Binding molecules specific for CD73 and uses thereof

Abstract

The present invention provides binding molecules specific for CD73 and uses thereof. Specifically, the present invention provides isolated antibodies or antigen-binding portions thereof that bind to CD73 and inhibit CD73 activity. The present invention also provides uses of the isolated antibodies or antigen-binding portions thereof of the present invention in treating diseases. The present invention also provides nucleic acid molecules encoding the isolated antibodies or antigen-binding portions thereof of the present invention, expression vectors, host cells and preparation methods for expressing the isolated antibodies or antigen-binding portions thereof of the present invention.

Description

Technical Field

[0001] The present invention relates to an isolated antibody or antigen-binding portion thereof that specifically binds to CD73, as well as uses of the isolated antibody or antigen-binding portion thereof of the present invention in treating diseases, and therapeutic methods using the isolated antibody or antigen-binding portion thereof of the present invention. Background of the Invention

[0003] CD73 (ecto-5'-nucleotidase) is a membrane-bound / free protein of about 70 kDa, consisting of two subunits at the N-terminus and the C-terminus and a flexible connection. The C-terminus is anchored to the cell membrane through glycosylphosphatidylinositol (GPI). CD73 exerts its enzymatic activity by switching between the open and closed states of the two subunits at the N-terminus and the C-terminus (Knapp, K. et al. (2012), Structure, 20(12), 2161-2173). In the adenosine pathway, CD73 can further hydrolyze the AMP produced by CD39 hydrolysis into adenosine. Adenosine exerts a variety of immunosuppressive effects, such as inhibiting the proliferation of CD4+T and CD8+T, inhibiting the activity of NK cells, promoting the proliferation of Treg, and so on.

[0004] Numerous reports have shown that CD73 is highly expressed in a variety of tumor tissues, including gastric cancer, triple-negative breast cancer, non-small cell adenocarcinoma, rectal adenocarcinoma, colorectal cancer, renal cancer, ovarian cancer, prostate cancer, oral squamous cell carcinoma, head and neck squamous cell carcinoma, and indicates a poor prognosis (Vijayan, D. et al., (2017), Nature reviews Cancer, 17, 709). Enhanced expression of CD73 in the tumor microenvironment is associated with tumor proliferation, metastasis, angiogenesis, and short patient survival (Allard, B. et al., (2017), Immunological reviews, 276 (1), 121-144). Along with high expression of CD73, increased adenosine inhibits tumor immunity by activating tumor-intrinsic and host-mediated tumor-promoting mechanisms, limiting immune cell infiltration, limiting cytotoxicity and the production of cytokines (such as interferon), causing strong immunosuppression (Ohta, A. et al., (2016), Front Immunol, 7, 109). Immunosuppression is a typical feature of cancer, and overcoming inhibitory barriers is crucial. Therefore, inhibiting CD73 to reduce the production of adenosine in the tumor microenvironment, thereby activating immunity and inhibiting tumor growth is a very promising treatment direction.

[0005] Antibodies can inhibit the enzymatic activity of CD73 by antagonizing the conformational change of CD73 to the active form, but there is no report of antibodies directly binding to the active site of CD73 to inhibit its enzymatic activity. At the same time, although CD73 is a very potential therapeutic target, more research is still needed to determine the application prospects of the CD73 target and develop related drugs and combination therapies. Summary of the invention

[0006] The present invention provides isolated antibodies or antigen-binding portions that specifically bind to CD73 and their use in treating diseases.

[0007] In one aspect, the present invention provides an isolated antibody or antigen-binding portion thereof, wherein the isolated antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising HCDR1, HCDR2, HCDR3 sequences; and a light chain variable region comprising LCDR1, LCDR2, LCDR3 sequences, wherein:

[0008] (a) the HCDR1 comprises an amino acid sequence selected from SEQ ID NOs: 5, 18, 31, 43 and 56 and conservative modifications thereof;

[0009] (b) the HCDR2 comprises an amino acid sequence selected from SEQ ID NOs: 6, 19, 32, 44 and 57 and conservative modifications thereof;

[0010] (c) the HCDR3 comprises an amino acid sequence selected from SEQ ID NOs: 7, 20, 33, 45 and 58 and conservative modifications thereof;

[0011] (d) the LCDR1 comprises an amino acid sequence selected from SEQ ID NOs: 10, 23, 36, 48 and 61 and conservatively modified forms thereof;

[0012] (e) the LCDR2 comprises an amino acid sequence selected from SEQ ID NOs: 11, 24, 37, 49 and 62 and conservative modifications thereof; and

[0013] (f) The LCDR3 comprises an amino acid sequence selected from SEQ ID NOs: 12, 25, 38, 50 and 63 and conservatively modified forms thereof.

[0014] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises:

[0015] 1) (a) comprising HCDR1 of SEQ ID NO: 5, (b) comprising HCDR2 of SEQ ID NO: 6, (c) comprising HCDR3 of SEQ ID NO: 7, (d) comprising LCDR1 of SEQ ID NO: 10, (e) comprising LCDR2 of SEQ ID NO: 11, and (f) comprising LCDR3 of SEQ ID NO: 12;

[0016] 2) (a) comprising HCDR1 of SEQ ID NO: 18, (b) comprising HCDR2 of SEQ ID NO: 19, (c) comprising HCDR3 of SEQ ID NO: 20, (d) comprising LCDR1 of SEQ ID NO: 23, (e) comprising LCDR2 of SEQ ID NO: 24, and (f) comprising LCDR3 of SEQ ID NO: 25;

[0017] 3) (a) HCDR1 comprising SEQ ID NO:31, (b) HCDR2 comprising SEQ ID NO:32, (c) HCDR3 comprising SEQ ID NO:33, (d) LCDR1 comprising SEQ ID NO:36, (e) LCDR2 comprising SEQ ID NO:37, and (f) LCDR3 comprising SEQ ID NO:38;

[0018] 4) (a) comprising HCDR1 of SEQ ID NO:43, (b) comprising HCDR2 of SEQ ID NO:44, (c) comprising HCDR3 of SEQ ID NO:45, (d) comprising LCDR1 of SEQ ID NO:48, (e) comprising LCDR2 of SEQ ID NO:49, and (f) comprising LCDR3 of SEQ ID NO:50; or

[0019] 5) (a) HCDR1 comprising SEQ ID NO:56, (b) HCDR2 comprising SEQ ID NO:57, (c) HCDR3 comprising SEQ ID NO:58, (d) LCDR1 comprising SEQ ID NO:61, (e) LCDR2 comprising SEQ ID NO:62, and (f) LCDR3 comprising SEQ ID NO:63.

[0020] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises: (a) HCDR1 comprising SEQ ID NO: 5, (b) HCDR2 comprising SEQ ID NO: 6, (c) HCDR3 comprising SEQ ID NO: 7, (d) LCDR1 comprising SEQ ID NO: 10, (e) LCDR2 comprising SEQ ID NO: 11, and (f) LCDR3 comprising SEQ ID NO: 12.

[0021] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises: (a) HCDR1 comprising SEQ ID NO: 18, (b) HCDR2 comprising SEQ ID NO: 19, (c) HCDR3 comprising SEQ ID NO: 20, (d) LCDR1 comprising SEQ ID NO: 23, (e) LCDR2 comprising SEQ ID NO: 24, and (f) LCDR3 comprising SEQ ID NO: 25.

[0022] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises: (a) HCDR1 comprising SEQ ID NO: 31, (b) HCDR2 comprising SEQ ID NO: 32, (c) HCDR3 comprising SEQ ID NO: 33, (d) LCDR1 comprising SEQ ID NO: 36, (e) LCDR2 comprising SEQ ID NO: 37, and (f) LCDR3 comprising SEQ ID NO: 38.

[0023] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises: (a) HCDR1 comprising SEQ ID NO: 43, (b) HCDR2 comprising SEQ ID NO: 44, (c) HCDR3 comprising SEQ ID NO: 45, (d) LCDR1 comprising SEQ ID NO: 48, (e) LCDR2 comprising SEQ ID NO: 49, and (f) LCDR3 comprising SEQ ID NO: 50.

[0024] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises: (a) HCDR1 comprising SEQ ID NO: 56, (b) HCDR2 comprising SEQ ID NO: 57, (c) HCDR3 comprising SEQ ID NO: 58, (d) LCDR1 comprising SEQ ID NO: 61, (e) LCDR2 comprising SEQ ID NO: 62, and (f) LCDR3 comprising SEQ ID NO: 63.

[0025] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises:

[0026] (i) the heavy chain variable region (VH) comprises an amino acid sequence having at least 85% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 16, 29, 41, 54, 66, 67, 76, 77 and 78 and conservatively modified versions thereof; and

[0027] (ii) the light chain variable region (VL) comprises an amino acid sequence having at least 85% homology to an amino acid sequence selected from the following group: SEQ ID NOs: 8, 21, 34, 46, 59, 68, 69, 70, 79 and 80 and conservatively modified versions thereof.

[0028] In certain embodiments, the heavy chain variable region comprises an amino acid sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homologous to the heavy chain variable region selected from (i); the light chain variable region comprises an amino acid sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homologous to the light chain variable region selected from (ii).

[0029] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises:

[0030] 1) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acid sequence of SEQ ID NO: 3, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acid sequence of SEQ ID NO: 8;

[0031] 2) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 16, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 21;

[0032] 3) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 29, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 34;

[0033] 4) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 41, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 46;

[0034] 5) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 54, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 59;

[0035] 6) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 66, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 68;

[0036] 7) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 66, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 69;

[0037] 8) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 66, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 70;

[0038] 9) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 67, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 68;

[0039] 10) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 67, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 69;

[0040] 11) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 67, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 70;

[0041] 12) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 76, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 79;

[0042] 13) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 76, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 80;

[0043] 14) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 77, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 79;

[0044] 15) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 77, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 80;

[0045] 16) a heavy chain variable region (VH) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 78, and a light chain variable region (VL) comprising an amino acid sequence at least 85% homologous to the amino acids of SEQ ID NO: 79;

[0046] 17) a heavy chain variable region (VH) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 78, and a light chain variable region (VL) comprising an amino acid sequence that is at least 85% homologous to the amino acids of SEQ ID NO: 80.

[0047] In certain embodiments, the above-mentioned isolated antibody or antigen-binding portion thereof, wherein the above-mentioned heavy chain variable region and light chain variable region respectively comprise an amino acid sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homologous to the heavy chain variable region and light chain variable region selected from 1)-17).

[0048] In certain embodiments, the isolated antibody described above is IgG.

[0049] In certain embodiments, the isolated antibody described above is IgG1, IgG2 or IgG4.

[0050] In certain embodiments, the isolated antibody is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human modified antibody, a human antibody, a Fv, a single chain antibody (scFv), a Fab, a Fab', a Fab'-SH or a F(ab') 2 .

[0051] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises a heavy chain and a light chain, wherein:

[0052] (I) the heavy chain comprises an amino acid sequence having at least 85% homology to an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 15, 26, 27, 39, 51, 52, 64, 71, 72, 81, 82, 83, 86, 87, 90, 91, 124, 125, 126 and 127 and conservatively modified versions thereof; and

[0053] (II) The light chain comprises an amino acid sequence having at least 85% homology to an amino acid sequence selected from the following group: SEQ ID NOs: 14, 28, 40, 53, 65, 73, 74, 75, 84, 85, 88 and 89 and conservatively modified versions thereof.

[0054] In certain embodiments, the heavy chain comprises an amino acid sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homologous to the heavy chain selected from (I); the light chain comprises an amino acid sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homologous to the light chain selected from (II).

[0055] In certain embodiments, the isolated antibody or antigen-binding portion thereof described above comprises:

[0056] 1) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 13, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 14;

[0057] 2) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 15, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 14;

[0058] 3) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 26, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 28;

[0059] 4) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 27, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 28;

[0060] 5) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 39, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 40;

[0061] 6) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 51, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 53;

[0062] 7) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 52, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 53;

[0063] 8) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 64, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 65;

[0064] 9) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 71, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 73;

[0065] 10) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 71, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 74;

[0066] 11) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 71, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 75;

[0067] 12) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 72, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 73;

[0068] 13) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 72, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 74;

[0069] 14) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 72, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 75;

[0070] 15) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 81, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0071] 16) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 81, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85;

[0072] 17) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 82, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0073] 18) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 82, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85;

[0074] 19) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 83, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0075] 20) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 83, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85;

[0076] 21) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 90, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0077] 22) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 90, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85;

[0078] 23) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 91, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0079] 24) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 91, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85;

[0080] 25) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 86, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 88;

[0081] 26) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 86, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 89;

[0082] 27) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 87, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 88;

[0083] 28) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 87, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 89;

[0084] 29) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 124, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0085] 30) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 124, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85;

[0086] 31) a heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 125, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 84;

[0087] 32) A heavy chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 125, and a light chain comprising an amino acid sequence at least 85% homologous to the amino acid sequence of SEQ ID NO: 85.

[0088] In certain embodiments, the heavy chain and light chain respectively comprise an amino acid sequence that is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homologous to the heavy chain and light chain selected from 1)-32).

[0089] In another aspect, the invention provides an isolated antibody or antigen-binding portion thereof comprising the heavy chain set forth in SEQ ID NO:71 and the light chain set forth in SEQ ID NO:73.

[0090] In yet another aspect, the invention provides an isolated antibody, or antigen-binding portion thereof, comprising a heavy chain consisting essentially of SEQ ID NO:72, and a light chain consisting essentially of SEQ ID NO:74.

[0091] In yet another aspect, the invention provides an isolated antibody or antigen-binding portion thereof comprising the heavy chain set forth in SEQ ID NO:72 and the light chain set forth in SEQ ID NO:75.

[0092] In another aspect, the invention provides an isolated antibody, or antigen-binding portion thereof, comprising a heavy chain consisting essentially of SEQ ID NO:81, and a light chain consisting essentially of SEQ ID NO:85.

[0093] In yet another aspect, the invention provides an isolated antibody, or antigen-binding portion thereof, comprising a heavy chain consisting essentially of SEQ ID NO:82, and a light chain consisting essentially of SEQ ID NO:84.

[0094] In yet another aspect, the invention provides an isolated antibody, or antigen-binding portion thereof, comprising a heavy chain consisting essentially of SEQ ID NO:83, and a light chain consisting essentially of SEQ ID NO:85.

[0095] In another aspect, the invention provides an isolated antibody, or antigen-binding portion thereof, comprising a heavy chain consisting essentially of SEQ ID NO: 124, and a light chain consisting essentially of SEQ ID NO: 85.

[0096] In yet another aspect, the invention provides an isolated antibody, or antigen-binding portion thereof, comprising a heavy chain consisting essentially of SEQ ID NO: 125, and a light chain consisting essentially of SEQ ID NO: 85.

[0097] In certain embodiments, the above-mentioned isolated antibody or antigen-binding portion thereof is a CD73 antagonist or a 5' nucleosidase antagonist of CD73.

[0098] In certain embodiments, the above-mentioned CD73 is human CD73.

[0099] In another aspect, the present invention provides a method for reducing adenosine levels in a subject having a tumor, comprising administering to the subject an effective amount of the above-described isolated antibody or antigen-binding portion thereof.

[0100] In yet another aspect, the present invention provides a method for enhancing T cell responses in a subject having a tumor, comprising administering an effective dose of the above-described isolated antibody or antigen-binding portion thereof.

[0101] In yet another aspect, the present invention provides a method for stimulating an immune response in a subject, comprising administering to the subject an effective dose of the above-described isolated antibody or antigen-binding portion thereof.

[0102] In certain embodiments, the subject is a subject suffering from a tumor.

[0103] In certain embodiments, the subject described above has tumor cells expressing CD73 and / or a tumor microenvironment containing CD73.

[0104] In another aspect, the present invention provides a method for inhibiting the growth of tumor cells in a subject, comprising administering to the subject an effective dose of the above-described isolated antibody or antigen-binding portion thereof.

[0105] In another aspect, the present invention provides use of the above-mentioned isolated antibody or antigen-binding portion thereof in the preparation of a medicament for reducing adenosine levels in tumor cells and / or tumor microenvironment.

[0106] In yet another aspect, the present invention provides a use of the above-mentioned isolated antibody or antigen-binding portion thereof in the preparation of a medicament for stimulating a T cell response in a subject suffering from a tumor.

[0107] In another aspect, the present invention provides use of the isolated antibody or antigen-binding portion thereof described above in the preparation of a medicament for stimulating an immune response in a subject.

[0108] In certain embodiments, the subject is a subject suffering from a tumor.

[0109] In certain embodiments, the subject described above has tumor cells expressing CD73 and / or a tumor microenvironment containing CD73.

[0110] In another aspect, the present invention provides use of the above-mentioned isolated antibody or antigen-binding portion thereof in the preparation of a medicament for inhibiting the growth of tumor cells in a subject.

[0111] In yet another aspect, the present invention provides the above-described isolated antibody or antigen-binding portion thereof for use in reducing adenosine levels in tumor cells and / or tumor microenvironment.

[0112] In yet another aspect, the present invention provides the above-described isolated antibody or antigen-binding portion thereof for use in stimulating a T cell response in an individual suffering from a tumor.

[0113] In another aspect, the invention provides the isolated antibody or antigen-binding portion thereof as described above for use in stimulating an immune response in a subject.

[0114] In certain embodiments, the subject described above is a subject suffering from a tumor.

[0115] In certain embodiments, the subject described above has tumor cells expressing CD73 and / or a tumor microenvironment containing CD73.

[0116] In another aspect, the present invention provides the isolated antibody or antigen-binding portion thereof described above for use in inhibiting the growth of tumor cells in a subject.

[0117] In yet another aspect, the present invention provides an isolated nucleic acid composition comprising:

[0118] (I) a first nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 4, 17, 30, 42, 55, 92, 93, 102, 103 and 104, wherein the first nucleic acid encodes a heavy chain variable region (VH) of the isolated antibody or antigen-binding portion thereof; and

[0119] (II) a second nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 9, 22, 35, 47, 60, 94, 95, 96, 105 and 106, wherein the second nucleic acid encodes the light chain variable region (VL) of the isolated antibody or antigen-binding portion thereof.

[0120] In certain embodiments, the isolated nucleic acid composition described above comprises:

[0121] 1) containing the first nucleic acid shown in SEQ ID NO: 4, and containing the second nucleic acid shown in SEQ ID NO: 9;

[0122] 2) comprising the first nucleic acid shown in SEQ ID NO: 17, and comprising the second nucleic acid shown in SEQ ID NO: 22;

[0123] 3) comprising the first nucleic acid shown in SEQ ID NO: 30, and comprising the second nucleic acid shown in SEQ ID NO: 35;

[0124] 4) comprising the first nucleic acid shown in SEQ ID NO: 42, and comprising the second nucleic acid shown in SEQ ID NO: 47;

[0125] 5) comprising the first nucleic acid shown in SEQ ID NO: 55, and comprising the second nucleic acid shown in SEQ ID NO: 60;

[0126] 6) comprising the first nucleic acid shown in SEQ ID NO: 92, and comprising the second nucleic acid shown in SEQ ID NO: 94;

[0127] 7) comprising the first nucleic acid shown in SEQ ID NO: 92, and comprising the second nucleic acid shown in SEQ ID NO: 95;

[0128] 8) comprising the first nucleic acid shown in SEQ ID NO: 92, and comprising the second nucleic acid shown in SEQ ID NO: 96;

[0129] 9) comprising the first nucleic acid shown in SEQ ID NO: 93, and comprising the second nucleic acid shown in SEQ ID NO: 94;

[0130] 10) comprising the first nucleic acid shown in SEQ ID NO: 93, and comprising the second nucleic acid shown in SEQ ID NO: 95;

[0131] 11) comprising the first nucleic acid shown in SEQ ID NO: 93, and comprising the second nucleic acid shown in SEQ ID NO: 96;

[0132] 12) comprising the first nucleic acid shown in SEQ ID NO: 102, and comprising the second nucleic acid shown in SEQ ID NO: 105;

[0133] 13) comprising the first nucleic acid shown in SEQ ID NO: 102, and comprising the second nucleic acid shown in SEQ ID NO: 106;

[0134] 14) comprising the first nucleic acid shown in SEQ ID NO: 103, and comprising the second nucleic acid shown in SEQ ID NO: 105;

[0135] 15) comprising the first nucleic acid shown in SEQ ID NO: 103, and comprising the second nucleic acid shown in SEQ ID NO: 106;

[0136] 16) comprising the first nucleic acid shown in SEQ ID NO: 104, and comprising the second nucleic acid shown in SEQ ID NO: 105;

[0137] 17) comprising the first nucleic acid shown in SEQ ID NO: 104, and comprising the second nucleic acid shown in SEQ ID NO: 106.

[0138] In another aspect, the present invention provides an expression vector composition comprising:

[0139] (I) a first expression vector, wherein the first expression vector comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 4, 17, 30, 42, 55, 92, 93, 102, 103 and 104; and

[0140] (II) A second expression vector, wherein the second expression vector comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 9, 22, 35, 47, 60, 94, 95, 96, 105 and 106.

[0141] In certain embodiments, the above-mentioned expression vector composition comprises:

[0142] 1) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 4; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 9;

[0143] 2) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 17; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 22;

[0144] 3) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 30; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 35;

[0145] 4) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 42; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 47;

[0146] 5) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 55; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 60;

[0147] 6) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 92; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 94;

[0148] 7) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 92; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 95;

[0149] 8) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 92; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 96;

[0150] 9) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 93; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 94;

[0151] 10) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 93; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 95;

[0152] 11) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 93; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 96;

[0153] 12) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 102; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 105;

[0154] 13) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 102; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 106;

[0155] 14) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 103; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 105;

[0156] 15) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 103; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 106;

[0157] 16) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 104; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 105; or

[0158] 17) A first expression vector, wherein the first expression vector contains the nucleotide sequence shown in SEQ ID NO: 104; and a second expression vector, wherein the second expression vector contains the nucleotide sequence shown in SEQ ID NO: 106.

[0159] In another aspect, the present invention provides an expression vector comprising:

[0160] (I) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 4, 17, 30, 42, 55, 92, 93, 102, 103 and 104; and

[0161] (II) a second nucleic acid sequence, wherein the second nucleic acid sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 9, 22, 35, 47, 60, 94, 95, 96, 105 and 106.

[0162] In certain embodiments, the above-mentioned expression vector comprises:

[0163] 1) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 4; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 9;

[0164] 2) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 17; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 22;

[0165] 3) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 30; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 35;

[0166] 4) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 42; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 47;

[0167] 5) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 55; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 60;

[0168] 6) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 92; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 94;

[0169] 7) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 92; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 95;

[0170] 8) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 92; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 96;

[0171] 9) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 93; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 94;

[0172] 10) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 93; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 95;

[0173] 11) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 93; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 96;

[0174] 12) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 102; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 105;

[0175] 13) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 102; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 106;

[0176] 14) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 103; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 105;

[0177] 15) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 103; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 106;

[0178] 16) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 104; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 105; or

[0179] 17) A first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 104; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 106.

[0180] In another aspect, the present invention provides an isolated nucleic acid composition comprising:

[0181] (I) a first nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 97, 98, 107, 108, 109, 112, 113, 116 and 117, wherein the first nucleic acid encodes a heavy chain of the isolated antibody or antigen-binding portion thereof; and

[0182] (II) a second nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 99, 100, 101, 110, 111, 114 and 115, wherein the second nucleic acid encodes the light chain of the isolated antibody or antigen-binding portion thereof.

[0183] In certain embodiments, the isolated nucleic acid composition described above comprises:

[0184] 1) comprising the first nucleic acid shown in SEQ ID NO: 97, and comprising the second nucleic acid shown in SEQ ID NO: 99;

[0185] 2) comprising the first nucleic acid shown in SEQ ID NO: 97, and comprising the second nucleic acid shown in SEQ ID NO: 100;

[0186] 3) comprising the first nucleic acid shown in SEQ ID NO: 97, and comprising the second nucleic acid shown in SEQ ID NO: 101;

[0187] 4) comprising the first nucleic acid shown in SEQ ID NO: 98, and comprising the second nucleic acid shown in SEQ ID NO: 99;

[0188] 5) comprising the first nucleic acid shown in SEQ ID NO: 98, and comprising the second nucleic acid shown in SEQ ID NO: 100;

[0189] 6) comprising the first nucleic acid shown in SEQ ID NO: 98, and comprising the second nucleic acid shown in SEQ ID NO: 101;

[0190] 7) comprising the first nucleic acid shown in SEQ ID NO: 107, and comprising the second nucleic acid shown in SEQ ID NO: 110;

[0191] 8) comprising the first nucleic acid shown in SEQ ID NO: 107, and comprising the second nucleic acid shown in SEQ ID NO: 111;

[0192] 9) comprising the first nucleic acid shown in SEQ ID NO: 108, and comprising the second nucleic acid shown in SEQ ID NO: 110;

[0193] 10) comprising the first nucleic acid shown in SEQ ID NO: 108, and comprising the second nucleic acid shown in SEQ ID NO: 111;

[0194] 11) comprising the first nucleic acid shown in SEQ ID NO: 109, and comprising the second nucleic acid shown in SEQ ID NO: 110;

[0195] 12) comprising the first nucleic acid shown in SEQ ID NO: 109, and comprising the second nucleic acid shown in SEQ ID NO: 111;

[0196] 13) comprising the first nucleic acid shown in SEQ ID NO: 116, and comprising the second nucleic acid shown in SEQ ID NO: 110;

[0197] 14) comprising the first nucleic acid shown in SEQ ID NO: 116, and comprising the second nucleic acid shown in SEQ ID NO: 111;

[0198] 15) comprising the first nucleic acid shown in SEQ ID NO: 117, and comprising the second nucleic acid shown in SEQ ID NO: 110;

[0199] 16) comprising the first nucleic acid shown in SEQ ID NO: 117, and comprising the second nucleic acid shown in SEQ ID NO: 111;

[0200] 17) comprising the first nucleic acid shown in SEQ ID NO: 112, and comprising the second nucleic acid shown in SEQ ID NO: 114;

[0201] 18) comprising the first nucleic acid shown in SEQ ID NO: 112, and comprising the second nucleic acid shown in SEQ ID NO: 115;

[0202] 19) comprising the first nucleic acid shown in SEQ ID NO: 113, and comprising the second nucleic acid shown in SEQ ID NO: 114;

[0203] 20) comprising the first nucleic acid shown in SEQ ID NO: 113, and comprising the second nucleic acid shown in SEQ ID NO: 115.

[0204] In another aspect, the present invention provides an expression vector composition comprising:

[0205] (I) a first expression vector, wherein the first expression vector comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 97, 98, 107, 108, 109, 112, 113, 116 and 117; and

[0206] (II) a second expression vector, wherein the second expression vector comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 99, 100, 101, 110, 111, 114 and 115.

[0207] In certain embodiments, the above-mentioned expression vector composition comprises:

[0208] 1) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 97; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 99;

[0209] 2) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 97; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 100;

[0210] 3) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 97; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 101;

[0211] 4) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 98; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 99;

[0212] 5) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 98; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 100;

[0213] 6) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 98; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 101;

[0214] 7) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 107; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0215] 8) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 107; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0216] 9) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 108; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0217] 10) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 108; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0218] 11) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 109; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0219] 12) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 109; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0220] 13) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 116; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0221] 14) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 116; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0222] 15) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 117; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0223] 16) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 117; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0224] 17) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 112; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 114;

[0225] 18) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 112; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 115;

[0226] 19) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 113; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 114; or

[0227] 20) A first expression vector, wherein the first expression vector contains the nucleotide sequence shown in SEQ ID NO: 113; and a second expression vector, wherein the second expression vector contains the nucleotide sequence shown in SEQ ID NO: 115.

[0228] In another aspect, the present invention provides an expression vector comprising:

[0229] (I) a first nucleic acid sequence comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 97, 98, 107, 108, 109, 112, 113, 116 and 117; and

[0230] (II) A second nucleic acid sequence comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 99, 100, 101, 110, 111, 114 and 115.

[0231] In certain embodiments, the above-mentioned expression vector comprises:

[0232] 1) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 97; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 99;

[0233] 2) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 97; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 100;

[0234] 3) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 97; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 101;

[0235] 4) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 98; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 99;

[0236] 5) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 98; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 100;

[0237] 6) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 98; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 101;

[0238] 7) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 107; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0239] 8) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 107; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0240] 9) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 108; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0241] 10) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 108; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0242] 11) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 109; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0243] 12) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 109; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0244] 13) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 116; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0245] 14) a first nucleic acid sequence vector, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 116; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0246] 15) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 117; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 110;

[0247] 16) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 117; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 111;

[0248] 17) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 112; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 114;

[0249] 18) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 112; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 115;

[0250] 19) a first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 113; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 114; or

[0251] 20) A first nucleic acid sequence, wherein the first nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 113; and a second nucleic acid sequence, wherein the second nucleic acid sequence comprises the nucleotide sequence shown in SEQ ID NO: 115.

[0252] In another aspect, the present invention provides a cell comprising any one of the above-mentioned expression vector compositions.

[0253] In another aspect, the present invention provides a method for preparing an isolated antibody or an antigen-binding portion thereof, comprising: expressing the isolated antibody or the antigen-binding portion thereof in the above-mentioned cells, and isolating the isolated antibody or the antigen-binding portion thereof from the above-mentioned cells.

[0254] In another aspect, the present invention provides a pharmaceutical composition comprising the isolated antibody or antigen-binding portion thereof described above, and a pharmaceutically acceptable carrier.

[0255] In yet another aspect, the present invention provides a kit comprising the isolated antibody or antigen-binding portion thereof described above.

[0256] On the other hand, the present invention provides a method for treating a tumor, comprising administering an effective dose of the above-mentioned isolated antibody or its antigen-binding fragment, or the above-mentioned pharmaceutical composition to a subject in need. In some embodiments, the tumor is selected from a solid tumor or a hematological tumor. In some embodiments, the tumor is selected from bladder cancer, breast cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer (small cell carcinoma or non-small cell lung cancer), gastric cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, central nervous system tumors, lymphoma, leukemia, myeloma, sarcoma and virus-related cancer.

[0257] In another aspect, the present invention provides the use of the above-mentioned antibody or its antigen-binding fragment, or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating a tumor. In some embodiments, the tumor is selected from a solid tumor or a hematological tumor. In some embodiments, the tumor is selected from bladder cancer, breast cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer (small cell carcinoma or non-small cell lung cancer), gastric cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, central nervous system tumors, lymphoma, leukemia, myeloma, sarcoma and virus-related cancer.

[0258] In another aspect, the present invention provides the above-mentioned antibody or its antigen-binding fragment or the above-mentioned pharmaceutical composition for treating a tumor. In some embodiments, the cancer is selected from a solid tumor or a hematological tumor. In some embodiments, the cancer is selected from bladder cancer, breast cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer (small cell carcinoma or non-small cell lung cancer), gastric cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, central nervous system tumors, lymphoma, leukemia, myeloma, sarcoma and virus-related cancer.

[0259] Table I. Description of antibody sequences of the present invention

[0260]

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293] Note: All amino acid numbers in CDR and framework regions are annotated according to the EU index of the Kabat system (Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, 5th edition, US Department of Health and Human Services, NIH publication No. 91-3242). All sequences do not include signal peptides. aa: amino acid, nt: nucleotide. BRIEF DESCRIPTION OF THE DRAWINGS

[0294] Figure 1 The binding ability of positive clones to soluble huCD73 protein was determined;

[0295] Figure 2 The binding ability of positive clones to natural CD73 protein was determined;

[0296] Figure 3 Shows blockade of soluble recombinant human CD73 enzyme activity;

[0297] Figure 4 Shows cell-based enzyme activity blocking assay;

[0298] Figure 5 CD4+ T cell proliferation reversed by CD73 antibody was shown;

[0299] Figure 6 Shows CD73 enzyme activity of CD4+ T cells blocked by antibodies;

[0300] Figure 7 CD4 + T cell IFN-γ release was shown to be reversed by antibodies;

[0301] Figure 8 Demonstrating CD73 antibody-mediated CD73 endocytosis;

[0302] Fig. 9 Show the affinity of humanized antibodies and chimeric antibodies detected by ELISA;

[0303] Fig.10 Show the affinity of humanized and chimeric antibodies detected by FACs;

[0304] Fig.11 Cell surface 5′ ectonucleotidase activity blocked by humanized antibodies was shown;

[0305] Fig.12 Shows the 5'ectonucleotidase activity of U87-MG cells blocked by humanized antibodies;

[0306] Fig.13 Showing CD4+ T cell proliferation reversed by humanized CD73 antibody;

[0307] Fig.14 Showing that humanized CD73 antibody reversed IFN-γ release from CD4+ T cells;

[0308] Fig.15A and Fig. 15B The humanized antibody was shown to inhibit the 5' ectonucleotidase activity of xenograft tumors in animal models.

[0309] Fig.16 The humanized antibody showed an anti-tumor effect in the A375 human melanoma xenograft model.

[0310] Detailed description

[0311] Described herein are isolated antibodies, specifically monoclonal antibodies, such as human monoclonal antibodies ("antagonistic anti-CD73 antibodies"), that specifically bind to CD73 and thereby reduce CD73 activity. In certain embodiments, the antibodies described herein are derived from specific heavy and light chain germline sequences and / or comprise specific structural features, such as CDR regions, including specific amino acid sequences. Provided herein are isolated antibodies and methods for preparing these antibodies. Also provided herein are methods for slowing tumor growth using the antibodies alone or in combination with other therapeutic agents (e.g., antibodies) and / or cancer therapies. Therefore, the anti-CD73 antibodies described herein can be used in the treatment of numerous therapeutic applications, including, for example, inhibiting tumor growth, inhibiting metastasis, and enhancing immune responses against tumors.

[0312] definition

[0313] To make this specification easier to understand, some terms are first defined. Other definitions are set forth throughout the specification.

[0314] As used herein, the term "cluster of differentiation 73" or "CD73" refers to an enzyme (nucleotidase) that is capable of converting extracellular nucleoside 5' monophosphates into nucleosides, i.e., converting adenosine monophosphate (AMP) into adenosine. CD73 usually appears as a dimer anchored to the cell membrane via a glycosylphosphatidylinositol (GPI) bond, has extracellular enzymatic activity, and plays a role in signal transduction. The main function of CD73 is to convert extracellular nucleotides (e.g., 5'-AMP) into adenosine, which is a highly immunosuppressive molecule. Therefore, ecto-5'-nucleotidase catalyzes the dephosphorylation of purine and pyrimidine ribonucleosides and deoxyribonucleoside monophosphates into the corresponding nucleosides. Although CD73 has a wide range of substrate specificity, it prefers purine ribonucleosides.

[0315] CD73 is also known as exo-5' nuclease (5'-NT, EC 3.1.3.5). The term "CD73" includes any variant or subtype of CD73 that is naturally expressed by cells. Therefore, the antibodies described herein may cross-react with CD73 from non-human species (e.g., cynomolgus monkey CD73). Alternatively, these antibodies may be specific for human CD73 and may not exhibit any cross-reactivity with other species. CD73 or any variant and subtype thereof may be isolated from cells or tissues that naturally express it or produced recombinantly using techniques well known in the art and / or the methods described herein.

[0316] Two isoforms of human CD73 have been identified, both of which share identical N-terminal and C-terminal portions. Isoform 1 (Accession No. NP_002517.1; SEQ ID NO: 1) represents the longest protein consisting of 574 amino acids and 9 exons. Isoform 2 (Accession No. NP_001191742.1) encodes a shorter protein consisting of 524 amino acids, lacking amino acids 404-453. Isoform 2 lacks an alternative in-frame exon, resulting in a transcript with only 8 exons, but with identical N- and C-terminal sequences.

[0317] The cynomolgus (cyno) CD73 protein sequence is provided as SEQ ID NO: 2. The terms cynomolgus and cynomolgus both refer to the species Macaca fascicularis and are used interchangeably throughout this specification.

[0318] The term "antibody" as used herein may include a complete antibody and any antigen-binding fragment thereof (i.e., "antigen-binding portion") or a single chain. In one embodiment, an "antibody" refers to a glycoprotein or antigen-binding portion thereof comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. In some naturally occurring IgG, IgD, and IgA antibodies, the heavy chain constant region comprises three domains, CH1, CH2, and CH3. In some naturally occurring antibodies, each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL regions may be further subdivided into hypervariable regions, referred to as complementarity determining regions (CDRs), and more conservative regions, referred to as framework regions (FRs), which are interspersed. In this article, the CDR of the VH region is abbreviated as HCDR, that is, the three CDRs of the VH region can be abbreviated as HCDR1, HCDR2, and HCDR3; the CDR of the VL region is abbreviated as LCDR, that is, the three CDRs of the VL region can be abbreviated as LCDR1, LCDR2, and LCDR3. Each VH and VL is composed of three CDRs and four FRs, which are arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of the antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0319] The heavy chain of the antibody may or may not contain a terminal lysine (K) or a terminal glycine and lysine (GK). Thus, any heavy chain sequence and heavy chain constant region sequence provided herein may end with GK or G, or lack K or GK, regardless of the last amino acid of the sequence. This is because the terminal lysine, and sometimes glycine and lysine, are cleaved off during antibody expression.

[0320] Antibodies typically bind specifically to their cognate antigen with high affinity ranging from 10 -7 M to 10 -11 M or less dissociation constant (K D ) is reflected. It is generally considered to be greater than about 10 -6 Any K of M D Indicates non-specific binding. As used herein, an antibody that "specifically binds" to an antigen refers to an antibody that binds to the antigen with high affinity and is substantially the same antigen, but does not bind to unrelated antigens with high affinity, high affinity meaning having 10 -7 M or less, preferably 10 -8 M or less, even more preferably 5×10 -9M or less, and preferably between 10 -8 M and 10 -10 M or smaller K D An antigen is "substantially identical" to a given antigen if it exhibits a high degree of sequence identity to the given antigen, for example, if it exhibits at least 80%, at least 90%, at least 95%, at least 97%, or at least 99% or greater sequence identity to the sequence of the given antigen. For example, an antibody that specifically binds to human CD73 may also cross-react with CD73 from certain non-human primate species (e.g., cynomolgus monkeys), but may not cross-react with CD73 from other species, or with antigens other than CD73.

[0321] The immunoglobulin may be from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG, and IgM. The IgG isotype is divided into multiple subclasses in certain species: IgG1, IgG2, IgG3, and IgG4 in humans, and IgG1, IgG2a, IgG2b, and IgG3 in mice. In certain embodiments, the anti-CD73 antibodies described herein are of human IgG1 or IgG2 subtypes. Immunoglobulins (e.g., human IgG1) exist in several isotypes that differ from each other by at most a few amino acids. For example, "antibodies" may include naturally occurring and non-naturally occurring antibodies; monoclonal and polyclonal antibodies; chimeric and humanized antibodies; human and non-human antibodies; fully synthetic antibodies; and single-chain antibodies.

[0322] As used herein, the term "antigen-binding portion" of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., human CD73). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody (e.g., the anti-CD73 antibody described herein) include (i) Fab fragments, which are monovalent fragments consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab') 2(iii) a Fd fragment, which is a bivalent fragment comprising two Fab fragments connected by a disulfide bridge at the hinge region; (iv) a Fv fragment, which is composed of the VL and VH domains of one single arm of the antibody, (v) a dAb fragment (Ward et al. (1989) Nature 341:544-546), which is composed of the VH domain; and (vi) an isolated complementarity determining region (CDR), or (vii) a combination of two or more isolated CDRs, which may be optionally linked by a synthetic linker. In addition, although the two domains of the Fv fragment, VL and VH, are encoded by different genes, they can be linked to these domains by synthetic linkers using recombinant methods, allowing them to be prepared as a single protein chain, in which the VL and VH regions are paired to form a monovalent molecule (called single-chain Fv (scFv)); see, for example, Bird et al., (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). These single-chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. These and other potential constructs are described in Chan and Carter (2010) Nat. Rev. Immunol. 10:301. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and screening from these fragments can be used in the same manner as intact antibodies. Antigen-binding portions can be produced by recombinant DNA technology or by enzymatic or chemical cleavage of intact immunoglobulins.

[0323] The term "conservatively modified amino acid sequence" refers to amino acid modifications that do not significantly affect or alter the binding properties of an antibody comprising the amino acid sequence, including amino acid substitutions, additions and deletions. Modifications can be introduced into the antibodies of the invention by standard techniques such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are substitutions in which an amino acid residue is replaced by an amino acid residue with a similar side chain. Families of amino acid residues with similar side chains have been identified in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), amino acids with non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues in the CDR region of an antibody of the invention may be replaced by other amino acid residues from the same side chain family, and the retained function of the altered antibody may be tested using the functional assays described herein. Preferably, conservative modifications are no more than 1 or 2 in number.

[0324] "Bispecific" or "bifunctional antibodies" are artificial hybrid antibodies with two different heavy chain / light chain pairs, thereby generating two antigen binding sites specific for different antigens. Bispecific antibodies can be produced by a variety of methods, including hybridoma fusion or linking Fab' fragments. For example, see Songsivilai and Lachmann, Clin. Exp. Immunol. 79: 315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0325] As used herein, the term "monoclonal antibody" refers to an antibody that displays a single binding specificity and affinity for a specific epitope, or an antibody composition, wherein all antibodies in the antibody composition display a single binding specificity and affinity for a specific epitope. Typically, these monoclonal antibodies will be derived from a single antibody encoding cell or nucleic acid, and will proliferate under conditions where no sequence changes are intentionally introduced. Therefore, the term "human monoclonal antibody" refers to a monoclonal antibody having a variable region derived from a human germline immunoglobulin sequence, and an optional constant region. In one embodiment, human monoclonal antibodies are produced by, for example, hybridomas obtained by fusing B cells obtained from transgenic or transchromosomal non-human animals (e.g., transgenic mice having a genome comprising human heavy chain transgenes and light chain transgenes) and immortal cells.

[0326] As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, produced or isolated by recombinant means, such as (a) antibodies isolated from animals (e.g., mice) transgenic or transchromosomal to human immunoglobulin genes or hybridomas prepared therefrom, (b) antibodies isolated from host cells transformed to express the antibody, such as transfectomas, (c) antibodies isolated from recombinant, combinatorial human antibody libraries, and (d) antibodies prepared, expressed, produced or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. These recombinant human antibodies include variable and constant regions that utilize specific human germline immunoglobulin sequences and are encoded by germline genes, but include subsequent rearrangements and mutations that occur during, for example, antibody maturation. As known in the art (e.g., see Lonberg (2005) Nature Biotech. 23 (9): 1117-1125), the variable region contains antigen binding domains encoded by different genes that are rearranged to form antibodies specific for foreign antigens. In addition to rearrangement, the variable region may be further modified by multiple single amino acid changes (called somatic mutations or hypermutations) to increase the affinity of the antibody for foreign antigens. The constant region will further change in response to antigen (i.e., isotype switching). Thus, the rearranged and somatically mutated nucleic acid sequences encoding light and heavy chain immunoglobulin polypeptides in response to antigen may not be identical to the original germline sequences, but will be substantially identical or similar (i.e., have at least 80% identity).

[0327] "HuMAb" refers to an antibody having a variable region in which the framework region and the CDR region in the variable region are all derived from human germline immunoglobulin sequences. In addition, if the antibody contains a constant region, the constant region is also derived from human germline immunoglobulin sequences. The antibodies described herein may include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random mutagenesis or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been transplanted to human framework sequences. The terms "human" antibody and "fully human" antibody are used synonymously.

[0328] A "humanized" antibody refers to an antibody in which some, most or all of the amino acids outside the CDR domains of a non-human antibody are replaced by corresponding amino acids derived from human immunoglobulins. In one embodiment of a humanized form of an antibody, some, most or all of the amino acids outside the CDR domains have been replaced by amino acids from human immunoglobulins, while some, most or all of the amino acids in one or more CDR regions have not changed. Small additions, deletions, insertions, substitutions or modifications of amino acids may be tolerated as long as they do not eliminate the ability of the antibody to bind a specific antigen. A "humanized" antibody retains an antigenic specificity similar to that of the original antibody.

[0329] A "chimeric antibody" refers to an antibody in which the variable region is derived from one species and the constant region is derived from another species, for example, an antibody in which the variable region is derived from a mouse antibody and the constant region is derived from a human antibody.

[0330] A "modified heavy chain constant region" refers to a heavy chain constant region comprising constant domains CH1, hinge, CH2, and CH3, wherein one or more constant domains are from different isotypes (e.g., IgG1, IgG2, IgG3, IgG4). In certain embodiments, the modified constant region comprises a human IgG2 CH1 domain and a human IgG2 hinge and a human IgG1 CH3 domain fused to a human IgG1 CH2 domain. In certain embodiments, these modified constant regions also include amino acid modifications within one or more domains relative to the wild-type amino acid sequence.

[0331] As used herein, "isotype" refers to the antibody class encoded by the heavy chain constant region genes (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibodies).

[0332] "Isotype" refers to naturally occurring variants within a particular isotype group that differ by a few amino acids (eg, see Jefferis et al. (2009) mAbs 1:1). The antibodies described herein may be of any isotype.

[0333] Unless otherwise indicated elsewhere herein, all amino acid numbering is according to the EU index of the Kabat system (Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed., US Department of Health and Human Services, NIH Publication No. 91-3242).

[0334] "An antibody that recognizes an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody that specifically binds to an antigen."

[0335] As used herein, the term "isolated antibody" is intended to refer to an antibody that is substantially free from other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to CD73 is substantially free from antibodies that specifically bind to antigens other than CD73). However, an isolated antibody that specifically binds to an epitope of CD73 may have cross-reactivity against other CD73 proteins of different species.

[0336] As used herein, an antibody that "inhibits CD73" refers to an antibody that inhibits the biological and / or enzymatic function of CD73. These functions include, for example, the ability of the antibody to inhibit CD73 enzymatic activity, such as CD73-regulated adenosine production, or reduction in cAMP production.

[0337] As used herein, an "internalizable" antibody refers to an antibody that can cross the cell membrane after binding to a cell surface antigen. Internalization includes antibody-mediated receptor (e.g., CD73) internalization. In some embodiments, the antibody is expressed as a T cell. 1 / 2 Equivalent to a rate of "internalization" into cells expressing CD73 at approximately 10 min or less.

[0338] "Effector function" refers to the interaction of an antibody Fc region with an Fc receptor or ligand, or a biochemical event derived therefrom. Exemplary "effector functions" include C1q binding, complement dependent cytotoxicity (CDC), Fc receptor binding, FcγR-mediated effector functions (e.g., ADCC and antibody-dependent cell-mediated phagocytosis (ADCP)), and downregulation of cell surface receptors (e.g., B cell receptor; BCR). These effector functions generally require the combination of an Fc region with a binding domain (e.g., an antibody variable domain).

[0339] "Fc receptor" or "FcR" is a receptor that binds to the Fc region of an immunoglobulin. FcRs that bind to IgG antibodies include receptors of the FcγR family, including allelic variants and alternatively spliced ​​forms of these receptors. The FcγR family consists of three activating receptors (FcγRI, FcγRIII, and FcγRIV in mice; FcγRIA, FcγRIIA, and FcγRIIIA in humans) and one inhibitory receptor (FcγRIIB). The various properties of human FcγRs are summarized in Table A. Most innate effector cell types co-express one or more activating FcγRs and inhibitory FcγRIIB, while natural killer (NK) cells selectively express one activating Fc receptor (FcγRIII in mice and FcγRIIIA in humans), but do not express inhibitory FcγRIIB in mice and humans. Human IgG1 binds to most human Fc receptors and is considered to bind to an activating Fc receptor type equivalent to murine IgG2a.

[0340] Table A. Properties of human FcγRs

[0341]

[0342] "Hinge", "hinge domain" or "hinge region" or "antibody hinge region" refers to the domain of the heavy chain constant region that connects the CH1 domain and the CH2 domain and includes the upper, middle and lower parts of the hinge (Roux et al., J. Immunol. 1998 161: 4083). The hinge provides different flexibility between the binding region and the effector region of the antibody and also provides a site for intermolecular disulfide bonding between the two heavy chain constant regions. The term "hinge" includes wild-type hinges and variants thereof (e.g., non-naturally occurring hinges or modified hinges). For example, the term "IgG2 hinge" includes wild-type IgG2 hinges and variants having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or up to 5, 4, 3, 2 or 1 mutations (e.g., substitutions, deletions or additions).

[0343] The term "CH1 domain" refers to the heavy chain constant region that connects the variable domain to the hinge in the heavy chain constant domain. The term "CH1 domain" includes wild-type CH1 domains and variants thereof (e.g., non-naturally occurring CH1 domains or modified CH1 domains). For example, the term "CH1 domain" includes wild-type CH1 domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or at most 5, 4, 3, 2 or 1 mutation (e.g., substitution, deletion or addition).

[0344] Exemplary CH1 domains include CH1 domains with mutations that alter the biological activity of the antibody (eg, ADCC, CDC, or half-life). Provided herein are CH1 domain modifications that affect the biological activity of an antibody.

[0345] The term "CH2 domain" refers to the heavy chain constant region connecting the hinge in the heavy chain constant domain to the CH3 domain. The term "CH2 domain" includes wild-type CH2 domains and variants thereof (e.g., non-naturally occurring CH2 domains or modified CH2 domains). For example, the term "CH2 domain" includes wild-type CH2 domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or up to 5, 4, 3, 2 or 1 mutations (e.g., substitutions, deletions or additions). Exemplary CH2 domains include CH2 domains with mutations that alter the biological activity of the antibody (e.g., ADCC, CDC or half-life).

[0346] The term "CH3 domain" refers to the heavy chain constant region that is the C-terminal end of the CH2 domain in the heavy chain constant domain. The term "CH3 domain" includes wild-type CH3 domains and variants thereof (e.g., non-naturally occurring CH3 domains or modified CH3 domains). For example, the term "CH3 domain" includes wild-type CH3 domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or up to 5, 4, 3, 2 or 1 mutations (e.g., substitutions, deletions or additions). Exemplary CH3 domains include CH3 domains with mutations that change the biological activity (e.g., ADCC, CDC or half-life) of the antibody. CH3 domain modifications that affect the biological activity of the antibody are provided herein.

[0347] "CL domain" refers to the constant domain of the light chain. The term "CL domain" includes wild-type CL domain and variants thereof.

[0348] A "native sequence Fc region" or "native sequence Fc" comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include native sequence human IgG1 Fc regions; native sequence human IgG2 Fc regions; native sequence human IgG3 Fc regions; and native sequence human IgG4 Fc regions and natural variants thereof. Native sequence Fc includes multiple isotypes of Fc (e.g., see Jefferis et al. (2009) mAbs 1:1).

[0349] The term "epitope" or "antigenic determinant" refers to a site on an antigen (e.g., CD73) to which an immunoglobulin or antibody specifically binds. Epitopes within a protein antigen can be formed from both contiguous amino acids (usually linear epitopes) or non-contiguous amino acids juxtaposed by the tertiary folding of the protein (usually conformational epitopes). Epitopes formed from contiguous amino acids are generally, but not always, retained upon exposure to denaturing solvents, while epitopes formed by tertiary folding are generally lost upon treatment with denaturing solvents. An epitope generally includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in a unique spatial conformation. Methods for determining epitope binding by a given antibody (i.e., epitope mapping) are well known in the art and include, for example, immunoblotting and immunoprecipitation analysis, in which overlapping or contiguous peptides (e.g., from CD73) are tested for reactivity with a given antibody (e.g., anti-CD73 antibody). Methods for determining the spatial conformation of epitopes include techniques in the art and those described herein, such as x-ray crystallography, 2-dimensional nuclear magnetic resonance, and HDX-MS (see, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, GE Morris, ed. (1996)).

[0350] The term "epitope mapping" refers to a process for identifying molecular determinants on an antigen that are involved in antibody-antigen recognition.

[0351] The term "binds to the same epitope" with respect to two or more antibodies means that these antibodies bind to the same stretch of amino acid residues as determined by a given method. Techniques for determining whether an antibody binds to the "same epitope on CD73" as an antibody described herein include, for example, epitope mapping methods, such as x-ray analysis of crystals of the antigen: antibody complex (which provides atomic resolution of the epitope) and hydrogen / deuterium exchange mass spectrometry (HDX-MS). Other methods monitor the binding of antibodies to antigen fragments (e.g., proteolytic fragments) or mutational variations of the antigen, where loss of binding due to modification of amino acid residues within the antigen sequence is generally considered to be indicative of epitope composition (e.g., alanine scanning mutagenesis-Cunningham and Wells (1985) Science 244:1081). In addition, computational combinatorial methods of epitope mapping may also be used. These methods rely on the ability of the antibody of interest to affinity isolate specific short peptides from combinatorial phage-displayed peptide libraries.

[0352] An antibody that "competes with another antibody for binding to a target" refers to an antibody that inhibits (partially or completely) the binding of another antibody to a target. Known competition experiments (such as those described in the Examples) can be used to determine whether two antibodies compete with each other for binding to a target, i.e., whether one antibody inhibits the binding of another antibody to a target and the degree of inhibition. In certain embodiments, an antibody competes with another antibody for binding to a target and inhibits at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% of the binding. The degree of inhibition or competition may vary depending on which antibody is a "blocking antibody" (i.e., a cold antibody that is first incubated with the target). Competition assays can be performed, for example, as described in Ed Harlow and David Lane, Cold Spring Harb Protoc; 2006; doi: 10.1101 / pdb.prot4277 or Chapter 11 of "Using Antibodies", Ed Harlow and David Lane, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999. Competing antibodies bind to the same epitope, overlapping epitopes, or adjacent epitopes (e.g., as evidenced by steric hindrance).

[0353] Other competitive binding assays include: solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614 (1986)); solid phase direct labeled assay, solid phase direct labeled sandwich analysis (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press (1988)); solid phase direct labeled RIA using 1-125 labeling (see Morel et al., Mol. Immunol. 25(1):7 (1988)); solid phase direct biotin-avidin EIA (Cheung et al., Virology 176:546 (1990)); and direct labeled RIA. (Moldenhauer et al., Scand. J. Immunol. 32:77 (1990)).

[0354] As used herein, the terms "specific binding", "selective binding", "selectively binds" and "specifically binds" refer to the binding of an antibody to an epitope on a predetermined antigen but not to other antigens. When measured by, for example, surface plasmon resonance (SPR) technology using a predetermined antigen (e.g., recombinant human CD73) as an analyte and an antibody as a ligand or Scatchard analysis of the binding of an antibody to antigen-positive cells in a 2000 surface plasmon resonance instrument, the binding of the antibody to the antigen-positive cells is about less than 10 -7 M, for example, less than about 10 -8 M, 10 -9 M or 10 -10 M or even less equilibrium dissociation constant (KD), and (ii) bind to a predetermined antigen with an affinity at least twice that of its binding to a non-specific antigen (e.g., BSA, casein) other than the predetermined antigen or a closely related antigen. Therefore, unless otherwise indicated, an antibody that "specifically binds to human CD73" refers to an antibody that binds to human CD73 with an affinity of at least 10 -7 M or less, for example, less than about 10 -8 M, 10 -9 M or 10 -10An antibody that "cross-reacts with cynomolgus monkey CD73" is an antibody that binds to soluble or cell-bound human CD73 with a KD of 10 -7 M or less, for example, less than 10 -8 M, 10 -9 M or 10 -10 In certain embodiments, antibodies that do not cross-react with CD73 from non-human species exhibit essentially undetectable binding to these proteins in standard binding assays.

[0355] As used herein, the term "Kassoc" or "Ka" is intended to refer to the association rate constant of a particular antibody-antigen interaction, while the term "Kdis" or "Kd" is intended to refer to the dissociation rate constant of a particular antibody-antigen interaction. D " refers to the equilibrium dissociation constant, which is obtained from the ratio of Kd to Ka (i.e., Kd / Ka) and is expressed in molar concentration (M). The K of an antibody can be determined using methods well established in the art. D Determine the K value of the antibody D A preferred method is by using surface plasmon resonance, preferably using a biosensor system, such as Surface plasmon resonance system or flow cytometry or Scatchard analysis.

[0356] The term "EC50" refers to the concentration of an antibody or antigen-binding portion thereof that induces a 50% maximal response, ie, a response halfway between the maximal response and baseline, in the context of an in vitro or in vivo assay using the antibody or antigen-binding fragment thereof.

[0357] The "internalization rate" of an antibody or receptor (e.g., CD73) as mediated by an antibody (e.g., an anti-CD73 antibody) can be determined by, for example, the internalized T 1 / 2 By changing the heavy chain constant region of the antibody to a modified heavy chain constant region (e.g., one containing an IgG2 hinge and an IgG2 CH1 domain), the internalization rate of the anti-CD73 antibody can be enhanced or increased by at least 10%, 30%, 50%, 75%, 2-fold, 3-fold, 5-fold or more, thereby increasing T 1 / 2 The modified heavy chain constant region may be shortened by at least 10%, 30%, 50%, 75%, 2-fold, 3-fold, 5-fold or more. For example, the modified heavy chain constant region may increase the internalization rate, such that T 1 / 2 Instead of 10 minutes, it was shortened to 5 minutes (i.e., the internalization rate was doubled or T 1 / 2 Reduced to one-half).1 / 2 " is defined as the time to reach half of the maximum internalization starting from the time the antibody is added to the cells. The maximum internalization level can be the internalization level at the plateau on a graph of internalization versus antibody concentration. The modified heavy chain constant region can increase the maximum internalization of the antibody by at least 10%, 30%, 50%, 75%, 2-fold, 3-fold, 5-fold or more. Another way to compare the internalization efficacy of different antibodies (e.g., an antibody with a modified heavy chain constant region and the same antibody without a modified heavy chain constant region) is by comparing their internalization levels at a given antibody concentration (e.g., 100 nM) or at a given time (e.g., 2 minutes, 5 minutes, 10 minutes, or 30 minutes). Comparison of internalization levels can also be performed by comparing EC50 levels of internalization.

[0358] As used herein, the term "naturally occurring" when applied to a target refers to the fact that the target can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism (including a virus) that can be isolated from a source in nature and has not been intentionally modified by man in the laboratory is naturally occurring.

[0359] "Polypeptide" refers to a chain of at least two consecutively linked amino acid residues, and the length of the chain is unlimited. One or more amino acid residues in a protein may contain modifications, such as (but not limited to) glycosylation, phosphorylation or disulfide bonds. A "protein" may contain one or more polypeptides.

[0360] As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded, and can be cDNA. "Conservative sequence modifications" of the sequences described in the SEQ ID NOs described herein are also provided, i.e., nucleotide and amino acid sequence modifications that do not eliminate the binding of antibodies encoded by the nucleotide sequence or containing the amino acid sequence to the antigen. These conservative sequence modifications include conservative nucleotide and amino acid substitutions and nucleotide and amino acid additions and deletions. For example, modifications can be introduced into the SEQ ID NOs described herein by standard techniques known in the art (e.g., site-directed mutagenesis and PCR-mediated mutagenesis). Conservative sequence modifications include conservative amino acid substitutions, in which amino acid residues are replaced with amino acid residues with similar side chains. Families of amino acid residues with similar side chains are already defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), amino acids with non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Therefore, a predicted nonessential amino acid residue in an anti-CD73 antibody is preferably replaced by another amino acid residue from the same side chain family. Methods for identifying conservative substitutions of nucleotides and amino acids that do not abolish antigen binding are well known in the art (e.g., see Brummell et al., Biochem. 32: 1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10): 879-884 (1999); and Burks et al., Proc. Natl. Acad. Sci. USA 94: 412-417 (1997)). Alternatively, in another embodiment, mutations can be randomly introduced along all or a portion of an anti-CD73 antibody coding sequence by, for example, saturation mutagenesis, and the resulting modified anti-CD73 antibodies can be screened for improved binding activity.

[0361] For nucleic acids, the term "substantial homology" means that two nucleic acids, or designated sequences thereof, are identical in at least about 80% of the nucleotides, usually at least about 90% to 95%, and more preferably at least about 98% to 99.5% of the nucleotides when optimally aligned and compared (with appropriate insertions or deletions of nucleotides). Alternatively, substantial homology exists when segments hybridize under selective hybridization conditions to the complement of a strand.

[0362] With respect to polypeptides, the term "substantial homology" means that two polypeptides or designated sequences thereof are identical in at least about 80% of their amino acids, usually at least about 90% to 95%, and more preferably at least about 98% to 99.5% when optimally aligned and compared (with appropriate insertions or deletions of nucleotides).

[0363] The % identity between two sequences varies with the number of identical positions shared by the sequences when the sequences are optimally aligned (i.e., % homology = number of identical positions / total number of positions x 100), wherein optimal alignment is determined by taking into account the number of gaps that need to be introduced to achieve optimal alignment of the two sequences and the length of each gap. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm, as described in the non-limiting examples below.

[0364] The percent identity between two nucleotide sequences can be determined using the GAP program in the GCG software package (available at http: / / www.gcg.com) using the NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70 or 80 and a length weight of 1, 2, 3, 4, 5 or 6. The percent identity between two nucleotide or amino acid sequences can also be determined using the E. Meyers and W. Miller (CABIOS, 4: 11-17 (1989)) algorithm incorporated into the ALIGN program, version 2.0, using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. In addition, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch (J. Mol. Biol. (48): 444-453 (1970)) algorithm incorporated into the GAP program in the GCG software package (available at http: / / www.gcg.com), using a Blossum 62 matrix or a PAM250 matrix and a gap weight of 16, 14, 12, 10, 8, 6 or 4 and a length weight of 1, 2, 3, 4, 5 or 6.

[0365] The nucleic acid and protein sequences described herein can further be used as a "query sequence" to perform searches against public databases, e.g., to identify related sequences. These searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed using the NBLAST program, score = 100, wordlength = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using the XBLAST program, score = 50, wordlength = 3, to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used as described in Altschul et al. (1997) Nucleic Acids Res. 25(17):3389-3402. When using BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See www.ncbi.nlm.nih.gov.

[0366] These nucleic acids may be present in intact cells, in cell lysates, or in partially purified or substantially pure form. Nucleic acids are "isolated" or "substantially pure" when purified by standard techniques to remove other cellular components or other contaminants such as other cellular nucleic acids (e.g., other parts of chromosomes) or proteins, including alkali / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other methods well known in the art. See F. Ausubel et al., eds. Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience, New York (1987).

[0367] Nucleic acids (e.g., cDNA) can be mutated according to standard techniques to provide gene sequences. For coding sequences, these mutations may affect the amino acid sequence as desired. Specifically, DNA sequences substantially homologous to or derived from natural V sequences, D sequences, J sequences, constant sequences, switch sequences, and other such sequences described herein are contemplated.

[0368] As used herein, the term "vector" is intended to refer to a nucleic acid molecule capable of transporting another nucleic acid connected thereto. One type of vector is a "plasmid", which refers to a circular double-stranded DNA loop to which other DNA segments can be connected. Another type of vector is a viral vector, in which other DNA segments can be connected to the viral genome. Certain vectors are capable of autonomous replication in host cells into which they have been introduced (e.g., bacterial vectors and episomal mammalian vectors with bacterial replication origins). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the genome of the host cell when introduced into the host cell, and thereby replicated along with the host genome. In addition, certain vectors are capable of directing the expression of genes operably connected thereto. These vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors"). In general, expression vectors used in recombinant DNA technology are usually in the form of plasmids. In this specification, "plasmid" and "vector" are used interchangeably, because plasmids are the most commonly used forms of vectors. However, the present invention also includes other forms of expression vectors that provide equivalent functions, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses).

[0369] As used herein, the term "recombinant host cell" (or simply "host cell") is intended to refer to a cell that contains a nucleic acid that is not naturally present in the cell, and may be a cell into which a recombinant expression vector has been introduced. It should be understood that these terms are intended to refer not only to a specific individual cell, but also to the progeny of such a cell. Because mutations or environmental influences may cause certain changes in subsequent generations, the progeny may not actually be the same as the parent cell, but are still included in the scope of the term "host cell" as used herein.

[0370] As used herein, the term "antigen" refers to any natural or synthetic immunogenic substance, such as a protein, peptide or hapten. The antigen may be CD73 or a fragment thereof.

[0371] "Immune response" refers to the biological response to exogenous agents in vertebrates, which protects organisms from the effects of these agents and the diseases caused by them. Immune response is mediated by the action of cells of the immune system (such as T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells or neutrophils) and soluble macromolecules (including antibodies, cytokines and complement) produced by any of these cells or the liver, which leads to selective targeting, binding, damage, destruction and / or elimination of invasive pathogens, cells or tissues infected with pathogens, cancerous cells or other abnormal cells or (in the case of autoimmunity or pathological inflammation) normal human cells or tissues from the vertebrate body. Immune response (response) or reaction (reaction) include, for example, activation or suppression of T cells (such as effector T cells or Th cells, such as CD4+ or CD8+T cells) or suppression of Treg cells.

[0372] "Immunomodulator" or "immunomodulator" refers to an agent that can participate in regulating, controlling or changing an immune response, such as a component of a signal transduction pathway. "Regulating", "regulating" or "changing" an immune response refers to any change in the activity of an immune system cell or the cell (e.g., an effector T cell). The regulation includes stimulation or suppression of the immune system, which can be manifested as an increase or decrease in the number of different cell types, an increase or decrease in the activity of these cells, or any other change that may occur in the immune system. Both inhibitory and stimulatory immunomodulators have been discovered, some of which may have enhanced function in the tumor microenvironment. Immunomodulators can be located on the surface of T cells. "Immunomodulatory target" or "immunomodulatory target" is an immunomodulator that a substance, agent, part, compound or molecule targets for binding, and its activity is changed by the binding of the substance, agent, part, compound or molecule. Immunomodulatory targets include, for example, receptors on the cell surface ("immunomodulatory receptors") and receptor ligands ("immunomodulatory ligands").

[0373] The increased ability to stimulate immune response or immune system can be derived from the enhanced agonist activity of T cell costimulatory receptors and / or the enhanced antagonist activity of inhibitory receptors. The increased ability to stimulate immune response or immune system can be reflected by the increase multiple of EC50 or maximum activity degree in the determination (such as measuring cytokine or chemokine release, cytolytic activity (directly measured on target cells or indirectly measured via detection of CD107a or granzyme) and the change of propagation) of measuring immune response. The ability to stimulate immune response or immune system activity can be enhanced by at least 10%, 30%, 50%, 75%, 2 times, 3 times, 5 times or more.

[0374] "Immunotherapy" refers to the treatment of a subject suffering from a disease or at risk of contracting a disease or suffering a recurrence of a disease by a method involving inducing, enhancing, suppressing or otherwise altering an immune response.

[0375] "Immunostimulatory therapy" or "immunostimulatory therapy" refers to therapy that increases (induces or enhances) a subject's immune response, for example to treat cancer.

[0376] "Strengthening an endogenous immune response" means increasing the effectiveness or potency of an existing immune response of a subject. This increase in effectiveness and potency can be achieved, for example, by overcoming mechanisms that suppress the endogenous host immune response, or by stimulating mechanisms that enhance the endogenous host immune response.

[0377] "T effector" ("Teff") cells refer to T cells with cytolytic activity (e.g., CD4+ and CD8+ T cells) and T helper (Th) cells, which secrete cytokines and activate and direct other immune cells, but do not include regulatory T cells (Treg cells).

[0378] As used herein, the term "linked" refers to the association of two or more molecules. The linking may be covalent or non-covalent. The linking may also be genetically linked (i.e., fused in a recombinant manner). These links may be achieved using a variety of art-recognized techniques such as chemical coupling and recombinant protein production.

[0379] As used herein, "administering" refers to the physical introduction of a composition comprising a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Preferred routes of administration of the antibodies described herein include intravenous, intraperitoneal, intramuscular, subcutaneous, spinal or other parenteral routes of administration, such as by injection or infusion. As used herein, the phrase "parenteral administration" means a mode of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intraperitoneal, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, and in vivo electroporation. Alternatively, the antibodies described herein may be administered via non-parenteral routes, such as topical, epidermal or mucosal routes of administration, such as intranasal, oral, vaginal, rectal, sublingual or topical. Administration may also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0380] As used herein, the term "T cell-mediated response" refers to a response mediated by T cells, including effector T cells (eg, CD8+ cells) and helper T cells (eg, CD4+ cells). T cell-mediated responses include, for example, T cell toxicity and proliferation.

[0381] As used herein, the term "cytotoxic T lymphocyte (CTL) response" refers to an immune response induced by cytotoxic T cells. The CTL response is mainly mediated by CD8+ T cells.

[0382] As used herein, the terms "inhibit" or "block" (e.g., in reference to inhibition / blocking of CD73 binding or activity) are used interchangeably and encompass both partial and complete inhibition / blocking.

[0383] As used herein, "cancer" refers to a broad class of diseases characterized by uncontrolled abnormal cell growth in the body. Malignant tumors or cells may form as a result of unregulated cell division, which invade adjacent tissues and may metastasize to distant parts of the body via the lymphatic system or bloodstream.

[0384] As used herein, the term "treatment" or "treating" refers to any type of intervention or procedure performed on a subject, or the administration of an active agent to a subject, to reverse, alleviate, ameliorate, inhibit or slow or prevent the progression, development, severity or recurrence of symptoms, complications, conditions or biochemical markers associated with a disease. Prevention refers to the administration to a subject who does not have the disease to prevent the disease from occurring, or if the disease occurs, to minimize its effects.

[0385] "Hematological malignancies" include lymphomas, leukemias, myelomas or lymphoid malignancies and cancers of the spleen and lymph nodes. Exemplary lymphomas include both B-cell lymphomas and T-cell lymphomas. B-cell lymphomas include both Hodgkin's lymphomas and most non-Hodgkin's lymphomas. Non-limiting examples of B-cell lymphomas include diffuse large B-cell lymphomas, follicular lymphomas, mucosa-associated lymphoid tissue lymphomas, small cell lymphocytic lymphomas (overlapping with chronic lymphocytic leukemia), mantle cell lymphomas (MCL), Burkitt's lymphomas, mediastinal large B-cell lymphomas, Waldenstrom macroglobulinemia, nodular marginal zone B-cell lymphomas, splenic marginal zone lymphomas, intravascular large B-cell lymphomas, primary effusion lymphomas, lymphomatoid granulomatosis. Non-limiting examples of T cell lymphomas include extranodal T cell lymphoma, cutaneous T cell lymphoma, anaplastic large cell lymphoma and angioimmunoblastic T cell lymphoma. Hematological malignancies also include leukemias, such as, but not limited to, secondary leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia and acute lymphoblastic leukemia. Hematological malignancies further include myeloma, such as, but not limited to, multiple myeloma and smoldering multiple myeloma. The term "hematological malignancies" encompasses other blood and / or B cell or T cell related cancers.

[0386] The term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve or at least partially achieve the desired effect. A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of the drug that, when used alone or in combination with another therapeutic agent, can promote disease regression, as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or the prevention of impairment or disability caused by the disease. A "prophylactically effective amount" or "prophylactically effective dose" of a drug is an amount of the drug that, when administered alone or in combination with another therapeutic agent to a subject at risk of contracting a disease or suffering a recurrence of the disease, inhibits the development or recurrence of the disease. The ability of a therapeutic or prophylactic agent to promote disease regression or inhibit the development or recurrence of a disease can be assessed using a variety of methods known to those skilled in the art, such as in human subjects during clinical trials, in animal model systems that are predictive of efficacy in humans, or by measuring the activity of the agent in an in vitro assay.

[0387] For example, an anticancer agent is a drug that slows cancer progression or promotes cancer regression in a subject. In a preferred embodiment, a therapeutically effective amount of a drug promotes cancer regression to the extent of eliminating cancer. "Promoting cancer regression" means that administering an effective amount of a single drug or a combination of a drug and an anticancer agent in a patient can reduce tumor growth or size, tumor necrosis, reduce the severity of at least one disease symptom, increase the frequency and duration of disease-free periods, prevent damage or disability caused by illness, or otherwise improve disease symptoms. Pharmacological effectiveness refers to the ability of a drug to promote cancer regression in a patient. Physiological safety refers to an acceptable low level of toxicity due to drug administration or other adverse physiological effects (adverse effects) at the cell, organ and / or organism level.

[0388] For example, for the treatment of tumors, a therapeutically effective amount or dose of the drug preferably inhibits cell growth or tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and still more preferably at least about 80%, relative to an untreated subject. In the best embodiment, a therapeutically effective amount or dose of the drug completely inhibits cell growth or tumor growth, i.e., preferably inhibits 100% of cell growth or tumor growth. The ability of the compound to inhibit tumor growth can be assessed using the assay described below. As another option, this property of the composition can be assessed by examining the ability of the compound to inhibit cell growth, which inhibition can be measured in vitro by assays known to those skilled in the art. In other preferred embodiments described herein, tumor regression can be observed and can last for a period of at least about 20 days, more preferably at least about 40 days, or even more preferably at least about 60 days.

[0389] The terms "patient" and "subject" refer to any human or non-human animal receiving a prophylactic or therapeutic treatment. For example, a subject with cancer can be treated using the methods and compositions described herein. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0390] "Pharmaceutically acceptable" means the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal being treated therewith.

[0391] "Pharmaceutically acceptable carrier" encompasses pharmaceutically acceptable carriers, excipients, and diluents, and means a material, composition, or vehicle involved in carrying or transporting a pharmaceutical agent from one organ or part of a subject's body to another organ or part of a subject's body, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Example

[0392] Example 1: Screening and identification of CD73 antibodies

[0393] C57 / BL6 mice were immunized with human CD73 extracellular domain recombinant protein (huCD73, SEQ ID NO: 1). The first immunization (intraperitoneal injection) was performed with an emulsion of 50 μg huCD73 protein and complete Freund's adjuvant, the second immunization (subcutaneous injection) was performed with an emulsion of 25 μg huCD73 protein and incomplete Freund's adjuvant, the third immunization (intraperitoneal injection) was performed with an emulsion of 25 μg huCD73 protein and incomplete Freund's adjuvant, and the fourth immunization (subcutaneous injection) was performed with an emulsion of 25 μg huCD73 protein and incomplete Freund's adjuvant, and finally the final booster immunization (intraperitoneal injection) was performed with 50 μg huCD73 protein. Four days after the booster, immune spleen cells were fused with SP2 / 0 cells by electrofusion to prepare hybridoma cells. The same method was used to immunize mice and prepare phage library antibodies. Primary screening was performed using ELISA and flow cytometry, and 32 hybridoma antibodies and 2 phage antibodies were obtained that bound to human and cynomolgus monkey CD73. After that, the 293T / 17 cell line expressing huCD73 (293T / 17-huCD73) was used for further enzyme activity blocking activity screening, and finally 5 clones with CD73 enzyme activity blocking function were obtained.

[0394] Example 2: Indirect ELISA to determine the binding of antibodies to CD73

[0395] The indirect ELISA method was used to determine the binding ability of each positive clone to the soluble huCD73 protein. The soluble huCD73 was coated, and the sample was diluted in multiples and incubated with the protein. Then, the HPR-labeled secondary antibody was added for incubation, and finally TMB was added for color development. After termination, the OD450 was read. Figure 1 As shown, antibodies 66F2A8D6 (SEQ ID NOs: 118 and 119), 94A12G11F2 (SEQ ID NOs: 120 and 121), 191C3A8B9 (SEQ ID NOs: 122 and 123) from three clones and two phage chimeric antibodies S1B5 (SEQ ID NOs: 13 and 14) and JB24Chi (SEQ ID NOs: 26 and 28) all bound to soluble recombinant CD73 with sub-nanomolar affinity.

[0396] Table 1. Binding ability of positive clones to soluble huCD73 protein

[0397] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> 66F2A8D6 0.1646 3.108 0.153 94A12G11F2 0.0566 2.941 0.242 191C3A8B9 0.0498 2.748 0.279 S1B5 0.0070 2.926 0.085 JB24Chi 0.1360 3.086 0.039

[0398] Example 3: Flow cytometry to measure the binding of antibodies to native CD73 on the cell surface

[0399] This method uses the 293T / 17-huCD73 cell line to determine the binding ability of 5 antibodies to natural CD73 expressed on the cell surface. After incubating the 293T / 17-huCD73 cells with the antibodies diluted in multiples, the fluorescently labeled detection antibodies were added, the fluorescence intensity values ​​were read, and the EC 50 .like Figure 2 As shown, all antibodies bound to CD73 on the cell surface with nanomolar or sub-nanomolar affinities.

[0400] Table 2. Binding ability of positive clones to natural CD73 protein

[0401] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> 66F2A8D6 0.5465 174.2 1.664 94A12G11F2 1.132 221.7 2.077 191C3A8B9 2.671 105.4 0.828 S1B5 0.6408 14.66 0.099 JB24Chi -1.467 41.98 1.135

[0402] Example 4: Evaluation of CD73 Antibodies Blocking Soluble CD73 Recombinant Protease Activity

[0403] This test evaluates the ability of CD73 antibodies to block the 5' ectonucleotidase activity of soluble huCD73 recombinant protein. Binding of the antibody blocks the activity of huCD73 recombinant protein to break down AMP into adenosine and inorganic phosphate, which competes with ATP and inhibits the ability of luciferase to emit light. Therefore, blocking antibodies attenuate light emission, and the blocking result is expressed as a decrease in RLU values. Figure 3As shown in Figure 2, 66F2A8D6, 94A12G11F2, 191C3A8B9, antibody S1B5, and JB24Chi all inhibited the activity of soluble CD73 recombinant protease. 50 All are in the nanomolar or sub-nanomolar range.

[0404] Table 3. Blocking ability of CD73 antibodies on soluble CD73 recombinant protease activity

[0405] Antibody Bottom Top <![CDATA[IC 50 (nM) <!-- 61 -->]]> 66F2A8D6 6.361 86.08 1.956 94A12G11F2 6.213 87.77 1.496 191C3A8B9 5.553 93.72 2.407 S1B5 16.180 99.47 3.259 JB24Chi 14.710 101.70 0.401

[0406] Example 5: Blockade of cell surface 5' ectonucleotidase activity by CD73 antibody

[0407] This method was based on 293T / 17-huCD73 and 293T / 17-cynoCD73 cell lines to determine the blocking ability of CD73 antibodies on cell surface 5' ectonucleotidase activity, and further confirmed the biochemical activity of the five antibodies. Figure 4 As shown in the figure, all antibodies can inhibit the 5' ectonucleotidase activity of CD73 on the cell surface. Among them, the inhibitory activity of 191C3A8B9 antibody on huCD73 is weaker than that on monkey CD73, and the inhibitory activities of the remaining antibodies on human / monkey CD73 are similar. The IC 50 Both are at 10 -11 ~10 -12 M level (Table 4).

[0408] Table 4. Blocking ability of CD73 antibodies on cell surface 5' ectonucleotidase activity

[0409]

[0410] Example 6: Reversal effect of CD73 antibody on AMP-mediated inhibition of human CD4+ T cell proliferation

[0411] The antibody blocks CD73 enzyme activity, inhibits the production of adenosine, and thus relieves the inhibition of adenosine on human CD4+T cell proliferation. This method confirms the ability of CD73 antibodies to relieve AMP-mediated CD4+T cell proliferation inhibition in vitro; CellTiter-Glo (Promega) reagent is used to detect the ability of antibodies to block CD73 enzyme activity in human CD4+T cells; and ELISA method is used to detect IFN-γ levels in cell culture supernatant.

[0412] The results showed that 66F2A8D6, 94A12G11F2, S1B5 and JB24Chi could reverse the AMP-mediated inhibition of CD4+T cell proliferation ( Figure 5 ), and EC 50 The values ​​are close, both around 10 -11~10 -12 M level (Table 5).

[0413] Table 5. Ability of CD73 antibody to reverse AMP-mediated inhibition of human CD4+ T cell proliferation

[0414] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> 66F2A8D6 29.55 69.45 0.010 94A12G11F2 32.63 73.53 0.004 S1B5 27.36 64.14 0.013 JB24Chi 26.58 62.33 0.007

[0415] Similar to the results of CD4+T cell proliferation experiments, 66F2A8D6, 94A12G11F2, S1B5 and JB24Chi all had good enzyme activity inhibition effects ( Figure 6 ), and the inhibitory capacity of each antibody was 10 -11 ~10 -12 M level (Table 6).

[0416] Table 6. Ability of antibodies to block CD73 enzymatic activity in CD4+ T cells

[0417] Antibody Bottom Top <![CDATA[IC 50 (nM)]]> 66F2A8D6 8353547 17766093 0.011 94A12G11F2 22404704 46921097 0.008 S1B5 22907851 25212940 0.048 JB24Chi 45017675 44400168 0.009

[0418] The level of IFN-γ in the cell supernatant was detected using an ELISA detection kit (Dakewe). Figure 7 As shown, 66F2A8D6, 94A12G11F2, S1B5, and JB24Chi can stimulate CD4+ T cells to release IFN-γ. 50 Both in 10 -11 ~10 -12 M level (Table 7).

[0419] Table 7. Ability of antibodies to reverse IFN-γ release from CD4+ T cells

[0420] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> 66F2A8D6 88.7 461.4 0.0101 94A12G11F2 296.6 794.6 0.0049 S1B5 156.6 576.3 0.0411 JB24Chi 169.1 512.1 0.0132

[0421] Example 7: CD73 antibody-mediated CD73 endocytosis experiment

[0422] Fab-ZAP saporin reagent (Advanced Targeting Systems) was used to detect the endocytosis mediated by 66-hIgG2 (SEQ ID NOs: 39 and 40), 94-hIgG2 (SEQ ID NOs: 51 and 53), S1B5, and JB24Chi antibodies on MDA-MB-231 cells. Figure 8 As shown in Figure 2, all four chimeric antibodies mediated CD73 endocytosis in a dose-dependent manner. The IC 50 Both in 10 -11 M level.

[0423] Table 8. Ability of antibody-mediated CD73 internalization

[0424] Antibody Bottom Top <![CDATA[IC 50 (nM)]]> 66-hIgG2 581873 3044537 0.022 94-hIgG2 649643 2809795 0.019 S1B5 896346 3084741 0.029 JB24Chi 779820 3197304 0.012

[0425] Example 8: Antibody humanization

[0426] The amino acids in the CDR and framework regions of each antibody were numbered according to the Kabat system (see Table I). The CDR transplantation method was used to humanize the two antibodies 94A12G11F and JB24Chi. The sequence homology and structural similarity of the two mouse antibodies and the human antibodies were analyzed respectively, and on this basis, the CDRs of the mouse antibodies were transplanted to a series of human antibody framework regions. Three JB24Chi humanized antibodies and three 94A12G11F humanized antibodies were obtained by preliminary ELISA screening, and were named JB24H2L1 (SEQ ID NOs:71and 73), JB24H3L2 (SEQ ID NOs:72and74), JB24H3L3 (SEQ ID NOs:72and 75), JB94H1L3 (SEQ ID NOs:81and 85), JB94H2L1 (SEQID NOs:82and 84), and JB94H3L3 (SEQ ID NOs:83and 85), respectively. The chimeric antibody composed of 94A12G11F murine antibody Fv and human IgG1 and human kappa constant regions was named JB94Chi (SEQ ID NOS:52and 53).

[0427] Example 9: Affinity determination of humanized antibodies and chimeric antibodies

[0428] Indirect ELISA to determine the affinity of humanized antibodies: Coat 96-well ELISA plates with 1ug / ml huCD73 recombinant protein, 50ul / well, wash the plates 3 times with 200ul PBST per well, block with 1% BSA for 1 hour at room temperature, and wash the plates 3 times with 200ul PBST per well. Add 100ul / well of serially diluted antibodies and incubate at room temperature for 1 hour. Wash the plates 3 times with 200ul PBST per well, add 100ul 1:5000 diluted HRP-labeled secondary antibody to each well, and incubate at room temperature for 1 hour. Wash the plates 3 times with 200ul PBST per well, add 100ul of developing buffer to each well in the dark and react at room temperature for 5 minutes. Read the OD value with 530nm excitation, 590nm emission, and 570nm cutoff. The results are as follows: Fig. 9 As shown, the affinity of the humanized antibody to the antigen was similar to that of the parent antibody (Table 9).

[0429] Table 9. Affinity of humanized antibodies to soluble huCD73

[0430] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> JB24Chi 128.1 6601 0.0273 JB24H2L1 190.9 6293 0.0248 JB24H3L2 89.06 7139 0.0208 JB24H3L3 9.8 7306 0.0228 JB94Chi 152 8474 0.0357 JB94H1L3 48.7 8288 0.0289 JB94H2L1 -141.9 7111 0.0252 JB94H3L3 320 8897 0.0292

[0431] Flow cytometry assay: 293T / 17 cells expressing huCD73 and cynoCD73 were used to evaluate the binding ability of humanized antibodies. Recombinant cells were resuspended in PBS buffer and 2x10 6 The cells were placed in a 96-well U-shaped plate, and the humanized antibodies were diluted in multiples and incubated in a 4°C refrigerator or on ice for 1 hour, then centrifuged at 4°C 1500rpm for 3 minutes, washed three times with PBS buffer, and then incubated with diluted APC-labeled goat anti-human polyclonal antibody APC Goat anti-human IgG (Biolegend) in a 4°C refrigerator or on ice for 30 minutes. Finally, the cells were washed three times with PBS as described above and placed on a MACSQuant flow cytometer for detection and analysis. GraphPad Prism software was used to generate data graphs and statistically analyze affinity data. The results are shown in Figure 2. Fig.10 As shown, the humanized antibodies of JB24Chi and 94A12G11F were compared with the parental antibodies EC 50 The values ​​are close, both around 10 -11 M to 10 -12 M level (Table 10).

[0432] Table 10. Affinity of humanized antibodies, chimeric antibodies and cell surface huCD73 and cynoCD73 antigens

[0433]

[0434] Example 10: Blocking of cell surface 5' ectonucleotidase activity by humanized antibodies

[0435] This method uses the 293T / 17-huCD73 cell line to measure CD73-inhibited AMP hydrolysis and further confirm the biochemical activity of the humanized antibody. Fig.11 As shown, the enzyme blocking activities of humanized antibodies such as JB24H3L3, JB94H1L3, and 94H1L3hIgG2 (SEQ ID NOs: 85 and 124) were all at 10 -11 ~10 -12 M level (Table 11).

[0436] Table 11. Blocking activity of humanized antibodies against cell surface 5' ectonucleotidases

[0437]

[0438] In addition, U87-MG cells were used to measure the hydrolysis of AMP catalyzed by CD73 to further confirm the biochemical activity of the humanized antibody. U87-MG cells were trypsinized and the cell density was adjusted to 4 × 10 5 / ml, add 50μl / well to a 96-well plate. Dilute the humanized antibody in multiples with MEM culture medium, add 50μl / well to the cell wells, and incubate at 37°C for 1h. Add 100μl of AMP at a concentration of 360μM to each well and incubate at 37°C for 1h. Centrifuge at 1500rpm for 3min, transfer a certain volume of culture supernatant to an opaque 96-well flat-bottom plate (Costar, 3912), and then add 2X ATP solution to make the final reaction concentration 25uM. Finally, according to the Promega instructions, add the corresponding volume of CellTiter Glo reagent in a 1:1 ratio. After equilibration at room temperature for 5 minutes, read the luminescence value on the Perkin-Elmer Envision microplate reader, and determine the cell CD73 enzyme activity by measuring the ATP level. Use GraphPad Prism software to generate data graphs and statistically analyze enzyme kinetic data. The blocking activity of JB24Chi humanized antibody and JB94Chi humanized antibody is shown in Fig.12 As shown, IC 50 All were in the sub-nanomolar range (Table 12).

[0439] Table 12. Blocking ability of humanized antibodies on 5' ectonucleotidase activity in U87-MG cells

[0440] Antibody Bottom Top <![CDATA[IC 50 (nM)]]> JB24H2L1 5795303 11954887 0.474 JB24H3L2 4703766 12052334 0.270 JB24H3L3 5596400 12116519 0.120 JB24Chi 5780677 14683906 0.053 JB94H1L3 5605512 12073719 0.314 JB94H2L1 5565211 12085615 0.214 JB94H3L3 3402301 12844937 0.456 JB94Chi 4653071 12168782 0.257

[0441] Example 11: Reversal effect of humanized antibodies on AMP-mediated inhibition of human CD4+ T cell proliferation

[0442] This method determined the ability of humanized CD73 antibodies to relieve AMP-mediated inhibition of CD4+T cell proliferation in vitro; at the same time, CellTiter-Glo (Promega) reagent was used to detect the ability of the antibody to block the CD73 enzyme activity of human CD4+T cells; and the ELISA method was used to detect the IFN-γ level in the cell culture supernatant.

[0443] The results of the humanized antibody activity of JB24Chi and JB94Chi are as follows Fig.13 As shown, the activity of JB94H1L3, 94H1L3 hIgG2 and JB94H3L3 antibodies were all above 10 -11 ~10 -12 M level (Table 13).

[0444] Table 13. Ability of humanized CD73 antibodies to reverse AMP-mediated inhibition of human CD4+ T cell proliferation

[0445] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> JB24H2L1 22.61 81.28 0.032 JB24H3L2 6.097 86.32 0.022 JB24H3L3 16.45 85.72 0.068 JB24Chi 25.18 81.45 0.003 JB94H1L3 29.9 81.79 0.002 94H1L3 hIgG2 0.558 103.5 0.012 JB94H2L1 20.33 81.28 0.032 JB94H3L3 24.47 82.01 0.003 JB94Chi 22.86 83.57 0.003

[0446] The level of IFN-γ in the cell supernatant was detected using an ELISA detection kit (Dakewe). Fig.14 As shown, except for JB24H3L3, each humanized antibody stimulated T cells to secrete IFN-γ 50 Both in 10 -11 ~10 -12 M level (Table 14).

[0447] Table 14. Ability of humanized CD73 antibodies to reverse IFN-γ release by CD4+ T cells

[0448] Antibody Bottom Top <![CDATA[EC 50 (nM)]]> JB24H2L1 380.5 875 0.002 JB24H3L2 164.6 1527 0.015 JB24H3L3 387.6 2249 0.345 JB24Chi 474.1 1897 0.006 JB94H1L3 391.1 1165 0.001 94H1L3 hIgG2 267.7 4791 0.001 JB94H2L1 380.4 875 0.002 JB94H3L3 245.9 1419 0.003 JB94Chi 267.4 1550 0.004

[0449] The level of IFN-γ in the cell supernatant was detected by ELISA kit (Dakewe). As shown in Table 14, except for JB24H3L3, the EC of each humanized antibody to stimulate T cells to secrete IFN-γ 50 Both in 10 -11 ~10 -12 M level.

[0450] Example 12: Inhibition of 5' ectonucleotidase activity of A375 xenograft tumor model by humanized antibodies

[0451] B-NDG mice (purchased from Biocytogen) were subcutaneously inoculated with A375 cells. Three days later, antibodies JB94H1L3, 94H3L3 hIgG1 (SEQ ID NOS: 91 and 85) or PBS were intraperitoneally injected (ip) at a dose of 10 mg / kg twice a week for 2 weeks. Tumors were harvested on days 1, 2, 3, and 7 after administration. Fresh tissues were placed in a small cup containing liquid nitrogen to freeze the tissues quickly, embedded in OCT, cut into 5-6 μm thickness and air-dried at room temperature. Tumor sections were fixed with 4°C precooled acetone for 20 min, dried and stored at -80°C. Tumor sections were taken out of the -80°C freezer, equilibrated at room temperature for 5-10 min, and placed in 50 mM Tris-maleic acid buffer (pH 7.4, containing 2 mM CaCl 2 and 0.25 M sucrose) at room temperature for 1 h. After 1 h, the pre-incubation buffer was discarded and 50 mM Tris-maleic acid buffer (pH 7.4, containing 5 mM MnCl 2 , 2 mM lead nitrate, 2.5% dextran T200, 2.5 mM levamisole and 400 μM AMP), incubated at 37°C for 1.5 h; rinsed three times with tap water, and the sections were washed with 1% (NH 4 ) 2S was incubated at room temperature for 1 min, followed by rapid rinsing three times with tap water; the sections were counterstained with hematoxylin and eosin, dehydrated, transparentized with xylene, and observed and photographed under a microscope. Brown indicates the presence of activated CD73, and the absence of brown indicates that the antibody has blocked the enzymatic activity of CD73. The results are shown in Figures 15 and Fig. 15B shown.

[0452] The above results show that both JB94H1L3 and 94H3L3 hIgG1 can significantly reduce the 5' exonucleotidase activity of xenograft animal model tumors, and are related to the time of administration. On the third day after drug withdrawal, the CD73 enzyme activity in the JB94H1L3-treated group dropped to the lowest point, and then recovered slightly on the seventh day. The CD73 enzyme activity in the 94H3L3 hIgG1-treated group slowly recovered from the first day after administration. In summary, both antibodies showed a good inhibitory effect on CD73 activity.

[0453] Example 13: Antitumor effect of humanized antibodies in A375 human melanoma xenograft model

[0454] A375 human melanoma cells were inoculated subcutaneously in the right flank of NCG mice, and PBMCs were resuspended in PBS and inoculated into the mice. The tumors grew to 50-100 mm 3 The tumor volume was measured twice a week using a vernier caliper to measure the long diameter and short diameter of the tumor. The volume was calculated using the formula: volume = 0.5 × long diameter × short diameter 2 The results are as follows Fig.16 As shown, JB94H1L3 exhibited significant tumor growth inhibition.

[0455] The above results were obtained using the following materials and methods:

[0456] Antibody preparation:

[0457] Preparation of hybridoma antibodies: Hybridoma cell lines were revived and cultured in DMEM medium containing 10% FBS for 7-10 days, the cell culture supernatant was collected, centrifuged at 1000 r / min for 10 min, and the supernatant was purified using Protein G HP SpinTrap (GE Healthcare). UV5nano (Millipore) microspectrophotometer was used for quantification.

[0458] Recombinant antibody expression preparation: ExpiCHO (Thermo Fisher Scientific) cells with an activity of 99% were cultured to a density of 6×10 6 / ml, centrifuge the cells at 1500rpm / min for 5min and resuspend the cells. Prepare the DNA dilution of the expression vector, ExpiFectamine CHO reagent dilution and DNA-ExpiFectamine CHO complex according to the instructions, and transfer the DNA-ExpiFectamine complex to the cell culture medium. Culture in a shaker at 37℃, 8% CO2 for 8 days. Centrifuge at 11000rpm for 10min to collect the cell supernatant. Purify the antibody using NGC chromatography system (Bio-Rad) protein purifier and rProtein G Beads 4FF pre-packed column. Quantify using UV5nano (Millipore) microspectrophotometer.

[0459] Assay 1: Indirect ELISA

[0460] 1ug / ml recombinant CD73 protein was coated on an ELISA plate (Corning) and incubated overnight at 4°C. The next day, the plate was washed 5 times with PBS buffer and blocked with 200ul / well of 2% skim milk powder; a certain dose range of CD73 antibodies was incubated at room temperature for 1h; then washed 5 times with PBST washing buffer (PBS, 0.05% Tween 20), and HRP-conjugated goat anti-mouse IgG (H+L) (Promega) was added and incubated at room temperature for 1h to detect the bound anti-CD73 antibody; the plate was washed 5 times again, and TMB (Life Technologies) was added for color development for 5 to 10min; finally, 1N HCl was added to terminate the reaction, and the OD value was measured at 450nm. Data graphs were generated and affinity data were statistically analyzed using GraphPad Prism software.

[0461] Assay 2: Flow cytometry

[0462] 100 μL of 2×10 5 Cells expressing huCD73 (293T / 17-huCD73), cynoCD73 (293T / 17-cynoCD73) or moCD73 (293T / 17-moCD73) were added to each well. The samples were diluted in a gradient manner and incubated at 4°C for 1 hour. After washing, 100 μL of APC-labeled secondary antibody was added to each well and incubated for 30 minutes. After washing, the fluorescence value was read by the machine. GraphPad Prism software was used to generate data graphs and statistically analyze affinity data.

[0463] Construction of recombinant host cell lines 293T / 17-huCD73, 293T / 17-cynoCD73, and 293T / 17-moD73: PLVX-EF1a viral vectors carrying huCD73, cynoCD73, and moCD73 genes and having puromycin resistance were used. 293T / 17 cells were transfected with transfection reagents, and the culture medium containing puromycin was replaced after about 2 days. After the cells were expanded, the limiting gradient dilution method was used to select single clones, and PE-labeled anti-huCD73, cynoCD73 and moCD73 antibodies were used to detect the surface antigens of the transfected cells. The cells were then expanded and cultured for enzyme activity determination and flow cytometry.

[0464] Assay 3: Soluble CD73 enzymatic activity blocking assay

[0465] The enzyme activity blocking function of the antibody was determined based on the soluble CD73 recombinant protein. In the presence of 500ng / ml CD73 recombinant protein, a certain dose range of CD73 antibodies was added to a 96-well flat-bottom plate and incubated at 37°C for 1 hour; then AMP and ATP with a final concentration of 180uM and 25uM were added to each well and incubated at 37°C for 30min; a certain volume of cell supernatant was transferred to a blank plate, and CellTiterr-Glo (Promega) containing luciferase was added to the above wells at a ratio of 1:1, and balanced at room temperature in the dark for 5 minutes, and finally the luminescence value was measured with an Enspire microplate reader (Perkin Elmer), and the data graph was generated and the enzyme kinetic data were statistically analyzed using GraphPad Prism software.

[0466] The percentage of enzyme inhibition was assessed as follows:

[0467] ○ATP+AMP: Maximum luciferase inhibition (100%)

[0468] ○ATP+AMP+CD73: No luciferase inhibition (0%)

[0469] The residual CD73 activity was calculated according to the following formula:

[0470]

[0471] Assay 4: Cell-based CD73 enzymatic activity blocking assay

[0472] 293T / 17-huCD73 cells were digested and centrifuged at 1500rpm for 3 minutes, then the supernatant was discarded. The cells were resuspended in serum-free DMEM medium and counted using a Biorad TC20 cell counter. The cell density was adjusted and 8,000 cells were plated in a 96-well plate at 50uL / well. 50uL of the diluted antibody solution was added to the corresponding wells and placed at 37°C and 5% CO. 2 Incubate in the incubator for 1 hour; then add 100uL of AMP solution with a final concentration of 180uM and place at 37°C and 5% CO 2 Incubate for 1 hour at 1500 rpm; then centrifuge the cell plate at 1500 rpm for 3 minutes, transfer a certain volume of culture supernatant to an opaque 96-well flat-bottom plate (Costar, 3912), and add 2X ATP solution to make the final reaction concentration 25uM. Finally, according to the Promega instructions, add the corresponding volume of CellTiter Glo reagent at 1:1, balance at room temperature for 5 minutes, read the luminescence value on the Perkin-Elmer Envision microplate reader, and determine the cell CD73 enzyme activity by measuring the ATP level. Use GraphPad Prism software to generate data graphs and statistically analyze enzyme kinetic data.

[0473] Assay 5: Fab ZAP endocytosis assay

[0474] On the first day, 2000 cells / well of MDA-MB-231 cells were plated in a 96-well flat-bottom plate at 50 μl / well and incubated at 37°C with 5% CO. 2 Incubate overnight; the next day, dilute the antibody to a certain dose range with 80nM Fab-ZAP human reagent (Advanced Targeting Systems) and incubate at room temperature for 30min to allow Fab-ZAP to bind to the antibody to be tested. Then, add 50ul / well of the antibody premix to the MDA-MB-231 cell wells at 37°C and 5% CO 2 After 3 days of incubation, the cell plate was taken out and CTG reagent (Promega) was added to lyse the cells for 2 min, and then equilibrated at room temperature for 5 min. The luminescence value was measured using Enspire microplate reader (Perkin Elmer), and the cell growth curve was calculated using GraphPad Prism software. Sequence Listing <110> Beijing Jacobs Pharmaceuticals Co., Ltd. <120> Binding molecules specific for CD73 and uses thereof <130> JABBX100 <150> PCT / CN2019 / 090366 <151> 2019-06-06 <160> 127 <170> PatentIn version 3.5 <210> 1 <211> 532 <212> PRT <213> Homo sapiens <400> 1 Trp Glu Leu Thr Ile Leu His Thr Asn Asp Val His Ser Arg Leu Glu 1 5 10 15 Gln Thr Ser Glu Asp Ser Ser Lys Cys Val Asn Ala Ser Arg Cys Met 20 25 30 Gly Gly Val Ala Arg Leu Phe Thr Lys Val Gln Gln Ile Arg Arg Ala 35 40 45 Glu Pro Asn Val Leu Leu Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr 50 55 60 Ile Trp Phe Thr Val Tyr Lys Gly Ala Glu Val Ala His Phe Met Asn 65 70 75 80 Ala Leu Arg Tyr Asp Ala Met Ala Leu Gly Asn His Glu Phe Asp Asn 85 90 95 Gly Val Glu Gly Leu Ile Glu Pro Leu Leu Lys Glu Ala Lys Phe Pro 100 105 110 Ile Leu Ser Ala Asn Ile Lys Ala Lys Gly Pro Leu Ala Ser Gln Ile 115 120 125 Ser Gly Leu Tyr Leu Pro Tyr Lys Val Leu Pro Val Gly Asp Glu Val 130 135 140 Val Gly Ile Val Gly Tyr Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn 145 150 155 160 Pro Gly Thr Asn Leu Val Phe Glu Asp Glu Ile Thr Ala Leu Gln Pro 165 170 175 Glu Val Asp Lys Leu Lys Thr Leu Asn Val Asn Lys Ile Ile Ala Leu 180 185 190 Gly His Ser Gly Phe Glu Met Asp Lys Leu Ile Ala Gln Lys Val Arg 195 200 205 Gly Val Asp Val Val Val Gly Gly His Ser Asn Thr Phe Leu Tyr Thr 210 215 220 Gly Asn Pro Pro Ser Lys Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile 225 230 235 240 Val Thr Ser Asp Asp Gly Arg Lys Val Pro Val Val Gln Ala Tyr Ala 245 250 255 Phe Gly Lys Tyr Leu Gly Tyr Leu Lys Ile Glu Phe Asp Glu Arg Gly 260 265 270 Asn Val Ile Ser Ser His Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile 275 280 285 Pro Glu Asp Pro Ser Ile Lys Ala Asp Ile Asn Lys Trp Arg Ile Lys 290 295 300 Leu Asp Asn Tyr Ser Thr Gln Glu Leu Gly Lys Thr Ile Val Tyr Leu 305 310 315 320 Asp Gly Ser Ser Gln Ser Cys Arg Phe Arg Glu Cys Asn Met Gly Asn 325 330 335 Leu Ile Cys Asp Ala Met Ile Asn Asn Asn Leu Arg His Thr Asp Glu 340 345 350 Met Phe Trp Asn His Val Ser Met Cys Ile Leu Asn Gly Gly Gly Ile 355 360 365 Arg Ser Pro Ile Asp Glu Arg Asn Asn Gly Thr Ile Thr Trp Glu Asn 370 375 380 Leu Ala Ala Val Leu Pro Phe Gly Gly Thr Phe Asp Leu Val Gln Leu 385 390 395 400 Lys Gly Ser Thr Leu Lys Lys Ala Phe Glu His Ser Val His Arg Tyr 405 410 415 Gly Gln Ser Thr Gly Glu Phe Leu Gln Val Gly Gly Ile His Val Val 420 425 430 Tyr Asp Leu Ser Arg Lys Pro Gly Asp Arg Val Val Lys Leu Asp Val 435 440 445 Leu Cys Thr Lys Cys Arg Val Pro Ser Tyr Asp Pro Leu Lys Met Asp 450 455 460 Glu Val Tyr Lys Val Ile Leu Pro Asn Phe Leu Ala Asn Gly Gly Asp 465 470 475 480 Gly Phe Gln Met Ile Lys Asp Glu Leu Leu Arg His Asp Ser Gly Asp 485 490 495 Gln Asp Ile Asn Val Val Ser Thr Tyr Ile Ser Lys Met Lys Val Ile 500 505 510 Tyr Pro Ala Val Glu Gly Arg Ile Lys Ala His His His His His His 515 520 525 His His His His 530 <210> 2 <211> 532 <212> PRT <213> Macaca fascicularis <400> 2 Trp Glu Leu Thr Ile Leu His Thr Asn Asp Val His Ser Arg Leu Glu 1 5 10 15 Gln Thr Ser Glu Asp Ser Ser Lys Cys Val Asn Ala Ser Arg Cys Met 20 25 30 Gly Gly Val Ala Arg Leu Phe Thr Lys Val Gln Gln Ile Arg Arg Ala 35 40 45 Glu Pro Asn Val Leu Leu Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr 50 55 60 Ile Trp Phe Thr Val Tyr Lys Gly Ala Glu Val Ala His Phe Met Asn 65 70 75 80 Ala Leu Arg Tyr Asp Ala Met Ala Leu Gly Asn His Glu Phe Asp Asn 85 90 95 Gly Val Glu Gly Leu Ile Glu Pro Leu Leu Lys Glu Ala Lys Phe Pro 100 105 110 Ile Leu Ser Ala Asn Ile Lys Ala Lys Gly Pro Leu Ala Ser Gln Ile 115 120 125 Ser Gly Leu Tyr Leu Pro Tyr Lys Val Leu Pro Val Gly Asp Glu Val 130 135 140 Val Gly Ile Val Gly Tyr Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn 145 150 155 160 Pro Gly Thr Asn Leu Val Phe Glu Asp Glu Ile Thr Ala Leu Gln Pro 165 170 175 Glu Val Asp Lys Leu Lys Thr Leu Asn Val Asn Lys Ile Ile Ala Leu 180 185 190 Gly His Ser Gly Phe Glu Thr Asp Lys Leu Ile Ala Gln Lys Val Arg 195 200 205 Gly Val Asp Val Val Val Gly Gly His Ser Asn Thr Phe Leu Tyr Thr 210 215 220 Gly Asn Pro Pro Ser Lys Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile 225 230 235 240 Val Thr Ser Asp Asp Gly Arg Lys Val Pro Val Val Gln Ala Tyr Ala 245 250 255 Phe Gly Lys Tyr Leu Gly Tyr Leu Lys Ile Glu Phe Asp Glu Arg Gly 260 265 270 Asn Val Ile Ser Ser His Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile 275 280 285 Pro Glu Asp Pro Ser Ile Lys Ala Asp Ile Asn Lys Trp Arg Ile Lys 290 295 300 Leu Asp Asn Tyr Ser Thr Gln Glu Leu Gly Lys Thr Ile Val Tyr Leu 305 310 315 320 Asp Gly Ser Ser Gln Ser Cys Arg Phe Arg Glu Cys Asn Met Gly Asn 325 330 335 Leu Ile Cys Asp Ala Met Ile Asn Asn Asn Leu Arg His Ala Asp Glu 340 345 350 Met Phe Trp Asn His Val Ser Met Cys Ile Leu Asn Gly Gly Gly Ile 355 360 365 Arg Ser Pro Ile Asp Glu Arg Asn Asn Gly Thr Ile Thr Trp Glu Asn 370 375 380 Leu Ala Ala Val Leu Pro Phe Gly Gly Thr Phe Asp Leu Val Gln Leu 385 390 395 400 Lys Gly Ser Thr Leu Lys Lys Ala Phe Glu His Ser Val His Arg Tyr 405 410 415 Gly Gln Ser Thr Gly Glu Phe Leu Gln Val Gly Gly Ile His Val Val 420 425 430 Tyr Asp Leu Ser Arg Lys Pro Gly Asp Arg Val Val Lys Leu Asp Val 435 440 445 Leu Cys Thr Lys Cys Arg Val Pro Ser Tyr Asp Pro Leu Lys Met Asp 450 455 460 Glu Ile Tyr Lys Val Ile Leu Pro Asn Phe Leu Ala Asn Gly Gly Asp 465 470 475 480 Gly Phe Gln Met Ile Lys Asp Glu Leu Leu Arg His Asp Ser Gly Asp 485 490 495 Gln Asp Ile Asn Val Val Ser Thr Tyr Ile Ser Lys Met Lys Val Ile 500 505 510 Tyr Pro Ala Val Glu Gly Arg Ile Lys Ala His His His His His His 515 520 525 His His His His 530 <210> 3 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 3 Glu Val Gln Leu Lys Glu Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Thr Gly Tyr 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Ser Asp Ile Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Thr Glu Asp Ser Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Arg Gly Tyr Asp Glu Thr Gly Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 4 <211> 363 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 4 gaggtgcagc tgaaggagtc tggggctgag ctggtgaagc ctggggcctc agtgaagatt 60 tcctgcaaag ctactggcta tacattcact ggctactgga tagagtgggt aaagcagagg 120 cctggacgtg gccttgagtg gattggagag attttacctg gaagtgatat tactaactac 180 aatgagaagt tcaagggcaa ggccacaatc actgcagata catcctccaa cacagcctac 240 atgcaactca gcagcctgac aactgaggac tctgccatct attactgtgc aagaaggggt 300 tacgacgaga cgggctatgc tatggactac tggggtcaag gaacctcagt cacagtctcc 360 tca 363 <210> 5 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 5 Gly Tyr Trp Ile Glu 1 5 <210> 6 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 6 Glu Ile Leu Pro Gly Ser Asp Ile Thr Asn Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 7 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 7 Arg Gly Tyr Asp Glu Thr Gly Tyr Ala Met Asp Tyr 1 5 10 <210> 8 <211> 111 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 8 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 Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30 Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile His Wing Ser Asn Leu Glu Ser Gly Ile Pro Wing 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 Val Tyr Phe Cys Gln Gln Ser Lys 85 90 95 Glu Val Pro Trp Thr Phe Gly Glu Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 9 <211> 333 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 9 gatatgtga tgacccaatc tccagcttct tggctgtgt cttagggca gaggggccacc 60 atctcctgca aggccagcca aagtgttgat tatgatggtg atagctatat gaactggtac 120 caacagaaac caggacagcc acccaactc ctcatccatg ctgcatccaa tctagaatct 180 gggatcccag ccaggtttag tggcagtggg tctgggacag acttcaccct caacatccat 240 cctgtggagg aagaggatgc tgcagtgtat ttctgtcagc aaagtaagga ggttccgtgg 300 acgttcgtgg aagggaccaa gctggaaatc aaa 333 <210> 10 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 10 Lys Ala Ser Gln Ser Val Asp Tyr Asp Gly Asp Ser Tyr Met Asn 1 5 10 15 <210> 11 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 11 Ala Ala Ser Asn Leu Glu Ser 1 5 <210> 12 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 12 Gln Gln Ser Lys Glu Val Pro Trp Thr 1 5 <210> 13 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 13 Glu Val Gln Leu Lys Glu Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Thr Gly Tyr 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Ser Asp Ile Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Thr Glu Asp Ser Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Arg Gly Tyr Asp Glu Thr Gly Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 14 <211> 218 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 14 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 Lys Ala Ser Gln Ser Val Asp Tyr Asp 20 25 30 Gly Asp Ser Tyr Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile His Ala Ala Ser Asn Leu Glu Ser Gly Ile 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 Val Tyr Phe Cys Gln Gln Ser Lys 85 90 95 Glu Val Pro Trp Thr Phe Gly Glu Gly Thr Lys Leu Glu Ile Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 145 150 155 160 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 15 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 15 Glu Val Gln Leu Lys Glu Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Thr Gly Tyr Thr Phe Thr Gly Tyr 20 25 30 Trp Ile Glu Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Leu Pro Gly Ser Asp Ile Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Ile Thr Ala Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Thr Glu Asp Ser Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Arg Gly Tyr Asp Glu Thr Gly Tyr Ala Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys 210 215 220 Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270 Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser 290 295 300 Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 16 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 16 Gln Val Gln Leu Gln Gln Ser Gly Leu Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Gly Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Lys Ala Thr Leu Thr Val Asn Lys Ser Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Val Arg Cys Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 17 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 17 caggtccaac tgcagcagtc tggacttgag ctggtgaagc ctggggctc atgagatg 60 tcctgcaagg cttctggata cacattcact gactacaa tgcactgggt gaagcagagc 120 catggaaaga gccttgagtg gattgatat attaagcctc acatggtgg tactacctac 180 aaccccgaagt tcgaggggcaa ggccacattg actgtaaaca agtctccag cacagcctac 240 atggagctcc gcagcctgac atcggaggat tctgcagtct attactgtgt aagatgcgat 300 tttctctact ggtatttcga tgtctggggc acagggacca cggtcaccgt ctcctca 357 <210> 18 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 18 Asp Tyr Asn Met His 1 5 <210> 19 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 19 Tyr Ile Lys Pro His Asn Gly Gly Thr Thr Tyr Asn Pro Lys Phe Glu 1 5 10 15 Gly <210> 20 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 20 Cys Asp Phe Leu Tyr Trp Tyr Phe Asp Val 1 5 10 <210> 21 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 21 Asp Ile Val Met Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Met Thr Cys Thr Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Gly Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu 65 70 75 80 Ala Glu Asp Ala Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Met Lys 100 105 <210> 22 <211> 324 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 22 gacattgtga tgacccagtc tccagcaatc atgtctgcat ctctagggga acgggtcacc 60 atgacctgca ctgccagctc aagtgtaagt tccagttact tgcactggta ccagcagaag 120 ccaggatcct cccccaaact ctggatttat agcacatcca acctggcttc tggagtccca 180 ggtcgcttca gtggcagtgg gtctgggacc tcttactctc tcacaatcag cagcatggag 240 gctgaagatg ctgccactta ttactgccac cagtatcatc gttccccgct cacgttcggt 300 gctgggacca agctggaaat gaaa 324 <210> 23 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 23 Thr Ala Ser Ser Ser Val Ser Ser Ser Tyr Leu His 1 5 10 <210> 24 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 24 Dear Thr Dear Asn Leu Ala Dear 1 5 <210> 25 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 25 His Gln Tyr His Arg Ser Pro Leu Thr 1 5 <210> 26 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 26 Gln Val Gln Leu Gln Gln Ser Gly Leu Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Gly Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Lys Ala Thr Leu Thr Val Asn Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Val Arg Cys Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 27 <211> 445 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 27 Gln Val Gln Leu Gln Gln Ser Gly Leu Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Gly Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Lys Ala Thr Leu Thr Val Asn Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Val Arg Cys Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Thr Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu 210 215 220 Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu 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 His Glu Asp Pro Glu Val Gln 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu 290 295 300 Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 28 <211> 215 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 28 Asp Ile Val Met Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Met Thr Cys Thr Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Gly Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu 65 70 75 80 Ala Glu Asp Ala Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Met Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 29 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 29 Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Ala Lys Pro Gly Ala 1 5 10 15 Ser Val Asn Leu Ser Cys Lys Ala Ser Gly Tyr Ala 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 Tyr Ile Asn Pro Ser Ser Gly Leu Ala Lys Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Tyr Asp Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala 115 <210> 30 <211> 354 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 30 caggtccacc tgcagcagtc tggggctgaa ctggcaaaac ctggggcctc agtgaacctg 60 tcctgcaagg cttctggcta cgcctttact agttactgga tgcactgggt aaaacagagg 120 cctggacagg gtctggaatg gattggatac attaatccta gcagtggtct tgctaagtat 180 aatcagaagt tcaaagacaa ggccacattg actacagaca aatcttccaa cacagcctac 240 atgcaactga gcagcctgac atatgacgac tctgcagtct attactgtgg aagatggtta 300 cttcggcct ggtttgctta ctggggccaa gggactctgg tcactgtctc tgca 354 <210> 31 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 31 Dear Tyr Trp Met His 1 5 <210> 32 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 32 Tyr Ile Asn Pro Ser Ser Gly Leu Ala Lys Tyr Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 33 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 33 Trp Leu Leu Ser Ala Trp Phe Ala Tyr 1 5 <210> 34 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 34 Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Ile Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Gly Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35 40 45 Tyr Arg Ala Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Asn Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Glu Phe Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 35 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 35 gacatcaaga tgacccagtc tccatcttcc atatatgcat ctctaggaga gagagtcact 60 atcacttgca aggcgagtca gggcattaat acctatttaa gctggttcca gcagaaacca 120 ggaaaatctc ctaagaccct gatctatcgt gcaaacatct tggtagatgg ggtcccatca 180 aggttcagtg gcagtggatc tgggcaagat tattctctca ccatcaacag cctggagtat 240 gaagatatgg gaatttatta ttgtctacag tatgatgagt ttccgtacac gttcggaggg 300 gggaccaagc tggaaataaa a 321 <210> 36 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 36 Lys Ala Ser Gln Gly Ile Asn Thr Tyr Leu Ser 1 5 10 <210> 37 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 37 Arg Ala Asn Ile Leu Val Asp 1 5 <210> 38 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 38 Leu Gln Tyr Asp Glu Phe Pro Tyr Thr 1 5 <210> 39 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 39 Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Ala Lys Pro Gly Ala 1 5 10 15 Ser Val Asn Leu Ser Cys Lys Ala Ser Gly Tyr Ala 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 Tyr Ile Asn Pro Ser Ser Gly Leu Ala Lys Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Tyr Asp Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys 210 215 220 Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe 225 230 235 240 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 245 250 255 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe 260 265 270 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 275 280 285 Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr 290 295 300 Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 305 310 315 320 Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 325 330 335 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 340 345 350 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 355 360 365 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 370 375 380 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser 385 390 395 400 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 405 410 415 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 420 425 430 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 40 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 40 Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Ile Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Gly Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35 40 45 Tyr Arg Ala Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Asn Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Glu Phe Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 41 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 41 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala 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 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn 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 Thr Tyr Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala 115 <210> 42 <211> 354 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 42 caggtccagc tgcagcagtc tggggctgaa ctggcaaaac ctggggcctc agtgaagctg 60 tcctgcaagg cttctggcta cacctttact agttactgga tgcactgggt aaaacagagg 120 cctggacagg gtctggaatg gattggatac attaatccta gcagtggtta tactaagtcc 180 aatcagaagt tcaaggacaa ggccacattg actgcagaca aatcctccag cacagcctac 240 atgcagctga gcagcctgac atatgaggac tctgcagtct attactgtgg aagatggtta 300 ctttcggcct ggtttgctta ctggggccaa gggactctgg tcactgtctc tgca 354 <210> 43 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 43 Ser Tyr Trp Met His 1 5 <210> 44 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 44 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 45 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 45 Trp Leu Leu Ser Ala Trp Phe Ala Tyr 1 5 <210> 46 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 46 Asp Ile Arg Met Thr Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 47 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 47 gacatcagga tgacccagtc tccatcttcc atgtatgcat ctctaggaga gagagtcact 60 atcacttgca aggcgagtca ggacattaat acctatttaa gctggttcca gcagaaacca 120 ggaaaatctc ctaagtccct gatctatcgc tcaaacatct tggtagatgg ggtcccatca 180 agattcagtg gcagtggatc tggtcaagat tattctctca ccatcagcag cctggagtat 240 gaggatatgg gaatttatta ttgtctacag tatgatgact ttccgtacac gttcggaggg 300 gggaccaagc tggaaataaa a 321 <210> 48 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 48 Lys Ala Ser Gln Asp Ile Asn Thr Tyr Leu Ser 1 5 10 <210> 49 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 49 Arg Ser Asn Ile Leu Val Asp 1 5 <210> 50 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 50 Leu Gln Tyr Asp Asp Phe Pro Tyr Thr 1 5 <210> 51 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 51 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala 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 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn 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 Thr Tyr Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys 210 215 220 Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe 225 230 235 240 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 245 250 255 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe 260 265 270 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 275 280 285 Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr 290 295 300 Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 305 310 315 320 Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 325 330 335 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 340 345 350 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 355 360 365 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 370 375 380 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser 385 390 395 400 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 405 410 415 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 420 425 430 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 52 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 52 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala 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 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn 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 Thr Tyr Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 53 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 53 Asp Ile Arg Met Thr Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 54 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 54 Gln Val Gln Leu Lys Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Ala Gly Gly Ser Thr Asn Tyr Asn Ser Ala Leu Met 50 55 60 Ser Arg Leu Ser Ile Ser Lys Asp Asn Ser Lys Ser Gln Leu Phe Leu 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Arg Glu Arg Gly Ser Ser Trp Gly Thr Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser 115 <210> 55 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 55 caggtgcagc tgaaggagtc aggacctggc ctggtggcgc cctcacagag cctgtccatc 60 acttgcactg tctctgggtt ttcattaacc aactatggtg tacactgggt tcgccagcct 120 ccaggaaagg gtctggagtg gctgggagta atatgggctg gtggaagcac aaattataat 180 tcggctctca tgtccagact gagcatcagc aaagacaact ccaagagcca acttttctta 240 aaaatgaaca gtctgcaagc tgatgacaca gccatgtact actgtgccag agagaggggt 300 agtagctggg ggactatgga ctactggggt caaggaacct cagtcactgt ctcctca 357 <210> 56 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 56 Asn Tyr Gly Val His 1 5 <210> 57 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 57 Val Ile Trp Ala Gly Gly Ser Thr Asn Tyr Asn Ser Ala Leu Met Ser 1 5 10 15 <210> 58 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 58 Glu Arg Gly Ser Ser Trp Gly Thr Met Asp Tyr 1 5 10 <210> 59 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 59 Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Arg Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr 35 40 45 Asp Thr Ser Gln Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Trp Ser Ser Asn Pro Tyr Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Met Arg 100 105 <210> 60 <211> 318 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 60 caaattgttc tcaccagtc tcagcaatc atgtctgcat ctccagggga gaaggtcacc 60 atgacctgca gtgccagctc acgtgtaagt tacatgcact ggtaccagca gaagtcaggc 120 accccccca aagatggat ttatgacaca tcccactgg cttctggagt ccctgctcgc 180 ttcagtggca gtggtctgg gacctac tctccacaa tcagcagcat ggaggctgaa 240 gatgctgcca cttattactg ccagcagtgg agtagtacc catacacgtt cggagggggg 300 accaagctgg aatgaga 318 <210> 61 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 61 Ser Ala Ser Ser Arg Val Ser Tyr Met His 1 5 10 <210> 62 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 62 Asp Thr Ser Gln Leu Ala Ser 1 5 <210> 63 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 63 Gln Gln Trp Ser Ser Asn Pro Tyr Thr 1 5 <210> 64 <211> 445 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 64 Gln Val Gln Leu Lys Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Ala Gly Gly Ser Thr Asn Tyr Asn Ser Ala Leu Met 50 55 60 Ser Arg Leu Ser Ile Ser Lys Asp Asn Ser Lys Ser Gln Leu Phe Leu 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Arg Glu Arg Gly Ser Ser Trp Gly Thr Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu 210 215 220 Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu 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 His Glu Asp Pro Glu Val Gln 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu 290 295 300 Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 65 <211> 213 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 65 Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Arg Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr 35 40 45 Asp Thr Ser Gln Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Tyr Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Met Arg Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 <210> 66 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 66 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ser Pro Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Ala Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Arg Ala Thr Leu Thr Val Asp Thr Ser Ala Ser Thr Ala 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 Ser Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 67 <211> 119 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 67 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Ala Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Arg Ala Thr Leu Thr Val Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Ser Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <210> 68 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 68 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 Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 69 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 69 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 Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Ser Ala Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 70 <211> 108 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 70 Asp Ile Val 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 Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Gly Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 71 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 71 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ser Pro Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Ala Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Arg Ala Thr Leu Thr Val Asp Thr Ser Ala Ser Thr Ala 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 Ser Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 72 <211> 449 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 72 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Asn Met His Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Lys Pro His Asn Ala Gly Thr Thr Tyr Asn Pro Lys Phe 50 55 60 Glu Gly Arg Ala Thr Leu Thr Val Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Ser Asp Phe Leu Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 73 <211> 215 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 73 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 Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 74 <211> 215 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 74 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 Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Ser Ala Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 75 <211> 215 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 75 Asp Ile Val 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 Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Asn Leu Ala Ser Gly Val Pro Gly Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln 65 70 75 80 Pro Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Tyr His Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 76 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 76 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 77 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 77 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 78 <211> 118 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 78 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys 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 Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 79 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 79 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 Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 80 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <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 Lys Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 81 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 81 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 82 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 82 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 83 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 83 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys 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 Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 84 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 84 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 Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 85 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 85 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 Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 86 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 86 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 115 120 125 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 130 135 140 Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 145 150 155 160 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr 180 185 190 Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 195 200 205 Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 210 215 220 Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 225 230 235 240 Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 245 250 255 Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 260 265 270 Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 275 280 285 Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 290 295 300 His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 305 310 315 320 Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 325 330 335 Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 340 345 350 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 355 360 365 Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 370 375 380 Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe 385 390 395 400 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 405 410 415 Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 420 425 430 Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 <210> 87 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 87 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys 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 Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 115 120 125 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 130 135 140 Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 145 150 155 160 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr 180 185 190 Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 195 200 205 Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 210 215 220 Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 225 230 235 240 Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 245 250 255 Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 260 265 270 Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 275 280 285 Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 290 295 300 His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 305 310 315 320 Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 325 330 335 Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 340 345 350 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 355 360 365 Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 370 375 380 Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe 385 390 395 400 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 405 410 415 Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 420 425 430 Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 <210> 88 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 88 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 Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110 Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly 115 120 125 Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140 Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu 145 150 155 160 Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175 Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr 180 185 190 Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205 Phe Asn Arg Asn Glu Cys 210 <210> 89 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 89 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 Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110 Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly 115 120 125 Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140 Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu 145 150 155 160 Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175 Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr 180 185 190 Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205 Phe Asn Arg Asn Glu Cys 210 <210> 90 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 90 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Ser Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 91 <211> 448 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 91 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys 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 Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Ser Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 92 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 92 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagatg 60 agctgcaagg ccagcggcta caccttcacc gactacaaca tgcactgggt gagacagagc 120 cccggcaaga gcctggagtg gatcggctac atcaagcccc acaacgccgg caccacctac 180 aaccccaagt tcgagggcag agccaccctg accgtggaca ccagcgccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgt aagaagcgac 300 ttcctgtact ggtacttcga cgtgtggggc cagggcacca ccgtgaccgt gtcctca 357 <210> 93 <211> 357 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 93 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagatg 60 agctgcaagg ccagcggcta caccttcacc gactacaaca tgcactgggt gaagcagagc 120 cacggcaaga gcctggagtg gatcggctac atcaagcccc acaacgccgg caccacctac 180 aaccccaagt tcgagggcag agccaccctg accgtggaca ccagcgccag caccgcctac 240 atggagctga gaagcctgag aagcgaggac accgccgtgt actactgcgt aagaagcgat 300 tttctctact ggtatttcga tgtctggggc cagggcacca ccgtgaccgt gtcctca 357 <210> 94 <211> 324 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 94 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca ccgccagcag cagcgtgagc agcagctacc tgcactggta ccagcagaag 120 cccggcaagg cccccaagct gctgatctac agcaccagca acctggccag cggcgtgccc 180 agcagattca gcggcagcgg cagcggcacc gacttcaccc tgaccatcag cagcctgcag 240 cccgaggact tcgccaccta ctactgccac cagtatcatc gttccccgct cacgttcggc 300 gccggcacca agctggagat caag 324 <210> 95 <211> 324 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 95 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca ccgccagcag cagcgtgagc agcagctacc tgcactggta ccagcagaag 120 cccggcagcg cccccaagct gtggatctac agcaccagca acctggccag cggcgtgccc 180 agcagattca gcggcagcgg cagcggcacc gactacaccc tgaccatcag cagcctgcag 240 cccgaggact tcgccaccta ctactgccac cagtatcatc gttccccgct cacgttcggc 300 gccggcacca agctggagat caag 324 <210> 96 <211> 324 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 96 gacatcgtga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca ccgccagcag cagcgtgagc agcagctacc tgcactggta ccagcagaag 120 cccggcagca gccccaagct gtggatctac agcaccagca acctggccag cggcgtgccc 180 ggcagattca gcggcagcgg cagcggcacc gactacaccc tgaccatcag cagcctgcag 240 cccgaggact tcgccaccta ctactgccac cagtatcatc gttccccgct cacgttcggc 300 gccggcacca agctggagat caag 324 <210> 97 <211> 1347 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 97 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagatg 60 agctgcaagg ccagcggcta caccttcacc gactacaaca tgcactgggt gagacagagc 120 cccggcaaga gcctggagtg gatcggctac atcaagcccc acaacgccgg caccacctac 180 aaccccaagt tcgagggcag agccaccctg accgtggaca ccagcgccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgt aagaagcgac 300 ttcctgtact ggtacttcga cgtgtggggc cagggcacca ccgtgaccgt gtcctcagct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct ggggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagttg 1080 accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200 gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 1260 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 1320 aagagcctct ccctgtctcc gggtaaa 1347 <210> 98 <211> 1347 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 98 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagatg 60 agctgcaagg ccagcggcta caccttcacc gactacaaca tgcactgggt gaagcagagc 120 cacggcaaga gcctggagtg gatcggctac atcaagcccc acaacgccgg caccacctac 180 aaccccaagt tcgagggcag agccaccctg accgtggaca ccagcgccag caccgcctac 240 atggagctga gaagcctgag aagcgaggac accgccgtgt actactgcgt aagaagcgat 300 tttctctact ggtatttcga tgtctggggc cagggcacca ccgtgaccgt gtcctcagct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaactcct ggggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagttg 1080 accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200 gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 1260 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 1320 aagagcctct ccctgtctcc gggtaaa 1347 <210> 99 <211> 645 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 99 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca ccgccagcag cagcgtgagc agcagctacc tgcactggta ccagcagaag 120 cccggcaagg cccccaagct gctgatctac agcaccagca acctggccag cggcgtgccc 180 agcagattca gcggcagcgg cagcggcacc gacttcaccc tgaccatcag cagcctgcag 240 cccgaggact tcgccaccta ctactgccac cagtatcatc gttccccgct cacgttcggc 300 gccggcacca agctggagat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <210> 100 <211> 645 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 100 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca ccgccagcag cagcgtgagc agcagctacc tgcactggta ccagcagaag 120 cccggcagcg cccccaagct gtggatctac agcaccagca acctggccag cggcgtgccc 180 agcagattca gcggcagcgg cagcggcacc gactacaccc tgaccatcag cagcctgcag 240 cccgaggact tcgccaccta ctactgccac cagtatcatc gttccccgct cacgttcggc 300 gccggcacca agctggagat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <210> 101 <211> 645 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 101 gacatcgtga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca ccgccagcag cagcgtgagc agcagctacc tgcactggta ccagcagaag 120 cccggcagca gccccaagct gtggatctac agcaccagca acctggccag cggcgtgccc 180 ggcagattca gcggcagcgg cagcggcacc gactacaccc tgaccatcag cagcctgcag 240 cccgaggact tcgccaccta ctactgccac cagtatcatc gttccccgct cacgttcggc 300 gccggcacca agctggagat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <210> 102 <211> 354 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 102 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaaggtg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacaggcc 120 cccggccagg gcctggagtg gatgggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agtgaccatg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg aagatggtta 300 ctttcggcct ggtttgctta ctggggccag ggcaccctgg tgaccgtgag cagc 354 <210> 103 <211> 354 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 103 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaaggtg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacagaga 120 cccggccagg gcctggagtg gatgggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agtgaccctg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg cagatggctg 300 ctgagcgcct ggttcgccta ctggggccag ggcaccctgg tgaccgtgag cagc 354 <210> 104 <211> 354 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 104 caggtgcagc tgcagcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagctg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacagaga 120 cccggccagg gcctggagtg gatcggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agccaccctg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg cagatggctg 300 ctgagcgcct ggttcgccta ctggggccag ggcaccctgg tgaccgtgag cagc 354 <210> 105 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 105 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca aggccagcca ggacatcaac acctacctga gctggttcca gcagaagccc 120 ggcaaggccc ccaagagcct gatctacaga agcaacatcc tggtggacgg cgtgcccagc 180 agattcagcg gcagcggcag cggccaggac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccatctacta ctgcctacag tatgatgact ttccgtacac gttcggccag 300 ggcaccaagc tggagatcaa g 321 <210> 106 <211> 321 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 106 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca aggccagcca ggacatcaac acctacctga gctggttcca gcagaagccc 120 ggcaagagcc ccaagagcct gatctacaga agcaacatcc tggtggacgg cgtgcccagc 180 agattcagcg gcagcggcag cggccaggac tacaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccatctacta ctgcctacag tatgatgact ttccgtacac gttcggccag 300 ggcaccaagc tggagatcaa g 321 <210> 107 <211> 1344 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 107 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaaggtg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacaggcc 120 cccggccagg gcctggagtg gatgggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agtgaccatg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg aagatggtta 300 ctttcggcct ggtttgctta ctggggccag ggcaccctgg tgaccgtgag cagcgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 720 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagttgacc 1080 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <210> 108 <211> 1344 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 108 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaaggtg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacagaga 120 cccggccagg gcctggagtg gatgggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agtgaccctg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg cagatggctg 300 ctgagcgcct ggttcgccta ctggggccag ggcaccctgg tgaccgtgag cagcgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 720 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagttgacc 1080 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <210> 109 <211> 1344 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 109 caggtgcagc tgcagcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagctg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacagaga 120 cccggccagg gcctggagtg gatcggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agccaccctg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg cagatggctg 300 ctgagcgcct ggttcgccta ctggggccag ggcaccctgg tgaccgtgag cagcgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 720 gtttctctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagttgacc 1080 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <210> 110 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 110 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca aggccagcca ggacatcaac acctacctga gctggttcca gcagaagccc 120 ggcaaggccc ccaagagcct gatctacaga agcaacatcc tggtggacgg cgtgcccagc 180 agattcagcg gcagcggcag cggccaggac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccatctacta ctgcctacag tatgatgact ttccgtacac gttcggccag 300 ggcaccaagc tggagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <210> 111 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 111 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca aggccagcca ggacatcaac acctacctga gctggttcca gcagaagccc 120 ggcaagagcc ccaagagcct gatctacaga agcaacatcc tggtggacgg cgtgcccagc 180 agattcagcg gcagcggcag cggccaggac tacaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccatctacta ctgcctacag tatgatgact ttccgtacac gttcggccag 300 ggcaccaagc tggagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <210> 112 <211> 1326 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 112 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaaggtg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacaggcc 120 cccggccagg gcctggagtg gatgggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agtgaccatg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg aagatggtta 300 ctttcggcct ggtttgctta ctggggccag ggcaccctgg tgaccgtgag cagcgccaag 360 accacccctc cttccgtgta tcctctggct ccaggatccg ccgctcagac aaactccatg 420 gtgaccctgg gttgcctggt gaagggctac ttccctgagc cagtgaccgt gacttggaac 480 tccggctctc tgtcttccgg agtgcacaca tttccagccg tgctgcagag cgacctgtac 540 acactgtcct cctccgtgac cgtgccttct tccacttggc cttccgagac cgtgacttgc 600 aacgtggccc acccagcctc ttctaccaag gtggacaaga agatcgtccc ccgggattgc 660 ggttgcaagc cttgcatttg caccgtgccc gaggtgtcct ccgtgttcat cttccctccc 720 aagcctaagg acgtgctgac catcaccctg acccccaaag tgacttgcgt ggtggtggac 780 atctctaagg acgaccccga ggtgcagttc tcttggttcg tggacgacgt ggaggtgcac 840 acagctcaga cacagccccg ggaggagcag ttcaactcca ccttccggag cgtgtccgag 900 ctgcccatca tgcaccagga ttggctgaac ggcaggagt tcaagtgccg cgtgaacagc 960 gccgctttc cagcccctat cgagagacc atctccaga ccaagggcag gcccaggct 1020 cctcaggtgt acaccatccc tcccctaag gagcagatgg ccaaggacaa ggtgtccctg 1080 acttgcatga tcaccgactt cttccccgag gattcacag tcgagtggca gtggaacggc 1140 cagccagccg agaactaca gaacacccag cccatcatgg ataccgacgg ctcttactc 1200 gtgtactcca agctgaacgt gcagaagtcc aattgggagg ccggcacac cttcacttgc 1260 tccgtgctgc acgaggact gcataccac cacaccgaga agtccctgtc ccactccc 1320 ggcaag 1326 <210> 113 <211> 1326 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 113 caggtgcagc tgcagcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagctg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacagaga 120 cccggccagg gcctggagtg gatcggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agccaccctg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg cagatggctg 300 ctgagcgcct ggttcgccta ctggggccag ggcaccctgg tgaccgtgag cagcgccaag 360 accacccctc cttccgtgta tcctctggct ccaggatccg ccgctcagac aaactccatg 420 gtgaccctgg gttgcctggt gaagggctac ttccctgagc cagtgaccgt gacttggaac 480 tccggctctc tgtcttccgg agtgcacaca tttccagccg tgctgcagag cgacctgtac 540 acactgtcct cctccgtgac cgtgccttct tccacttggc cttccgagac cgtgacttgc 600 aacgtggccc acccagcctc ttctaccaag gtggacaaga agatcgtccc ccgggattgc 660 ggttgcaagc cttgcatttg caccgtgccc gaggtgtcct ccgtgttcat cttccctccc 720 aagcctaagg acgtgctgac catcaccctg acccccaaag tgacttgcgt ggtggtggac 780 atctctaagg acgaccccga ggtgcagttc tcttggttcg tggacgacgt ggaggtgcac 840 acagctcaga cacagccccg ggaggagcag ttcaactcca ccttccggag cgtgtccgag 900 ctgcccatca tgcaccagga ttggctgaac ggcaaggagt tcaagtgccg cgtgaacagc 960 gccgcttttc cagcccctat cgagaagacc atctccaaga ccaagggcag gcccaaggct 1020 cctcaggtgt acaccatccc tccccctaag gagcagatgg ccaaggacaa ggtgtccctg 1080 acttgcatga tcaccgactt cttccccgag gacatcacag tcgagtggca gtggaacggc 1140 cagccagccg agaactacaa gaacacccag cccatcatgg ataccgacgg ctcttacttc 1200 gtgtactcca agctgaacgt gcagaagtcc aattgggagg ccggcaacac cttcacttgc 1260 tccgtgctgc acgagggact gcataaccac cacaccgaga agtccctgtc ccactctccc 1320 ggcaag 1326 <210> 114 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 114 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca aggccagcca ggacatcaac acctacctga gctggttcca gcagaagccc 120 ggcaaggccc ccaagagcct gatctacaga agcaacatcc tggtggacgg cgtgcccagc 180 agattcagcg gcagcggcag cggccaggac ttcaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccatctacta ctgcctacag tatgatgact ttccgtacac gttcggccag 300 ggcaccaagc tggagatcaa gagagccgac gccgctccta cagtgtctat cttcccccct 360 tcttccgagc agctgacctc tggaggagcc tccgtcgtgt gtttcctcaa caacttctac 420 cccaaggaca tcaacgtcaa gtggaagatc gacggctccg agaggcagaa cggcgtgctg 480 aactcttgga ccgaccagga ctccaaggac tccacctact ccatgtcctc caccctgacc 540 ctgaccaagg acgagtacga gcggcacaac tcctacactt gcgaggctac ccacaagacc 600 tctacctccc ccatcgtgaa gagcttcaac cgcaacgagt gt 642 <210> 115 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 115 gacatccaga tgacccagag ccccagcagc ctgagcgcca gcgtgggcga cagagtgacc 60 atcacctgca aggccagcca ggacatcaac acctacctga gctggttcca gcagaagcccc 120 ggcaagagcc ccaagagcct gatctacaga agcaacatcc tggtggacgg cgtgcccagc 180 agattcagcg gcagcggcag cggccaggac tacaccctga ccatcagcag cctgcagccc 240 gaggacttcg ccatctacta ctgcctacag tatgatgact ttccgtacac gttcggccag 300 ggcaccaagc tggagatcaa gagagccgac gccgctccta cagtgtctat cttcccccct 360 tcttccgagc agctgacctc tggaggagcc tccgtcgtgt gtttcctcaa caacttctac 420 ccaaggaca tcaacgtcaa gtggaagatc gacggctccg agaggcagaa cggcgtgctg 480 aactcttgga ccgaccagga ctccaaggac tccacctact ccatgtcctc caccctgacc 540 ctgaccaagg acgagtacga gcggcacaac tcctacactt gcgaggctac ccacaagacc 600 tctacctccc ccatcgtgaa gagcttcaac cgcaacgagt gt 642 <210> 116 <211> 1344 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 116 caggtgcagc tggtgcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaaggtg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacaggcc 120 cccggccagg gcctggagtg gatgggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agtgaccatg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg aagatggtta 300 ctttcggcct ggtttgctta ctggggccag ggcaccctgg tgaccgtgag cagcgctagc 360 acaaaaggac cttccgtgtt tcctctggct ccttcttcta agtctaccag cggaggaaca 420 gcagctctgg gttgtctggt gaaagattac ttcccagagc cagtgacagt gtcttggaat 480 tcaggagctc tgacatcagg agtgcataca tttccagcag tgctgcagtc ttcaggtctg 540 tattctctgt cctcagtggt gacagtgcct tcttcttctc tgggaaccca gacctacatc 600 tgtaacgtga accacaagcc ttccaacacc aaggtggata agagagtgga gcccaagtct 660 tgcgataaga cccatacttg ccctccttgt ccagctccag aatttgaagg aggaccatca 720 gtgttcctgt ttcctcctaa gcctaaggac accctgatga tctcccggac cccagaagtg 780 acttgtgtgg tggtggacgt gtctcacgaa gatcccgagg tgaagttcaa ttggtacgtg 840 gacggagtgg aagtgcataa cgctaagaca aagcctagag aggagcagta caactccaca 900 tacagagtgg tgtcagtgct gacagtgctg catcaggatt ggctgaacgg aaaggagtac 960 aagtgcaagg tgtctaacaa ggctctgcca gcttctatcg agaagaccat ctccaaggct 1020 aagggacagc ctagagaacc tcaggtgtac accctgcctc cttcccggga ggagatgaca 1080 aagaaccagg tctctctgac ttgtctggtg aagggctttt acccttccga catcgccgtg 1140 gaatgggaat ctaacggaca gccagagaac aactacaaga ccacacctcc agtgctggat 1200 tccgacggct ccttcttcct gtactccaag ctgaccgtgg ataaatctcg ttggcagcag 1260 ggaaacgtgt tctcttgtag cgtgatgcac gaagctctgc acaatcacta cacccagaag 1320 tccctgtctc tgtctccagg aaaa 1344 <210> 117 <211> 1344 <212> DNA <213> Artificial Sequence <220> <223> synthetic polynucleotide <400> 117 caggtgcagc tgcagcagag cggcgccgag gtgaagaagc ccggcgccag cgtgaagctg 60 agctgcaagg ccagcggcta caccttcacc agctactgga tgcactgggt gagacagaga 120 cccggccagg gcctggagtg gatcggctac atcaacccca gcagcggcta caccaagagc 180 aaccagaagt tcaaggacag agccaccctg accgccgaca ccagcaccag caccgcctac 240 atggagctga gcagcctgag aagcgaggac accgccgtgt actactgcgg cagatggctg 300 ctgagcgcct ggttcgccta ctggggccag ggcaccctgg tgaccgtgag cagcgctagc 360 acaaaaggac cttccgtgtt tcctctggct ccttcttcta agtctaccag cggaggaaca 420 gcagctctgg gttgtctggt gaaagattac ttcccagagc cagtgacagt gtcttggaat 480 tcaggagctc tgacatcagg agtgcataca tttccagcag tgctgcagtc ttcaggtctg 540 tattctctgt cctcagtggt gacagtgcct tcttcttctc tgggaaccca gacctacatc 600 tgtaacgtga accacaagcc ttccaacacc aaggtggata agagagtgga gcccaagtct 660 tgcgataaga cccatacttg ccctccttgt ccagctccag aatttgaagg aggaccatca 720 gtgttcctgt ttcctcctaa gcctaaggac accctgatga tctcccggac cccagaagtg 780 acttgtgtgg tggtggacgt gtctcacgaa gatcccgagg tgaagttcaa ttggtacgtg 840 gacggagtgg aagtgcataa cgctaagaca aagcctagag aggagcagta caactccaca 900 tacagagtgg tgtcagtgct gacagtgctg catcaggatt ggctgaacgg aaaggagtac 960 aagtgcaagg tgtctaacaa ggctctgcca gcttctatcg agaagaccat ctccaaggct 1020 aagggacagc ctagagaacc tcaggtgtac accctgcctc cttcccggga ggagatgaca 1080 aagaaccagg tctctctgac ttgtctggtg aagggctttt acccttccga catcgccgtg 1140 gaatgggaat ctaacggaca gccagagaac aactacaaga ccacacctcc agtgctggat 1200 tccgacggct ccttcttcct gtactccaag ctgaccgtgg ataaatctcg ttggcagcag 1260 ggaaacgtgt tctcttgtag cgtgatgcac gaagctctgc acaatcacta cacccagaag 1320 tccctgtctc tgtctccagg aaaa 1344 <210> 118 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 118 Gln Val His Leu Gln Gln Ser Gly Ala Glu Leu Ala Lys Pro Gly Ala 1 5 10 15 Ser Val Asn Leu Ser Cys Lys Ala Ser Gly Tyr Ala 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 Tyr Ile Asn Pro Ser Ser Gly Leu Ala Lys Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Tyr Asp Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 115 120 125 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 130 135 140 Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 145 150 155 160 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr 180 185 190 Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 195 200 205 Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 210 215 220 Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 225 230 235 240 Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 245 250 255 Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 260 265 270 Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 275 280 285 Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 290 295 300 His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 305 310 315 320 Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 325 330 335 Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 340 345 350 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 355 360 365 Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 370 375 380 Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe 385 390 395 400 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 405 410 415 Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 420 425 430 Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 <210> 119 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 119 Asp Ile Lys Met Thr Gln Ser Pro Ser Ser Ile Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Gly Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35 40 45 Tyr Arg Ala Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Asn Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Glu Phe Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110 Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly 115 120 125 Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140 Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu 145 150 155 160 Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175 Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr 180 185 190 Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205 Phe Asn Arg Asn Glu Cys 210 <210> 120 <211> 442 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 120 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala 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 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn 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 Thr Tyr Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 115 120 125 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 130 135 140 Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 145 150 155 160 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr 180 185 190 Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 195 200 205 Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 210 215 220 Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 225 230 235 240 Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 245 250 255 Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 260 265 270 Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 275 280 285 Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 290 295 300 His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 305 310 315 320 Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 325 330 335 Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 340 345 350 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 355 360 365 Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 370 375 380 Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe 385 390 395 400 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 405 410 415 Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 420 425 430 Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 <210> 121 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 121 Asp Ile Arg Met Thr Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Thr Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Arg Ser Asn Ile Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Tyr 65 70 75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Asp Phe Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala Ala 100 105 110 Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly 115 120 125 Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile 130 135 140 Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu 145 150 155 160 Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175 Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr 180 185 190 Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200 205 Phe Asn Arg Asn Glu Cys 210 <210> 122 <211> 443 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 122 Gln Val Gln Leu Lys Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln 1 5 10 15 Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr 20 25 30 Gly Val His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Ala Gly Gly Ser Thr Asn Tyr Asn Ser Ala Leu Met 50 55 60 Ser Arg Leu Ser Ile Ser Lys Asp Asn Ser Lys Ser Gln Leu Phe Leu 65 70 75 80 Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Arg Glu Arg Gly Ser Ser Trp Gly Thr Met Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr 115 120 125 Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu 130 135 140 Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp 145 150 155 160 Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser 180 185 190 Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser 195 200 205 Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys 210 215 220 Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro 225 230 235 240 Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr 245 250 255 Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser 260 265 270 Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg 275 280 285 Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile 290 295 300 Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn 305 310 315 320 Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys 325 330 335 Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu 340 345 350 Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe 355 360 365 Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala 370 375 380 Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr 385 390 395 400 Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly 405 410 415 Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His 420 425 430 Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 <210> 123 <211> 213 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 123 Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Arg Val Ser Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr 35 40 45 Asp Thr Ser Gln Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Tyr Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Met Arg Arg Ala Asp Ala Ala Pro 100 105 110 Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly Gly 115 120 125 Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile Asn 130 135 140 Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu Asn 145 150 155 160 Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser Ser 165 170 175 Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr Thr 180 185 190 Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser Phe 195 200 205 Asn Arg Asn Glu Cys 210 <210> 124 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 124 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys 210 215 220 Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe 225 230 235 240 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 245 250 255 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe 260 265 270 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 275 280 285 Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr 290 295 300 Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 305 310 315 320 Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 325 330 335 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 340 345 350 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 355 360 365 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 370 375 380 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser 385 390 395 400 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 405 410 415 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 420 425 430 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 125 <211> 445 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 125 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 Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Val Thr Met Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu 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 Glu Asp Pro Glu Val Gln 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440 445 <210> 126 <211> 444 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 126 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys 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 Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys 210 215 220 Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe 225 230 235 240 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 245 250 255 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe 260 265 270 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 275 280 285 Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr 290 295 300 Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 305 310 315 320 Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 325 330 335 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 340 345 350 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 355 360 365 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 370 375 380 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser 385 390 395 400 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 405 410 415 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 420 425 430 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 <210> 127 <211> 445 <212> PRT <213> Artificial Sequence <220> <223> synthetic polypeptide <400> 127 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys 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 Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Lys Ser Asn Gln Lys Phe 50 55 60 Lys Asp Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Gly Arg Trp Leu Leu Ser Ala Trp Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu 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 Glu Asp Pro Glu Val Gln 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290 295 300 Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys 325 330 335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 435 440 445

Claims

1. An isolated antibody or antigen-binding portion thereof that binds to CD73, in, The isolated antibody or antigen-binding portion thereof comprises a heavy chain variable region comprising HCDR1, HCDR2, and HCDR3 sequences; and a light chain variable region comprising LCDR1, LCDR2, and LCDR3 sequences, wherein: (a) the HCDR1 is an amino acid sequence selected from SEQ ID NO: 43; (b) the HCDR2 is an amino acid sequence selected from SEQ ID NO: 44; (c) the HCDR3 is an amino acid sequence selected from SEQ ID NO: 45; (d) the LCDR1 is an amino acid sequence selected from SEQ ID NO: 48; (e) the LCDR2 is an amino acid sequence selected from SEQ ID NO: 49; and (f) The LCDR3 is an amino acid sequence selected from SEQ ID NO:

50.

2. The isolated antibody or antigen-binding portion thereof according to claim 1, in, The isolated antibody or antigen-binding portion thereof comprises: 1) a heavy chain variable region (VH) having an amino acid sequence as shown in SEQ ID NO: 41, and a light chain variable region (VL) having an amino acid sequence as shown in SEQ ID NO: 46; 2) a heavy chain variable region (VH) having an amino acid sequence as shown in SEQ ID NO: 76, and a light chain variable region (VL) having an amino acid sequence as shown in SEQ ID NO: 79; 3) a heavy chain variable region (VH) having an amino acid sequence as shown in SEQ ID NO: 76, and a light chain variable region (VL) having an amino acid sequence as shown in SEQ ID NO: 80; 4) a heavy chain variable region (VH) having an amino acid sequence as shown in SEQ ID NO: 77, and a light chain variable region (VL) having an amino acid sequence as shown in SEQ ID NO: 79; 5) a heavy chain variable region (VH) having an amino acid sequence as shown in SEQ ID NO: 77, and a light chain variable region (VL) having an amino acid sequence as shown in SEQ ID NO: 80; 6) a heavy chain variable region (VH) having an amino acid sequence as shown in SEQ ID NO: 78, and a light chain variable region (VL) having an amino acid sequence as shown in SEQ ID NO: 79; or 7) a heavy chain variable region (VH), whose amino acid sequence is shown in SEQ ID NO: 78, and a light chain variable region (VL), whose amino acid sequence is shown in SEQ ID NO:

80.

3. The isolated antibody or antigen-binding portion thereof according to claim 1 or 2, in, The isolated antibody is IgG.

4. The isolated antibody or antigen-binding portion thereof according to claim 3, in, The isolated antibody is IgG1, IgG2 or IgG4.

5. The isolated antibody or antigen-binding portion thereof according to claim 4, in, The isolated antibody is a monoclonal antibody, a human modified antibody, a human antibody, Fv, a single chain antibody (scFv), Fab, Fab', Fab'-SH or F(ab') 2 .

6. The isolated antibody or antigen-binding portion thereof according to claim 5, in, The isolated antibody is a humanized antibody or a chimeric antibody.

7. The isolated antibody or antigen-binding portion thereof according to claim 6, in, The isolated antibody or antigen-binding portion thereof comprises: 1) a heavy chain, the amino acid sequence of which is shown in SEQ ID NO: 51, and a light chain, the amino acid sequence of which is shown in SEQ ID NO: 53; 2) a heavy chain having an amino acid sequence as shown in SEQ ID NO:52, and a light chain having an amino acid sequence as shown in SEQ ID NO:53; 3) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 81, and a light chain having an amino acid sequence as shown in SEQ ID NO: 84; 4) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 81, and a light chain having an amino acid sequence as shown in SEQ ID NO: 85; 5) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 82, and a light chain having an amino acid sequence as shown in SEQ ID NO: 84; 6) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 82, and a light chain having an amino acid sequence as shown in SEQ ID NO: 85; 7) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 83, and a light chain having an amino acid sequence as shown in SEQ ID NO: 84; 8) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 83, and a light chain having an amino acid sequence as shown in SEQ ID NO: 85; 9) a heavy chain having an amino acid sequence as shown in SEQ ID NO:90, and a light chain having an amino acid sequence as shown in SEQ ID NO:84; 10) a heavy chain having an amino acid sequence as shown in SEQ ID NO:90, and a light chain having an amino acid sequence as shown in SEQ ID NO:85; 11) a heavy chain having an amino acid sequence as shown in SEQ ID NO:91, and a light chain having an amino acid sequence as shown in SEQ ID NO:84; 12) a heavy chain having an amino acid sequence as shown in SEQ ID NO:91, and a light chain having an amino acid sequence as shown in SEQ ID NO:85; 13) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 86, and a light chain having an amino acid sequence as shown in SEQ ID NO: 88; 14) a heavy chain having an amino acid sequence as shown in SEQ ID NO:86, and a light chain having an amino acid sequence as shown in SEQ ID NO:89; 15) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 87, and a light chain having an amino acid sequence as shown in SEQ ID NO: 88; 16) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 87, and a light chain having an amino acid sequence as shown in SEQ ID NO: 89; 17) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 124, and a light chain having an amino acid sequence as shown in SEQ ID NO: 84; 18) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 124, and a light chain having an amino acid sequence as shown in SEQ ID NO: 85; 19) a heavy chain having an amino acid sequence as shown in SEQ ID NO: 125, and a light chain having an amino acid sequence as shown in SEQ ID NO: 84; or 20) a heavy chain, the amino acid sequence of which is shown in SEQ ID NO: 125, and a light chain, the amino acid sequence of which is shown in SEQ ID NO:

85.

8. An isolated antibody or antigen-binding portion thereof, in, The heavy chain comprises the amino acid sequence shown in SEQ ID NO:81, and the light chain comprises the amino acid sequence shown in SEQ ID NO:

85.

9. An isolated antibody or antigen-binding portion thereof, in, The heavy chain comprises the amino acid sequence shown in SEQ ID NO:82, and the light chain comprises the amino acid sequence shown in SEQ ID NO:

84.

10. An isolated antibody or antigen-binding portion thereof, in, The heavy chain comprises the amino acid sequence shown in SEQ ID NO:83, and the light chain comprises the amino acid sequence shown in SEQ ID NO:

85.

11. An isolated antibody or antigen-binding portion thereof, in, The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 124, and the light chain comprises the amino acid sequence shown in SEQ ID NO:

85.

12. An isolated antibody or antigen-binding portion thereof, in, The heavy chain comprises the amino acid sequence shown in SEQ ID NO: 125, and the light chain comprises the amino acid sequence shown in SEQ ID NO:

85.

13. The isolated antibody or antigen-binding portion thereof according to any one of claims 1 to 2 or 4 to 12, in, The isolated antibody or antigen-binding portion thereof is a CD73 antagonist or a 5' nucleoside enzyme antagonist of CD73.

14. The isolated antibody or antigen-binding portion thereof according to claim 13, in, The CD73 is human CD73.

15. Use of the isolated antibody or antigen binding portion thereof according to any one of claims 1 to 14 in the preparation of a medicament for reducing adenosine levels in tumor cells and / or tumor microenvironment, wherein the tumor is selected from melanoma.

16. Use of the isolated antibody or antigen binding portion thereof of any one of claims 1 to 14 in the preparation of a medicament for stimulating a T cell response in a subject suffering from a tumor, wherein the tumor is selected from melanoma.

17. Use of the isolated antibody or antigen-binding portion thereof of any one of claims 1 to 14 in the preparation of a medicament for stimulating an immune response in a subject having a tumor selected from melanoma.

18. The use according to claim 17, in, The subject has tumor cells expressing CD73 and / or a tumor microenvironment containing CD73.

19. Use of the isolated antibody or antigen-binding portion thereof of any one of claims 1 to 14 in the preparation of a medicament for inhibiting the growth of tumor cells in a subject, wherein the tumor is selected from melanoma.

20. The isolated antibody or antigen binding portion thereof of any one of claims 1 to 14 for use in reducing adenosine levels in tumor cells and / or tumor microenvironment, wherein the tumor is selected from melanoma.

21. The isolated antibody or antigen binding portion thereof of any one of claims 1 to 14 for use in stimulating a T cell response in an individual suffering from a tumor selected from melanoma.

22. The isolated antibody or antigen binding portion thereof of any one of claims 1 to 14 for use in stimulating an immune response in a subject, wherein the subject is a subject suffering from a tumor selected from melanoma.

23. The isolated antibody or antigen-binding portion thereof according to claim 22, in, The subject has tumor cells expressing CD73 and / or a tumor microenvironment containing CD73.

24. The isolated antibody or antigen binding portion thereof of any one of claims 1 to 14 for use in inhibiting the growth of tumor cells in a subject, wherein the tumor is selected from melanoma.

25. An isolated nucleic acid composition comprising: 1) comprising a first nucleic acid as shown in SEQ ID NO: 42, and a second nucleic acid as shown in SEQ ID NO: 47; 2) comprising the first nucleic acid shown in SEQ ID NO: 102, and the second nucleic acid shown in SEQ ID NO: 105; 3) containing the first nucleic acid shown in SEQ ID NO: 102, and containing the second nucleic acid shown in SEQ ID NO: 106; 4) comprising the first nucleic acid shown in SEQ ID NO: 103, and the second nucleic acid shown in SEQ ID NO: 105; 5) comprising the first nucleic acid shown in SEQ ID NO: 103, and the second nucleic acid shown in SEQ ID NO: 106; 6) comprising the first nucleic acid shown in SEQ ID NO: 104 and the second nucleic acid shown in SEQ ID NO: 105; or 7) comprising the first nucleic acid shown by SEQ ID NO: 104 and the second nucleic acid shown by SEQ ID NO:

106.

26. An expression vector composition comprising: 1) A first expression vector, in, The first expression vector contains the nucleotide sequence shown in SEQ ID NO: 42; and the second expression vector contains the nucleotide sequence shown in SEQ ID NO: 47; 2) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 102; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 105; 3) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 102; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 106; 4) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 103; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 105; 5) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 103; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 106; 6) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 104; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 105; or 7) A first expression vector, wherein the first expression vector contains the nucleotide sequence shown in SEQ ID NO: 104; and a second expression vector, wherein the second expression vector contains the nucleotide sequence shown in SEQ ID NO:

106.

27. An expression vector comprising: 1) a first nucleic acid sequence, in, The nucleotide sequence of the first nucleic acid sequence is shown in SEQ ID NO: 42; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is shown in SEQ ID NO: NO:47 shown; 2) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 102; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:105 shown; 3) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 102; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:106 shown; 4) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 103; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:105 shown; 5) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 103; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:106 shown; 6) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 104; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO: 105; or 7) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 104; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:106 shown.

28. An isolated nucleic acid composition comprising: 1) comprising a first nucleic acid as shown in SEQ ID NO: 107, and a second nucleic acid as shown in SEQ ID NO: 110; 2) containing the first nucleic acid shown in SEQ ID NO: 107, and containing the second nucleic acid shown in SEQ ID NO: 111; 3) containing the first nucleic acid shown in SEQ ID NO: 108, and containing the second nucleic acid shown in SEQ ID NO: 110; 4) comprising the first nucleic acid shown in SEQ ID NO: 108, and the second nucleic acid shown in SEQ ID NO: 111; 5) comprising the first nucleic acid shown in SEQ ID NO: 109, and the second nucleic acid shown in SEQ ID NO: 110; 6) comprising the first nucleic acid shown in SEQ ID NO: 109, and the second nucleic acid shown in SEQ ID NO: 111; 7) comprising the first nucleic acid shown in SEQ ID NO: 116, and the second nucleic acid shown in SEQ ID NO: 110; 8) comprising the first nucleic acid shown in SEQ ID NO: 116, and the second nucleic acid shown in SEQ ID NO: 111; 9) comprising the first nucleic acid shown in SEQ ID NO: 117, and the second nucleic acid shown in SEQ ID NO: 110; 10) comprising the first nucleic acid shown in SEQ ID NO: 117, and the second nucleic acid shown in SEQ ID NO: 111; 11) comprising the first nucleic acid shown in SEQ ID NO: 112, and the second nucleic acid shown in SEQ ID NO: 114; 12) comprising the first nucleic acid shown in SEQ ID NO: 112, and the second nucleic acid shown in SEQ ID NO: 115; 13) comprising the first nucleic acid shown in SEQ ID NO: 113 and the second nucleic acid shown in SEQ ID NO: 114; or 14) A first nucleic acid comprising the first nucleic acid shown in SEQ ID NO:113 and a second nucleic acid comprising the second nucleic acid shown in SEQ ID NO:

115.

29. An expression vector composition comprising: 1) A first expression vector, in, The first expression vector contains the nucleotide sequence shown in SEQ ID NO: 107; and a second expression vector, wherein the second expression vector contains the nucleotide sequence shown in SEQ ID NO: 110; 2) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 107; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111; 3) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 108; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110; 4) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 108; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111; 5) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 109; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110; 6) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 109; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111; 7) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 116; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110; 8) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 116; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111; 9) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 117; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 110; 10) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 117; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 111; 11) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 112; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 114; 12) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 112; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 115; 13) a first expression vector, wherein the first expression vector comprises the nucleotide sequence shown in SEQ ID NO: 113; and a second expression vector, wherein the second expression vector comprises the nucleotide sequence shown in SEQ ID NO: 114; or 14) A first expression vector, wherein the first expression vector contains the nucleotide sequence shown in SEQ ID NO: 113; and a second expression vector, wherein the second expression vector contains the nucleotide sequence shown in SEQ ID NO:

115.

30. An expression vector comprising: 1) a first nucleic acid sequence, in, The nucleotide sequence of the first nucleic acid sequence is shown in SEQ ID NO: 107; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is shown in SEQ ID NO: NO:110 shown; 2) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 107; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:111 shown; 3) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 108; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:110 shown; 4) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 108; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:111 shown; 5) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 109; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:110 shown; 6) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 109; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:111 shown; 7) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 116; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:110 shown; 8) a first nucleic acid sequence vector, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 116; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:111 shown; 9) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 117; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:110 shown; 10) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 117; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:111 shown; 11) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 112; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:114 shown; 12) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 112; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:115 shown; 13) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 113; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO: 114; or 14) a first nucleic acid sequence, wherein the nucleotide sequence of the first nucleic acid sequence is as shown in SEQ ID NO: 113; and a second nucleic acid sequence, wherein the nucleotide sequence of the second nucleic acid sequence is as shown in SEQ ID NO: NO:115 shown.

31. A cell comprising the expression vector according to claim 27 or 30 or the expression vector composition according to claim 26 or 29.

32. A method for preparing an isolated antibody or antigen-binding portion thereof, include: The isolated antibody or antigen binding portion thereof is expressed in the cell of claim 31 and isolated from the cell.

33. A pharmaceutical composition comprising the isolated antibody or antigen-binding portion thereof of any one of claims 1-14, and a pharmaceutically acceptable carrier.

34. A kit comprising the isolated antibody or antigen-binding portion thereof of any one of claims 1-14.

35. Use of the antibody or antigen-binding fragment thereof according to any one of claims 1 to 14, or the pharmaceutical composition according to claim 33, in the preparation of a medicament for treating a tumor, wherein the tumor is selected from melanoma.

36. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 14 or the pharmaceutical composition according to claim 33 for use in treating cancer, wherein the cancer is selected from melanoma.

Images