Glu Gly

370 375 380

Ala Gln Gly Pro Val Thr Leu Asp Glu Phe Gly Asp Ile Asp Asn Asn

385 390 395 400

Leu Thr Leu Leu Tyr Thr Thr Gln Ser Ala Ser Asp Pro Gln Tyr Arg

405 410 415

Val Leu Met Tyr Phe Asn Thr Gln Glu Asn Asp Thr Tyr Val Val Ser

420 425 430

Thr Ser Pro Asp Phe Ile Trp Lys Ser His Arg Leu Pro Ser Asp Ile

435 440 445

Pro Ser Thr Gly Pro His Arg Thr Ser Asp Gln Ala Ser Gly Ala His

450 455 460

His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala

465 470 475 480

Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser Thr Pro

485 490 495

Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His Ser Thr

500 505 510

Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser His Ser

515 520 525

Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr Ser His

530 535 540

Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser Thr Ser

545 550 555 560

Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu Thr Ser

565 570 575

Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr Glu Thr

580 585 590

Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val Thr Glu

595 600 605

Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr Ser Ser

610 615 620

Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile Met Ser

625 630 635 640

Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val Tyr Thr

645 650 655

Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser Leu Ile

660 665 670

Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr Ser Ser

675 680 685

Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr Thr Pro

690 695 700

Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser Ser Thr

705 710 715 720

Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser Ser Val

725 730 735

Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr Pro Ser

740 745 750

Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser Thr Pro

755 760 765

Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His Ile Thr

770 775 780

Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser His Ser

785 790 795 800

Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr Ser Glu

805 810 815

Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val Ser Thr

820 825 830

Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr Ala Val

835 840 845

Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile Pro Thr

850 855 860

Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser Thr Ser

865 870 875 880

Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile Ser Thr

885 890 895

Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile Gln Ser

900 905 910

Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met Ser Thr

915 920 925

Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser Ser Phe

930 935 940

Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr Pro Pro

945 950 955 960

Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro Thr Thr

965 970 975

Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His Lys Leu

980 985 990

Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala Ala Asp

995 1000 1005

Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro Leu Val

1010 1015 1020

Leu Leu Ser Leu Leu

1025

<210> 428

<211> 1028

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 428

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Ser Arg Cys Asn Asn Asn Asn Tyr Met Ile Asn Val Met Leu

50 55 60

Met Pro Asp Ser Asp Phe Pro Ser Thr Ser Glu Asn Leu Thr Ser Ala

65 70 75 80

Val Glu Glu Ala Leu Ser Thr Ile Gln Asn Glu Leu Glu Thr Glu Gly

85 90 95

Val Lys Val Thr Val Asn Ala Ser Phe His His Phe Arg Ser Ser Leu

100 105 110

Tyr Val Ser Gln Gly Cys Arg Ser Ser Thr Cys Glu Gly Leu Asp Leu

115 120 125

Leu Arg Lys Ile Ser Glu Asn Gly Thr Leu Gly Cys Ala Val Ile Gly

130 135 140

Pro Ala Cys Thr Tyr Ala Thr Tyr Gln Met Val Ser Val Glu Thr Ile

145 150 155 160

Pro Ser Leu Pro Leu Ile Ser Val Gly Ser Phe Gly Leu Ser Cys Asp

165 170 175

Tyr Lys Glu Asn Leu Thr Arg Leu Leu Thr Pro Ala Arg Lys Val Asn

180 185 190

Asp Phe Phe Tyr Tyr Phe Trp Asn Glu Ile Gln Gln Pro Phe Lys Thr

195 200 205

Ser Thr Trp Glu Ser Val Tyr Leu Tyr Lys Lys Thr Asp Asn Ser Glu

210 215 220

Gln Cys Leu Trp Tyr Met Asn Ala Leu Asp Ala Gly Val Thr Gln Phe

225 230 235 240

Ser Glu Lys Leu Lys Phe Lys Asp Ile Val Arg Thr Gln Asp Gln Phe

245 250 255

Arg Arg Leu Val Lys Asn Pro Lys Arg Lys Ser Asn Val Ile Ile Met

260 265 270

Cys Gly Thr Pro Ala Asp Ile Arg Gln Asp Leu Gly Thr Glu Thr Val

275 280 285

Asp Lys Asp Ile Val Ile Ile Leu Ile Asp Leu Phe Lys Asn Thr Tyr

290 295 300

Phe Arg Asn Thr Thr Ser Ala Arg Tyr Met Gln Asn Val Leu Val Leu

305 310 315 320

Thr Leu Pro Pro Ala Asn Asn Asn Phe Ser Thr Arg Thr Thr Asp Thr

325 330 335

Ser Leu Leu Glu Asp Asp Phe Val Ile Gly Tyr Tyr Asn Ala Val Leu

340 345 350

Leu Phe Gly His Ile Leu Lys Lys Phe Ile Phe Ser Gln Ser Pro Val

355 360 365

Leu Pro Thr Ser Phe Ile Asn Glu Phe Arg Asn Ile Thr Phe Glu Gly

370 375 380

Ala Gln Gly Pro Val Thr Leu Asp Glu Phe Gly Asp Ile Asp Asn Asn

385 390 395 400

Leu Thr Leu Leu Tyr Thr Thr Gln Ser Ala Ser Asp Pro Gln Tyr Arg

405 410 415

Val Leu Met Tyr Phe Asn Thr Gln Glu Asn Asp Thr Tyr Val Val Ser

420 425 430

Thr Ser Pro Asp Phe Ile Trp Lys Ser His Arg Leu Pro Ser Asp Ile

435 440 445

Pro Ser Thr Gly Pro His Arg Thr Ser Asp Gln Ala Ser Gly Ala His

450 455 460

His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp Tyr Ala

465 470 475 480

Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser Thr Pro

485 490 495

Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His Ser Thr

500 505 510

Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser His Ser

515 520 525

Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr Ser His

530 535 540

Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser Thr Ser

545 550 555 560

Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu Thr Ser

565 570 575

Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr Glu Thr

580 585 590

Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val Thr Glu

595 600 605

Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr Ser Ser

610 615 620

Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile Met Ser

625 630 635 640

Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val Tyr Thr

645 650 655

Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser Leu Ile

660 665 670

Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr Ser Ser

675 680 685

Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr Thr Pro

690 695 700

Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser Ser Thr

705 710 715 720

Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser Ser Val

725 730 735

Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr Pro Ser

740 745 750

Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser Thr Pro

755 760 765

Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His Ile Thr

770 775 780

Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser His Ser

785 790 795 800

Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr Ser Glu

805 810 815

Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val Ser Thr

820 825 830

Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr Ala Val

835 840 845

Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile Pro Thr

850 855 860

Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser Thr Ser

865 870 875 880

Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile Ser Thr

885 890 895

Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile Gln Ser

900 905 910

Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met Ser Thr

915 920 925

Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser Ser Phe

930 935 940

Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr Pro Pro

945 950 955 960

Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro Thr Thr

965 970 975

Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His Lys Leu

980 985 990

Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala Ala Asp

995 1000 1005

Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro Leu Val

1010 1015 1020

Leu Leu Ser Leu Leu

1025

<210> 429

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 429

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Asn

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 430

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 430

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Phe Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 431

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 431

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Leu His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 432

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 432

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Thr Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 433

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 433

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Val Asp Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 434

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 434

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Glu Leu Leu Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 435

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 435

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Ile Arg Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 436

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 436

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Lys Lys Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 437

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 437

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Gln Ile Ser Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 438

<211> 1030

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 438

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Ser Gln Val Ser Gln Asn Cys His Asn Gly Ser Tyr Glu Ile Ser

50 55 60

Val Leu Met Met Gly Asn Ser Ala Phe Ala Glu Pro Leu Lys Asn Leu

65 70 75 80

Glu Asp Ala Val Asn Glu Gly Leu Glu Ile Val Arg Gly Arg Leu Gln

85 90 95

Asn Ala Gly Leu Asn Val Thr Val Asn Ala Thr Phe Met Tyr Ser Asp

100 105 110

Gly Leu Ile His Asn Ser Gly Asp Cys Arg Ser Ser Thr Cys Glu Gly

115 120 125

Leu Asp Leu Leu Arg Lys Ile Phe Asn Ala Gln Arg Met Gly Cys Val

130 135 140

Leu Ile Gly Pro Ser Cys Thr Tyr Ser Thr Phe Gln Met Tyr Leu Asp

145 150 155 160

Thr Glu Leu Ser Tyr Pro Met Ile Ser Ala Gly Ser Phe Gly Leu Ser

165 170 175

Cys Asp Tyr Lys Glu Thr Leu Thr Arg Leu Met Ser Pro Ala Arg Lys

180 185 190

Leu Met Tyr Phe Leu Val Asn Phe Trp Lys Thr Asn Asp Leu Pro Phe

195 200 205

Lys Thr Tyr Ser Trp Ser Thr Ser Tyr Val Tyr Lys Asn Gly Thr Glu

210 215 220

Thr Glu Asp Cys Phe Trp Tyr Leu Asn Ala Leu Glu Ala Ser Val Ser

225 230 235 240

Tyr Phe Ser His Glu Leu Gly Phe Lys Val Val Leu Arg Gln Asp Lys

245 250 255

Glu Phe Gln Asp Ile Leu Met Asp His Asn Arg Lys Ser Asn Val Ile

260 265 270

Ile Met Cys Gly Gly Pro Glu Phe Leu Tyr Lys Leu Lys Gly Asp Arg

275 280 285

Ala Val Ala Glu Asp Ile Val Ile Ile Leu Val Asp Leu Phe Asn Asp

290 295 300

Gln Tyr Phe Glu Asp Asn Val Thr Ala Pro Asp Tyr Met Lys Asn Val

305 310 315 320

Leu Val Leu Thr Leu Ser Pro Gly Asn Ser Leu Leu Asn Ser Ser Phe

325 330 335

Ser Arg Asn Leu Ser Pro Thr Lys Arg Asp Phe Ala Leu Ala Tyr Leu

340 345 350

Asn Gly Ile Leu Leu Phe Gly His Met Leu Lys Ile Phe Leu Glu Asn

355 360 365

Gly Glu Asn Ile Thr Thr Pro Lys Phe Ala His Ala Phe Arg Asn Leu

370 375 380

Thr Phe Glu Gly Tyr Asp Gly Pro Val Thr Leu Asp Asp Trp Gly Asp

385 390 395 400

Val Asp Ser Thr Met Val Leu Leu Tyr Thr Ser Val Asp Thr Lys Lys

405 410 415

Tyr Lys Val Leu Leu Thr Tyr Asp Thr His Val Asn Lys Thr Tyr Pro

420 425 430

Val Asp Met Ser Pro Thr Phe Thr Trp Lys Asn Ser Lys Leu Pro Asn

435 440 445

Asp Ile Thr Gly Arg Gly Pro Gln Arg Thr Ser Asp Gln Ala Ser Gly

450 455 460

Ala His His His His His Gly Ala Tyr Pro Tyr Asp Val Pro Asp

465 470 475 480

Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly Gly Thr Ser

485 490 495

Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr Pro Ser His

500 505 510

Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu Thr Pro Ser

515 520 525

His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser Glu Thr Thr

530 535 540

Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr Thr Glu Ser

545 550 555 560

Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr Ile Thr Glu

565 570 575

Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile Ser Ser Thr

580 585 590

Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser Ile Thr Val

595 600 605

Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser Thr Leu Thr

610 615 620

Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr Ser Ser Ile

625 630 635 640

Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu Thr Ser Val

645 650 655

Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr Ser Thr Ser

660 665 670

Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro Ser Phe Thr

675 680 685

Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala Ser Ser Tyr

690 695 700

Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn Thr Thr Ser

705 710 715 720

Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser Ser Ser Ser

725 730 735

Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser Tyr Ser Thr

740 745 750

Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro Leu His Ser

755 760 765

Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr Thr Ser His

770 775 780

Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu Thr Tyr Ser

785 790 795 800

His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr Glu Ser Thr

805 810 815

Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr Ser Thr Val

820 825 830

Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr Ser Glu Thr

835 840 845

Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser Thr Asp Ile

850 855 860

Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val Gly Thr Ser

865 870 875 880

Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro Thr Asp Ile

885 890 895

Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser Pro Ser Ile

900 905 910

Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser Pro Thr Met

915 920 925

Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr Pro Ile Ser

930 935 940

Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro Thr Gln Thr

945 950 955 960

Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser Pro Ala Pro

965 970 975

Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser Thr Leu His

980 985 990

Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr Ile Gly Ala

995 1000 1005

Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu Ala Pro

1010 1015 1020

Leu Val Leu Leu Ser Leu Leu

1025 1030

<210> 439

<211> 617

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 439

Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val

1 5 10 15

Leu Ala Gln Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp

20 25 30

Ser Thr Gly Gly Gly Ser Gly Gly Gly Ala Gln Ala Ala Gly Ala His

35 40 45

Ser Arg Thr Ser Asp Gln Ala Ser Gly Ala Gly Ala Tyr Pro Tyr Asp

50 55 60

Val Pro Asp Tyr Ala Gly His Gly Thr Ser Gly Gly Gly Gly Ser Gly

65 70 75 80

Gly Thr Ser Thr Pro Ser Tyr Thr Thr Ser Ile Ile Ser Thr Glu Thr

85 90 95

Pro Ser His Ser Thr Pro Ser Ser Ser Thr Ser Ile Thr Thr Thr Glu

100 105 110

Thr Pro Ser His Ser Thr Pro Ser Tyr Thr Ser Ser Val Ser Thr Ser

115 120 125

Glu Thr Thr Ser His Ser Thr Pro Ser Glu Thr Ser Ser Ser Arg Thr

130 135 140

Thr Glu Ser Thr Ser Tyr Ser Ser Pro Ser Ser Thr Ser Ser Asn Thr

145 150 155 160

Ile Thr Glu Thr Ser Ser His Ser Thr Pro Ser Thr Ala Thr Ser Ile

165 170 175

Ser Ser Thr Glu Thr Pro Ser Ser Ser Ile Pro Ser Val Ser Ser Ser

180 185 190

Ile Thr Val Thr Glu Ser Ser Ser His Ser Thr Pro Gly Ala Thr Ser

195 200 205

Thr Leu Thr Ser Ser Glu Thr Ser Thr Trp Ser Thr Pro Ser Ser Thr

210 215 220

Ser Ser Ile Met Ser Ser Ser Tyr Thr Ser Ala Asp Thr Pro Ser Glu

225 230 235 240

Thr Ser Val Tyr Thr Ser Ser Glu Thr Pro Ser Ser Ser Ser Pro Thr

245 250 255

Ser Thr Ser Leu Ile Ser Ser Ser Lys Ser Thr Ser Thr Ser Thr Pro

260 265 270

Ser Phe Thr Ser Ser Ile Thr Ser Thr Glu Thr Ser Ser Tyr Ser Ala

275 280 285

Ser Ser Tyr Thr Pro Ser Val Ser Ser Thr Ala Ser Ser Ser Lys Asn

290 295 300

Thr Thr Ser Ser Thr Ala Ser Ile Ser Ser Ala Glu Thr Val Ser Ser

305 310 315 320

Ser Ser Ser Ser Val Ser Ser Thr Ile Pro Ser Ser Gln Ser Thr Ser

325 330 335

Tyr Ser Thr Pro Ser Phe Ser Ser Ser Ala Thr Ser Ser Val Thr Pro

340 345 350

Leu His Ser Thr Pro Ser Leu Pro Ser Trp Val Thr Thr Ser Lys Thr

355 360 365

Thr Ser His Ile Thr Pro Gly Leu Thr Ser Ser Met Ser Ser Ser Glu

370 375 380

Thr Tyr Ser His Ser Thr Pro Gly Phe Thr Ser Ser Ile Thr Ser Thr

385 390 395 400

Glu Ser Thr Ser Glu Ser Thr Pro Ser Leu Ser Ser Ser Thr Ile Tyr

405 410 415

Ser Thr Val Ser Thr Ser Thr Thr Ala Ile Thr Ser His Phe Thr Thr

420 425 430

Ser Glu Thr Ala Val Thr Pro Thr Pro Val Thr Pro Ser Ser Leu Ser

435 440 445

Thr Asp Ile Pro Thr Thr Ser Leu Arg Thr Leu Thr Pro Ser Ser Val

450 455 460

Gly Thr Ser Thr Ser Leu Thr Thr Thr Thr Asp Phe Pro Ser Ile Pro

465 470 475 480

Thr Asp Ile Ser Thr Leu Pro Thr Arg Thr His Ile Ile Ser Ser Ser

485 490 495

Pro Ser Ile Gln Ser Thr Glu Thr Ser Ser Leu Val Gly Thr Thr Ser

500 505 510

Pro Thr Met Ser Thr Val Arg Met Thr Leu Arg Ile Thr Glu Asn Thr

515 520 525

Pro Ile Ser Ser Phe Ser Thr Ser Ile Val Val Ile Pro Glu Thr Pro

530 535 540

Thr Gln Thr Pro Pro Val Leu Thr Ser Ala Thr Gly Thr Gln Thr Ser

545 550 555 560

Pro Ala Pro Thr Thr Val Thr Phe Gly Ser Thr Asp Ser Ser Thr Ser

565 570 575

Thr Leu His Lys Leu Gln Gly Asp Ser Gly Glu Thr Ile Gly Lys Tyr

580 585 590

Ile Gly Ala Ala Asp Ser Leu Gly Gly Ser Val Leu Leu Leu Ala Leu

595 600 605

Ala Pro Leu Val Leu Leu Ser Leu Leu

610 615

<210> 440

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 440

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Gly Asp

100 105 110

Pro Val Ala Pro Thr Val Leu Ile Phe Pro Pro Ala Ala Asp Gln Val

115 120 125

Ala Thr Gly Thr Val Thr Ile Val Cys Val Ala Asn Lys Tyr Phe Pro

130 135 140

Asp Val Thr Val Thr Trp Glu Val Asp Gly Thr Thr Gln Thr Thr Gly

145 150 155 160

Ile Glu Asn Ser Lys Thr Pro Gln Asn Ser Ala Asp Cys Thr Tyr Asn

165 170 175

Leu Ser Ser Thr Leu Thr Leu Thr Ser Thr Gln Tyr Asn Ser His Lys

180 185 190

Glu Tyr Thr Cys Arg Val Thr Gln Gly Thr Thr Ser Val Val Gln Ser

195 200 205

Phe Asn Arg Gly Asp Cys

210

<210> 441

<211> 445

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 441

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Lys Ser Ala Thr Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ala Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Gln Pro Lys Ala Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Cys Cys Gly Asp Thr Pro Ser Ser Thr

130 135 140

Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Leu Pro Glu Pro Val Thr

145 150 155 160

Val Thr Trp Asn Ser Gly Thr Leu Thr Asn Gly Val Arg Thr Phe Pro

165 170 175

Ser Val Arg Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Ser

180 185 190

Val Thr Ser Ser Ser Gln Pro Val Thr Cys Asn Val Ala His Pro Ala

195 200 205

Thr Asn Thr Lys Val Asp Lys Thr Val Ala Pro Ser Thr Cys Ser Lys

210 215 220

Pro Thr Cys Pro Pro Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Ile

225 230 235 240

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu

245 250 255

Val Thr Cys Val Val Val Asp Val Ser Gln Asp Asp Pro Glu Val Gln

260 265 270

Phe Thr Trp Tyr Ile Asn Asn Glu Gln Val Arg Thr Ala Arg Pro Pro

275 280 285

Leu Arg Glu Gln Gln Phe Asn Ser Thr Ile Arg Val Val Ser Thr Leu

290 295 300

Pro Ile Ala His Gln Asp Trp Leu Arg Gly Lys Glu Phe Lys Cys Lys

305 310 315 320

Val His Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys

325 330 335

Ala Arg Gly Gln Pro Leu Glu Pro Lys Val Tyr Thr Met Gly Pro Pro

340 345 350

Arg Glu Glu Leu Ser Ser Arg Ser Val Ser Leu Thr Cys Met Ile Asn

355 360 365

Gly Phe Tyr Pro Ser Asp Ile Ser Val Glu Trp Glu Lys Asn Gly Lys

370 375 380

Ala Glu Asp Asn Tyr Lys Thr Thr Pro Ala Val Leu Asp Ser Asp Gly

385 390 395 400

Ser Tyr Phe Leu Tyr Ser Lys Leu Ser Val Pro Thr Ser Glu Trp Gln

405 410 415

Arg Gly Asp Val Phe Thr Cys Ser Val Met His Glu Ala Leu His Asn

420 425 430

His Tyr Thr Gln Lys Ser Ile Ser Arg Ser Pro Gly Lys

435 440 445

<210> 442

<211> 217

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 442

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr

100 105 110

Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu

115 120 125

Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro

130 135 140

Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn

145 150 155 160

Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His

180 185 190

Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile

195 200 205

Val Lys Ser Phe Asn Arg Asn Glu Cys

210 215

<210> 443

<211> 446

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 443

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Lys Ser Ala Thr Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ala Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro

115 120 125

Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met

130 135 140

Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val

180 185 190

Pro Ser Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His

195 200 205

Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys

210 215 220

Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe

225 230 235 240

Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro

245 250 255

Lys Val Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val

260 265 270

Gln Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr

275 280 285

Gln Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu

290 295 300

Leu Pro Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys

305 310 315 320

Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Thr Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro

340 345 350

Pro Lys Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile

355 360 365

Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly

370 375 380

Gln Pro Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp

385 390 395 400

Gly Ser Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp

405 410 415

Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His

420 425 430

Asn His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys

435 440 445

<210> 444

<211> 217

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 444

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Ser Leu Asn Ile His

65 70 75 80

Pro Val Glu Glu Asp Asp Ile Ala Met Tyr Phe Cys Gln Gln Thr Arg

85 90 95

Lys Val Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr

100 105 110

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

115 120 125

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

130 135 140

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

145 150 155 160

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

165 170 175

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

180 185 190

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

195 200 205

Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 445

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Synthetic peptide sequence

<400> 445

Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Lys Pro Ser Asn Gly Leu Thr Asn Tyr Ile Glu Lys Phe

50 55 60

Lys Asn Lys Ala Thr Leu Thr Val Asp Lys Ser Ala Thr Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ala Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Thr Ile Thr Thr Thr Glu Gly Tyr Trp Phe Phe Asp Val Trp

100 105 110

Gly Ala Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

225 230 235 240

Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly

450

<---

Claims (152)

1. An antibody that specifically binds to guanylate cyclase C (GUCY2c), where the antibody contains: a. a heavy chain variable region (VH) comprising a VH complementarity determining region one (VH CDR1), a VH complementarity determining region two (VH CDR2), and a VH complementarity determining region three (VH CDR3) of the VH sequence of SEQ ID NO:73; And b. a light chain variable region (VL) comprising a VL complementarity determining region one (VL CDR1), a VL complementarity determining region two (VL CDR2), and a VL complementarity determining region three (VL CDR3) of the VL sequence of SEQ ID NO:147. 2. Antibody according to claim 1, where: a. region VH contains (i) CDR1 VH containing the sequence of SEQ ID NO:74 or 259; (ii) CDR2 VH containing the sequence of SEQ ID NO:75 or 267; And (iii) CDR3 VH containing the sequence SEQ ID NO:29; And b. region VL contains (i) CDR1 VL containing the sequence of SEQ ID NO:148; (ii) CDR2 VL containing the sequence of SEQ ID NO:149; And (iii) CDR3 VL containing the sequence of SEQ ID NO:142. 3. Antibody according to claim 1, where: A. the VH region contains the sequence SEQ ID NO:73; And b. the VL region contains the sequence SEQ ID NO:147. 4. Antibody according to claim 1, where the VH region contains the sequence SEQ ID NO:73; And the VL region contains the sequence SEQ ID NO:147. 5. Antibody according to claim 1, where the VH region contains the sequence SEQ ID NO:73 or a variant thereof with one or more conservative amino acid substitutions at residues that are not in the CDR region; and/or the VL region contains the sequence SEQ ID NO:147 or a variant thereof with one or more conservative amino acid substitutions in amino acids that are not in the CDR region. 6. The antibody according to claim 1, where the antibody is selected from the group consisting of a Fab fragment, a Fab' fragment, an F(ab') ₂ fragment, an Fd fragment, an Fv fragment, a single chain Fv fragment (scFv), disulfide bond stabilized Fv fragment (dsFv), single domain antibody fragment (dAb), monoclonal antibody, chimeric antibody, polyclonal antibody, humanized antibody, human antibody and fragments thereof. 7. Antibody according to claim 1, containing human VH scaffold or humanized VH scaffold and a human VL scaffold or a humanized VL scaffold. 8. Antibody according to claim 7, where VH framework contains the sequence SEQ ID NO: 5, 6, 7, 8, 15, 16, 17, 18, 23, 24, 25, 30, 31, 32, 37, 38, 39, 40, 45, 46, 47, 49, 50, 51, 54, 55, 56, 58, 59, 61 or 63; and/or VL framework contains the sequence SEQ ID NO: 80, 81, 82, 83, 86, 87, 88, 89, 96, 97, 98, 99, 103, 110, 111, 116, 117, 118, 122, 123, 124, 126, 127, 128, 130, 131, 132, 133, 135, 139 or 155. 9. An isolated human monoclonal antibody according to any one of claims 1 to 8, which binds to an epitope on the extracellular domain of GUCY2c, where the epitope contains at least one amino acid residue selected from amino acid residues R73, S74, S75, T76, E78, G79, L80, L82, L83, R84 or I86 of the sequence SEQ ID NO:406. 10. A bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain, wherein: a. (i) the first polypeptide chain contains, in a direction from N-terminus to C-terminus: domain 1 containing the VL region of the anti-GUCY2c antibody (VL GUCY2c) and the VH region of the anti-CD3 antibody (VH CD3), and the first heterodimerization promoting domain; And (ii) the second polypeptide chain contains, in an N-terminal to C-terminal direction: domain 2 containing the VL region of the anti-CD3 antibody (VL CD3) and the VH region of the anti-GUCY2c antibody (VH GUCY2c), and a second heterodimerization promoting domain; b. (i) the first polypeptide chain contains, in a direction from N-terminus to C-terminus: domain 1 containing CD3 VL and GUCY2c VH, and a first heterodimerization promoting domain; And (ii) the second polypeptide chain contains, in an N-terminal to C-terminal direction: domain 2 containing GUCY2c VL and CD3 VH, and a second heterodimerization promoting domain; With. (i) the first polypeptide chain contains, in a direction from N-terminus to C-terminus: domain 1 containing VH GUCY2c and VL CD3, and a first heterodimerization promoting domain; And (ii) the second polypeptide chain contains, in a direction from the N-terminus to the C-terminus: domain 2 containing VH CD3 and VL GUCY2c, and a second heterodimerization promoting domain; or d. (i) the first polypeptide chain contains, in a direction from N-terminus to C-terminus: domain 1 containing CD3 VH and GUCY2c VL, and a first heterodimerization promoting domain; And (ii) the second polypeptide chain contains, in an N-terminal to C-terminal direction: domain 2 containing GUCY2c VH and CD3 VL, and a second heterodimerization promoting domain, and where VL GUCY2c and VH GUCY2c form a domain that specifically binds to GUCY2c; and VL CD3 and VH CD3 form a domain that specifically binds to CD3, and where VH GUCY2c contains: (i) CDR1 VH GUCY2c contains the sequence of SEQ ID NO:74 or 259; (ii) CDR2 VH GUCY2c contains the sequence SEQ ID NO:75 or 267; (iii) CDR3 VH GUCY2c contains the sequence SEQ ID NO:29; VL GUCY2c contains: (i) CDR1 VL GUCY2c contains the sequence SEQ ID NO:148; (ii) CDR2 VL GUCY2c contains the sequence SEQ ID NO:149; And (iii) CDR3 VL GUCY2c contains the sequence SEQ ID NO:142; VH CD3 contains (i) CDR1 VH CD3 contains the sequence of SEQ ID NO:2 or 268; (ii) CDR2 VH CD3 contains the sequence SEQ ID NO:10 or 270; And (iii) CDR3 VH CD3 contains the sequence SEQ ID NO:4; And VL CD3 contains (i) CDR1 VL CD3 contains the sequence SEQ ID NO:91; (ii) CDR2 VL CD3 contains the sequence SEQ ID NO:78; And (iii) CDR3 VL CD3 contains the sequence SEQ ID NO:79. 11. The bispecific antibody of claim 10, wherein each of the first heterodimerization promoting domain and the second heterodimerization promoting domain comprises an Fc region comprising a CH2 domain and a CH3 domain, wherein the amino acid sequence of each of the CH2 and/or CH3 domains contains at least one amino acid modification compared to the wild type Fc region, resulting in the formation of a protrusion or a recess. 12. The bispecific antibody according to claim 11, wherein both the first heterodimerization-promoting domain and the second heterodimerization-promoting domain are not protrusions or recesses; and/or wherein the first heterodimerization-promoting domain and the second heterodimerization-promoting domain form the Fc region of an IgG immunoglobulin. 13. Bispecific antibody according to claim 10, where VL GUCY2c and VH CD3 are connected by a glycine-serine linker; And VL CD3 and VH GUCY2c are connected by a glycine-serine linker. 14. Bispecific antibody according to claim 10, where domain 1 is covalently linked to the first heterodimerization promoting domain via a cysteine linker; And domain 2 is covalently linked to the second heterodimerization promoting domain via a cysteine linker; wherein the cysteine linker contains at least five amino acids. 15. Bispecific antibody according to claim 10, where: (i) the first polypeptide chain contains, in a direction from N-terminus to C-terminus: domain 1 containing the VL region of the GUCY2c antibody (VL GUCY2c) and the VH region of the anti-CD3 antibody (VH CD3), and a first heterodimerization promoting domain; And (ii) the second polypeptide chain contains, in an N-terminal to C-terminal direction: domain 2 containing the VL region of the anti-CD3 antibody (VL CD3) and the VH region of the GUCY2c antibody (VH GUCY2c), and a second heterodimerization promoting domain; and where VH GUCY2c contains (i) CDR1 VH GUCY2c containing the amino acid sequence of SEQ ID NO:259, (ii) CDR2 VH GUCY2c containing the amino acid sequence of SEQ ID NO:75, and (iii) CDR3 VH GUCY2c containing the amino acid sequence of SEQ ID NO:29; VL GUCY2c contains (i) CDR1 VL GUCY2c containing the amino acid sequence of SEQ ID NO:148, (ii) CDR2 VL GUCY2c containing the amino acid sequence of SEQ ID NO:149, and (iii) CDR3 VL GUCY2c containing the amino acid sequence of SEQ ID NO:142; VH CD3 contains (i) CDR1 VH CD3 containing the amino acid sequence of SEQ ID NO:268, (ii) CDR2 VH CD3 containing the amino acid sequence of SEQ ID NO:10, and (iii) CDR3 VH CD3 containing the amino acid sequence of SEQ ID NO:4; And VL CD3 contains (i) CDR1 VL CD3 containing the amino acid sequence of SEQ ID NO:91, (ii) CDR2 VL CD3 containing the amino acid sequence of SEQ ID NO:78, and (iii) CDR3 VL CD3 containing the amino acid sequence of SEQ ID NO:79. 16. Bispecific antibody according to claim 10, where (i) the first polypeptide chain contains, in a direction from N-terminus to C-terminus: domain 1 containing the VL region of the GUCY2c antibody (VL GUCY2c) and the VH region of the anti-CD3 antibody (VH CD3), and a first heterodimerization promoting domain; And (ii) the second polypeptide chain contains, in an N-terminal to C-terminal direction: domain 2 containing the VL region of the anti-CD3 antibody (VL CD3) and the VH region of the GUCY2c antibody (VH GUCY2c), and a second heterodimerization promoting domain; and where VH GUCY2c contains (i) CDR1 VH GUCY2c containing the amino acid sequence of SEQ ID NO:74, (ii) CDR2 VH GUCY2c containing the amino acid sequence of SEQ ID NO:75, and (iii) CDR3 VH GUCY2c containing the amino acid sequence of SEQ ID NO:29; VL GUCY2c contains (i) CDR1 VL GUCY2c containing the amino acid sequence of SEQ ID NO:148, (ii) CDR2 VL GUCY2c containing the amino acid sequence of SEQ ID NO:149, and (iii) CDR3 VL GUCY2c containing the amino acid sequence of SEQ ID NO:142; VH CD3 contains (i) CDR1 VH CD3 containing the amino acid sequence of SEQ ID NO:2, (ii) CDR2 VH CD3 containing the amino acid sequence of SEQ ID NO:10, and (iii) CDR3 VH CD3 containing the amino acid sequence of SEQ ID NO:4; And VL CD3 contains (i) CDR1 VL CD3 containing the amino acid sequence of SEQ ID NO:91, (ii) CDR2 VL CD3 containing the amino acid sequence of SEQ ID NO:78, and (iii) CDR3 VL CD3 containing the amino acid sequence of SEQ ID NO:79. 17. Bispecific antibody according to claim 10, where: a. VH GUCY2c contains the amino acid sequence SEQ ID NO:73; And b. VL GUCY2c contains the amino acid sequence of SEQ ID NO:147. 18. Bispecific antibody according to claim 10, where: a. VH CD3 contains the amino acid sequence SEQ ID NO:9; And b. VL CD3 contains the amino acid sequence of SEQ ID NO:90. With. VH GUCY2c contains the amino acid sequence SEQ ID NO:73; And d. VL GUCY2c contains the amino acid sequence of SEQ ID NO:147. 19. A bispecific antibody that specifically binds to GUCY2c and CD3, wherein the bispecific antibody comprises a first polypeptide chain and a second polypeptide chain, and where: a. The first polypeptide chain contains the following regions in the following order in the direction from N-terminus to C-terminus: VL antibodies against GUCY2c (VL GUCY2c) - Linker 1 - VH antibodies against CD3 (VH CD3) - Linker 3 - the first heterodimerization promoting domain, where VL GUCY2c contains the amino acid sequence SEQ ID NO:147, Linker 1 contains the amino acid sequence SEQ ID NO:190, VH CD3 contains the amino acid sequence SEQ ID NO:9, Linker 3 contains the amino acid sequence of SEQ ID NO:192, and the first heterodimerization promoting domain contains the amino acid sequence of SEQ ID NO:188; And b. the second polypeptide chain contains the following regions in the following order in the direction from N-terminus to C-terminus: VL antibodies against CD3 (VL CD3) - Linker 2 - VH antibodies against GUCY2c (VH GUCY2c) - Linker 3 - second heterodimerization promoting domain, where VL CD3 contains the amino acid sequence SEQ ID NO:90, Linker 2 contains the amino acid sequence SEQ ID NO:191, VH GUCY2c contains the amino acid sequence SEQ ID NO:73, Linker 3 contains the amino acid sequence of SEQ ID NO:192 and the second heterodimerization promoting domain contains the amino acid sequence of SEQ ID NO:189. 20. Bispecific antibody according to claim 19, where VL GUCY2c and VH GUCY2c form a domain that specifically binds to GUCY2c; And VL CD3 and VH CD3 form a domain that specifically binds to CD3; where Linker 3 of the first polypeptide chain and Linker 3 of the second polypeptide chain are covalently linked to each other by two disulfide bonds; wherein each of the first heterodimerization promoting domain and the second heterodimerization promoting domain comprises a CH2 domain and a CH3 domain; wherein the CH3 domain of the first heterodimerization promoting domain forms a protrusion, and the CH3 domain of the second heterodimerization promoting domain forms a recess; And wherein at least one disulfide bond is formed between the CH3 domain of the first heterodimerization promoting domain and the CH3 domain of the second heterodimerization promoting domain. 21. The bispecific antibody according to claim 10, wherein the bispecific antibody is a humanized antibody. 22. A bispecific antibody that specifically binds to GUCY2c and to CD3, containing a first polypeptide chain and a second polypeptide chain, wherein the first polypeptide chain contains the amino acid sequence of SEQ ID NO:216, and the second polypeptide chain contains the amino acid sequence of SEQ ID NO:220. 23. A method of treating a GUCY2c-related disease in a patient, comprising administering to the patient an antibody according to claim 1, a bispecific antibody according to claim 10, or a pharmaceutical composition containing such an antibody or bispecific antibody. 24. A nucleic acid molecule encoding an antibody according to claim 1. 25. A nucleic acid molecule encoding a bispecific antibody according to claim 10. 26. An expression vector containing the polynucleotide according to claim 24. 27. An expression vector containing the polynucleotide according to claim 25. 28. A host cell for producing an antibody according to claim 1, containing a vector according to claim 26. 29. A host cell for producing a bispecific antibody according to claim 10, containing a vector according to claim 27. 30. A method for producing an antibody, comprising culturing a host cell according to claim 28 under conditions leading to the production of the antibody according to claim 1, and purifying the antibody from the culture supernatant. 31. A method for producing a bispecific antibody, comprising culturing a host cell according to claim 29 under conditions leading to the production of a bispecific antibody according to claim 10, and purifying the bispecific antibody from the culture supernatant. 32. A composition for treating a disease associated with GUCY2c, comprising an effective amount of a bispecific antibody according to claim 10 and a second therapeutic agent. 33. A method of treating a disease associated with GUCY2c in a patient, comprising administering to the patient a combination therapy that includes the composition of claim 32. 34. A composition for the treatment of a disease associated with GUCY2c, containing an effective amount of the bispecific antibody according to claim 10 and an antidiarrheal agent. 35. A method of treating a disease associated with GUCY2c in an individual, comprising administering to the individual a combination therapeutic agent comprising the composition of claim 34. 36. A pharmaceutical composition for the treatment of a disease associated with GUCY2c, containing an effective amount of a bispecific antibody according to claim 10 and a pharmaceutically acceptable carrier.

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