RU2840407C1 - Bispecific antibody - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, particularly to an antibody which specifically binds to human HLA/NY-ESO, or a binding fragment thereof, to a bispecific antibody, a pharmaceutical composition containing said antibody, as well as to methods of producing said antibodies. Also disclosed is a polynucleotide coding said antibody or a fragment thereof, a polynucleotide coding said bispecific antibody, as well as a cell and a vector containing said polynucleotides.

EFFECT: invention is effective for preventing or treating cancer with cells expressing HLA-A2/NY-ESO.

30 cl, 199 dwg, 15 tbl, 23 ex

Description

Field of technology

The present invention relates to an antibody, a bispecific antibody and the like, useful for treating tumors.

State of the art

NY-ESO-1 is a molecule that was identified in esophageal cancer by serological analysis of recombinant cDNA expression libraries (SEREX) (Non-patent Literature 1), and LAGE-1 is also referred to as "NY-ESO-2", which is a molecule identified by type difference analysis of tumor cDNA library (Non-patent Literature 2). Although the expression of such molecules is known to be localized in the testis in case of normal tissue, its mechanism remains unknown. Since the expression of NY-ESO-1 and LAGE-1 has been reported to occur in a wide variety of cancers such as melanoma, lung cancer, bladder cancer, ovarian cancer, soft tissue sarcoma and myeloma (Non-patent Literature 3), its association with cancer is also suggested. In addition, correlation with the grade of malignancy has been reported. For example, the expression level of NY-ESO-1 is higher in the metastatic lesion of melanoma than in its primary lesion (Non-Patent Literature 4), the expression levels of NY-ESO-1 and LAGE-1 are higher in late-stage urothelial carcinoma than in early-stage urothelial carcinoma (Non-Patent Literature 5), and the expression level of NY-ESO-1 is higher in high-risk myeloma with chromosomal abnormalities than in myeloma without chromosomal abnormalities (Non-Patent Literature 6). Based on such information, NY-ESO-1 and LAGE-1 have attracted attention as molecules with high cancer specificity, and a lot of research and development has been carried out to discover drugs for cancer vaccine therapy. However, there have been no approved pharmaceutical products in this regard so far.

It is known that each of the 9-mer NY-ESO peptides NY-ESO-1 and LAGE-1 contains residues 157 to 165 (i.e., SLLMWITQC), forms a complex with HLA (human leukocyte histocompatibility antigen)-A2 (i.e., HLA/NY-ESO peptide complex), and presents the complex extracellularly (Non-patent literature 7). As described above, the expression of NY-ESO-1 and LAGE-1 is cancer-specific. This indicates that the HLA/NY-ESO peptide complex is a cancer-specific therapeutic target that is selectively present on HLA-A2-positive and NY-ESO-1- or LAGE-1-positive cancer cells (Non-patent literature 8). In addition, molecules that bind to the HLA/NY-ESO peptide complex, such as TCR (Patent Document 1 and Patent Document 2) and antibodies (Patent Document 3 and Patent Document 8), have been reported.

The use of a molecule that binds to a cancer-targeting molecule is a CD3 bispecific antibody that functions based on the T cell redirection mechanism, whereby it recruits T cells to cancer cells and thus exerts an antitumor effect through cytotoxicity (Non-Patent Literature 9 and Non-Patent Literature 10). An example of a currently available CD3 bispecific antibody is Blinatumomab, which is a CD19 bispecific T cell-attracting activator (BiTE). This antibody drug is approved for the treatment of acute lymphoblastic leukemia (ALL), and clinical trials are currently underway for other types of blood cancer. However, the format of its bispecific antibody is a tandem scFv (taFv) without an Fc region, and the half-life in blood when administered to a patient is significantly shorter than that of IgG type antibodies, which are usually used as a therapeutic antibody (Non-patent literature 11).

Regarding bispecific antibodies having a heterodimeric Fc region exhibiting a half-life in blood equivalent to that of an IgG-type antibody, studies and clinical trials of CD3 bispecific antibodies with various antibody formats such as knobs and holes, CrossMAb and DuoBody® (Patent Documents 4, 5, 6 and 7) are under development, and a CD3 bispecific antibody using an antibody against an HLA/NY-ESO peptide complex has been reported (Non-Patent Document 12).

List of citations

Patent literature

Patent Literature 1: WO 2005/113595

Patent Literature 2: WO 2017/109496

Patent Literature 3: WO 2010/106431

Patent Literature 4: WO 1998/050431

Patent Literature 5: WO 2006/106905

Patent Literature 6: WO 2011/028952

Patent Literature 7: WO 2011/131746

Patent Literature 8: WO 2021/003357.

Non-patent literature

Non-patent literature 1: Proc. Natl. Acad. Sci., U.S.A., 94 (5), 1914-8, 1997

Non-patent literature 2: Int. J. Cancer, 76 (6), 903-8, 1998

Non-Patent Literature 3: Immunol. Cell Biol., 84 (3), 303-17, 2006

Non-patent literature 4: J. Surg. Res., 98 (2), 76-80, 2001

Non-Patent Literature 5: Cancer Res., 61(12), 4671-4, 2001

Non-Patent Literature 6: Blood, 105(10), 3939-44, 2005

Non-Patent Literature 7: J. Exp. Med., 187(2), 265-70, 1998

Non-patent literature 8: J. Immunol., 176 (12), 7308-16, 2006

Non-patent literature 9: Nature, 314 (6012), 628-31, 1985

Non-Patent Literature 10: Int. Rev. Immunol., 4 (2), 159-73, 1989

Non-Patent Literature 11: Drug Des. Devel. Ther., 10, 757-765, 2016

Non-Patent Literature 12: Abstracts of the 21st Annual Meeting of the Japanese Association of Cancer Immunology, O13-4.

The essence of the invention

Technical problem

The present invention provides a novel anti-HLA-A2/NY-ESO antibody that can be used as an antitumor agent, and an antitumor agent containing as an active ingredient a molecule that binds to HLA-A2/NY-ESO, containing such an antibody and the like.

Solution to the problem

The present inventors conducted concentrated studies to solve the above problem. As a result, they discovered a new anti-HLA-A2/NY-ESO antibody and a molecule that binds to HLA-A2/NY-ESO, containing such an antibody, and the like. This led to the completion of the present invention.

In particular, the present invention includes the following.

[1] An antibody that specifically binds to human HLA/NY-ESO or a binding fragment thereof, comprising:

CDRH1 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:54,

CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55,

CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56,

a light chain CDRL1 consisting of the amino acid sequence of SEQ ID NO:57, or a light chain CDRL1 consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:57, wherein amino acid 7 is W and/or amino acid 8 is K,

CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and

A light chain CDRL3 consisting of the amino acid sequence of SEQ ID NO:59, or a light chain CDRL3 consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:59, wherein amino acid 2 is A or C.

[2] The antibody according to [1], comprising CDRH1-CDRH3 and CDRL1-CDRL3 of one or more groups selected from the group consisting of groups (i)-(v) below, or a binding fragment thereof:

(i) a heavy chain CDRH1 consisting of the amino acid sequence of SEQ ID NO:54,

CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55,

CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56,

CDRL1 light chain consisting of the amino acid sequence of SEQ ID NO:57,

CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and

CDRL3 light chain consisting of the amino acid sequence of SEQ ID NO:59;

(ii) a heavy chain CDRH1 consisting of the amino acid sequence of SEQ ID NO:54,

CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55,

CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56,

a CDRL1 light chain consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:57, wherein amino acid 7 is W,

CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and

CDRL3 light chain consisting of the amino acid sequence of SEQ ID NO:59;

(iii) a heavy chain CDRH1 consisting of the amino acid sequence of SEQ ID NO:54,

CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55,

CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56,

CDRL1 light chain consisting of the amino acid sequence of SEQ ID NO:57,

CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and

a CDRL3 light chain consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:59, wherein amino acid 2 is A;

(iv) a heavy chain CDRH1 consisting of the amino acid sequence of SEQ ID NO:54,

CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55,

CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56,

CDRL1 light chain consisting of the amino acid sequence of SEQ ID NO:57,

CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and

a CDRL3 light chain consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:59, wherein amino acid 2 is S; and

(v) a heavy chain CDRH1 consisting of the amino acid sequence of SEQ ID NO:54,

CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55,

CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56,

a CDRL1 light chain consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:57, wherein amino acid 7 is W and amino acid 8 is K,

CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and

CDRL3 light chain consisting of the amino acid sequence of SEQ ID NO:59.

[3] An antibody or binding fragment thereof according to [1], comprising a heavy chain variable region consisting of an amino acid sequence having 95% or more sequence identity to the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:27, or the amino acid sequence of SEQ ID NO:38 or 39, and a light chain variable region consisting of an amino acid sequence having 95% or more amino acid sequence identity to amino acids 156-266 of the amino acid sequence of SEQ ID NO:27 or SEQ ID NO:52, or the amino acid sequence of SEQ ID NO:40.

[4] An antibody or binding fragment thereof according to [1], comprising:

(H1) a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:6,

(H2) a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:18,

(H3) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:29,

(H4) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:26,

(H5) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:27,

(H6) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:28,

(H7) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:36,

(H8) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:47,

(H9) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:48,

(H10) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:50;

(H11) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:51,

(H12) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:52,

(H13) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence SEQ ID NO:53,

(H14) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:30; or

(H15) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:156; and

(L1) a light chain variable region consisting of the amino acid sequence of SEQ ID NO:8,

(L2) a light chain variable region consisting of the amino acid sequence of SEQ ID NO:20,

(L3) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:29,

(L4) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:26,

(L5) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:27,

(L6) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:28,

(L7) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:36,

(L8) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:47,

(L9) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:48,

(L10) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:50,

(L11) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:51,

(L12) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:52,

(L13) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence SEQ ID NO:53,

(L14) a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:30; or

(L15) a light chain variable region consisting of the amino acid sequence of amino acids 161-271 of the amino acid sequence SEQ ID NO:156.

[5] An antibody or binding fragment thereof according to [4], comprising:

(H1L1) a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:6 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:8,

(H2L2) a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:18 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:20,

(H3L3) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:29, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:29,

(H4L4) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:26, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:26,

(H5L5) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:27, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:27,

(H6L6) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:28, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:28,

(H7L7) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:36, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:36,

(H8L8) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:47, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:47,

(H9L9) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:48, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:48,

(H10L10) a heavy chain variable region consisting of an amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:50, and a light chain variable region consisting of an amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:50,

(H11L11) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:51, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:51,

(H12L12) a heavy chain variable region consisting of an amino acid sequence of amino acids 21 to 140 of the amino acid sequence of SEQ ID NO:52, and a light chain variable region consisting of an amino acid sequence of amino acids 156 to 266 of the amino acid sequence of SEQ ID NO:52,

(H13L13) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:53, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:53,

(H14L14) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:30, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:30, or

(H15L14) a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:156, and a light chain variable region consisting of the amino acid sequence of amino acids 161 to 140271 of the amino acid sequence of SEQ ID NO:156.

[6] An antibody or binding fragment thereof according to any of [1] to [5], which is an scFv.

[7] An antibody or binding fragment thereof according to [6], which is

(s1) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:70,

(s2) an scFv comprising a heavy chain variable region consisting of the amino acid sequence of SEQ ID NO:18 and a light chain variable region consisting of the amino acid sequence of SEQ ID NO:20,

(s3) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:29,

(s4) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:26,

(s5) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:27,

(s6) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:28,

(s7) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:36,

(s8) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:47,

(s9) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:48,

(s10) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:50,

(s11) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:51,

(s12) scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:52,

(s13) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:53,

(s14) an scFv consisting of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:30, or

(s15) scFv consisting of the amino acid sequence of amino acids 21-271 of the amino acid sequence of SEQ ID NO:156.

[8] A polynucleotide encoding an antibody or binding fragment thereof according to any one of [1]-[7].

[9] A vector containing the polynucleotide according to [8].

[10] A host cell containing a polynucleotide according to [8] or a vector according to [9].

[11] A method for producing an antibody that specifically binds to human HLA/NY-ESO or a binding fragment thereof, comprising: (i) a step of culturing a host cell according to [10]; and (ii) a step of purifying the antibody or a binding fragment thereof from the culture product obtained in step (i).

[12] An antibody that specifically binds to human HLA/NY-ESO or a binding fragment thereof, obtained by the method of [11].

[13] An antibody that has the properties of (i) or (ii) below and binds to HLA-A2/NY-ESO or a binding fragment thereof:

(i) binding to a site on HLA-A2/NY-ESO recognized by an antibody or binding fragment thereof according to [7]; or

(ii) binding of human HLA-A2/NY-ESO in competition with the antibody or its binding fragment according to [7].

[14] A pharmaceutical composition comprising, as an active ingredient, an antibody or a binding fragment thereof according to any one of [1]-[7], [12] and [13], a polynucleotide according to [8], a vector according to [9], or a cell according to [10].

[15] A molecule that specifically binds to human HLA/NY-ESO, comprising an antibody or binding fragment thereof according to any one of [1]-[7], [12] and [13].

[16] The molecule according to [15], which is a multispecific antibody.

[17] The molecule according to [15], which is a bispecific antibody.

[18] A molecule according to any of [15]-[17], comprising an antibody that specifically binds to CD3, or a binding fragment thereof.

[19] The molecule according to [18], wherein the antibody that specifically binds to CD3, or a binding fragment thereof, comprises:

(CCH1) CDRH1 heavy chain consisting of the amino acid sequence of SEQ ID NO:141;

(CCH2) a heavy chain CDRH2 consisting of the amino acid sequence of SEQ ID NO:142, or a heavy chain CDRH2 consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:142, wherein amino acid 3 is N or S;

(CCH3) CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:143;

(CCL1) CDRL1 light chain consisting of the amino acid sequence of SEQ ID NO:144;

(CCL2) a light chain CDRL2 consisting of the amino acid sequence of SEQ ID NO:145, or a light chain CDRL2 consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:145, wherein amino acid 2 is N; and

(CCL3) CDRL3 light chain consisting of the amino acid sequence of SEQ ID NO:146.

[20] The molecule according to [19], wherein the antibody that specifically binds to CD3, or a binding fragment thereof, comprises:

(CH1) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:136,

(CH2) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:137,

(CH3) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:147,

(CH4) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:138,

(CH5) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence SEQ ID NO:139,

(CH6) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence SEQ ID NO:140,

(CH7) a heavy chain variable region consisting of the amino acid sequence of amino acids 272-389 of the amino acid sequence SEQ ID NO:155,

(CH8) a heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:156; or

(CH9) a heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:157; and

(CL1) a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:136,

(CL2) a light chain variable region consisting of the amino acid sequence of amino acids 135-241 of the amino acid sequence SEQ ID NO:137,

(CL3) a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:147,

(CL4) a light chain variable region consisting of the amino acid sequence of amino acids 135-241 of the amino acid sequence SEQ ID NO:138,

(CL5) a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:139,

(CL6) a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:140,

(CL7) a light chain variable region consisting of the amino acid sequence of amino acids 405-511 of the amino acid sequence SEQ ID NO:155,

(CL8) a light chain variable region consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence of SEQ ID NO:156, or

(CL9) a light chain variable region consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence SEQ ID NO:157.

[21] The molecule according to [20], wherein the antibody that specifically binds to CD3, or a binding fragment thereof, comprises:

(CH1CL1) a heavy chain variable region consisting of an amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:136, and a light chain variable region consisting of an amino acid sequence of amino acids 135-243 of the amino acid sequence of SEQ ID NO:136;

(CH2CL2) a heavy chain variable region consisting of an amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:137, and a light chain variable region consisting of an amino acid sequence of amino acids 135-241 of the amino acid sequence of SEQ ID NO:137;

(CH3CL3) a heavy chain variable region consisting of an amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:147, and a light chain variable region consisting of an amino acid sequence of amino acids 135-243 of the amino acid sequence of SEQ ID NO:147;

(CH4CL4) a heavy chain variable region consisting of an amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:138, and a light chain variable region consisting of an amino acid sequence of amino acids 135-241 of the amino acid sequence of SEQ ID NO:138;

(CH5CL5) a heavy chain variable region consisting of an amino acid sequence of amino acids 2 to 119 of the amino acid sequence of SEQ ID NO:139, and a light chain variable region consisting of an amino acid sequence of amino acids 135 to 243 of the amino acid sequence of SEQ ID NO:139;

(CH6CL6) a heavy chain variable region consisting of an amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:140, and a light chain variable region consisting of an amino acid sequence of amino acids 135-243 of the amino acid sequence of SEQ ID NO:140;

(CH7CL7) a heavy chain variable region consisting of an amino acid sequence of amino acids 272-389 of the amino acid sequence of SEQ ID NO:155, and a light chain variable region consisting of an amino acid sequence of amino acids 272-389 of the amino acid sequence of SEQ ID NO:155;

(CH8CL8) a heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:156, and a light chain variable region consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence of SEQ ID NO:156; or

(CH9CL9) a heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:157, and a light chain variable region consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence of SEQ ID NO:157.

[22] The molecule according to any of [18]-[21], wherein the antibody that specifically binds to CD3, or a binding fragment thereof, is an scFv.

[23] The molecule according to [22], wherein scFv comprises:

(CS1) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:136;

(CS2) an scFv consisting of the amino acid sequence of amino acids 2-241 of the amino acid sequence of SEQ ID NO:137;

(CS3) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:147;

(CS4) an scFv consisting of the amino acid sequence of amino acids 2-241 of the amino acid sequence of SEQ ID NO:138;

(CS5) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:139;

(CS6) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:140;

(CS7) an scFv consisting of the amino acid sequence of amino acids 272-511 of the amino acid sequence of SEQ ID NO:155;

(CS8) an scFv consisting of the amino acid sequence of amino acids 277-516 of the amino acid sequence of SEQ ID NO:156; or

(CS9) scFv consisting of the amino acid sequence of amino acids 277-516 of the amino acid sequence of SEQ ID NO:157.

[24] A molecule according to any one of [18] to [23], comprising: a first polypeptide comprising an scFv that specifically binds to human HLA/NY-ESO, an scFv that specifically binds to CD3, and an Fc region (i) in this order from the N terminus to the C terminus; and a second polypeptide comprising an Fc region (ii), wherein the first polypeptide is preferably linked to a second polypeptide in the Fc region (i) and the Fc region (ii).

[25] The molecule according to [24], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [1], which is an scFv.

[26] The molecule according to [24], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [2], which is an scFv.

[27] The molecule according to [24], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [3], which is an scFv.

[28] The molecule according to [24], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [4], which is an scFv.

[29] The molecule according to [24], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [5], which is an scFv.

[30] The molecule according to any one of [19]-[24], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [18], which is an scFv.

[31] The molecule according to any one of [19]-[24], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [19], which is an scFv.

[32] The molecule according to any one of [19]-[24], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [20] or [21], which is an scFv.

[33] The molecule according to any one of [19]-[24], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [23], which is an scFv.

[34] The molecule according to any one of [24] to [33], comprising an amino acid sequence selected from the group consisting of an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:88, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:89, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:90, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:91, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:92, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:93, an amino acid sequence of amino acids 21-511 amino acid sequence of SEQ ID NO:94, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:95, the amino acid sequence of amino acids 21-511 of the amino acid sequence shown in SEQ ID NO:96, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:86, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:149, and the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:150.

[35] A molecule according to any one of [24]-[33], which comprises an amino acid sequence selected from the group consisting of an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:155, an amino acid sequence of amino acids 21-516 of the amino acid sequence of SEQ ID NO:156, and an amino acid sequence of amino acids 21-516 of the amino acid sequence of SEQ ID NO:157.

[36] The molecule of any one of [24]-[34], wherein the first polypeptide comprises an amino acid sequence of amino acids 529-745 of the amino acid sequence of SEQ ID NO:85, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 86, 149 or 150.

[37] The molecule according to any one of [24]-[33] and [35], wherein the first polypeptide comprises the amino acid sequence of amino acids 529-745 of the amino acid sequence of SEQ ID NO:155, the amino acid sequence of amino acids 534-750 of the amino acid sequence of SEQ ID NO:156, or the amino acid sequence of amino acids 534-750 of the amino acid sequence of SEQ ID NO:157.

[38] The molecule of [34] or [36], wherein the first polypeptide consists of an amino acid sequence selected from the group consisting of an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:88, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:89, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:90, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:91, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:92, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:93, an amino acid sequence of amino acids 20-745 amino acid sequence of SEQ ID NO:94, the amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:95, the amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:96, the amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:86, the amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:149, and the amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:150.

[39] The molecule according to [35] or [37], wherein the first polypeptide consists of an amino acid sequence selected from the group consisting of an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:155, an amino acid sequence of amino acids 20-750 of the amino acid sequence of SEQ ID NO:156, and an amino acid sequence of amino acids 20-750 of the amino acid sequence of SEQ ID NO:157.

[40] The molecule of any one of [24]-[39], wherein the second polypeptide comprises an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84.

[41] A molecule according to any one of [18]-[23], comprising a first polypeptide, a second polypeptide and a third polypeptide, wherein

the first polypeptide comprises an scFv that specifically binds to human HLA/NY-ESO, an scFv that specifically binds to CD3, and an Fc region (i) in that order from N terminus to C terminus,

the second polypeptide comprises an immunoglobulin heavy chain containing an Fc region (ii), and

the third polypeptide comprises an immunoglobulin light chain; and

the second polypeptide is preferably associated with the third polypeptide, and

the first polypeptide is preferably associated with the second polypeptide in the Fc regions.

[42] The molecule according to [41], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [1], which is an scFv.

[43] The molecule according to [41], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [2], which is an scFv.

[44] The molecule according to [41], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [3], which is an scFv.

[45] The molecule according to [41], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [4], which is an scFv.

[46] The molecule according to [41], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [5], which is an scFv.

[47] The molecule according to any one of [42]-[46], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [18], which is an scFv.

[48] The molecule according to any one of [42]-[46], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [19], which is an scFv.

[49] The molecule according to any of [42]-[46], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [20] or [21], which is an scFv.

[50] The molecule according to any one of [42]-[46], wherein the scFv that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [23], which is an scFv.

[51] The molecule of any one of [41]-[50], wherein the second polypeptide comprises an amino acid sequence of amino acids 20-242 of the amino acid sequence of SEQ ID NO:99.

[52] The molecule of any one of [41]-[51], wherein the third polypeptide comprises the amino acid sequence shown in SEQ ID NO:100.

[53] The molecule of any one of paragraphs [41]-[52], wherein the first polypeptide comprises an amino acid sequence selected from the group consisting of an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:88, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:89, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:90, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:91, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:92, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:93, an amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:94, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:95, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:96, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:86, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:149, and the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:150.

[54] The molecule of any one of [41]-[53], wherein the first polypeptide consists of an amino acid sequence selected from the group consisting of an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:88, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:89, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:90, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:91, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:92, an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:93, an amino acid sequence of amino acids 20-745 of the amino acid sequence SEQ ID NO:94, the amino acid sequence of amino acids 20-745 of the amino acid sequence SEQ ID NO:95, the amino acid sequence of amino acids 20-745 of the amino acid sequence SEQ ID NO:96, the amino acid sequence of amino acids 20-745 of the amino acid sequence SEQ ID NO:86, the amino acid sequence of amino acids 20-745 of the amino acid sequence SEQ ID NO:149, and the amino acid sequence of amino acids 20-745 of the amino acid sequence SEQ ID NO:150.

[55] A molecule according to any one of [18] to [23], comprising a first polypeptide comprising an scFv that specifically binds to human HLA/NY-ESO, a heavy chain variable region and a CH1 constant region of an antibody that specifically binds to CD3, and an Fc region of an immunoglobulin (i) in this order from the N terminus to the C terminus, a second polypeptide that comprises a hinge region and an Fc region (ii) of an immunoglobulin, and a third polypeptide that comprises a light chain of an antibody consisting of a variable region and a constant region, wherein the first polypeptide is preferably linked to a second polypeptide in the Fc region (i) and the Fc region (ii), and the first polypeptide is preferably linked to a third polypeptide in the heavy chain variable region and the CH1 constant region of the antibody.

[56] The molecule according to [55], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [1], which is an scFv.

[57] The molecule according to [55], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [2], which is an scFv.

[58] The molecule according to [55], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [3], which is an scFv.

[59] The molecule according to [55], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [4], which is an scFv.

[60] The molecule according to [55], wherein the scFv that specifically binds to human HLA/NY-ESO is an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof according to [5], which is an scFv.

[61] The molecule according to any of [18]-[21], wherein the antibody that specifically binds to CD3 or a binding fragment thereof is a Fab.

[62] The molecule according to any one of [56]-[61], wherein the Fab that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [18], which is a Fab.

[63] The molecule according to any one of [56]-[61], wherein the Fab that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [19], which is a Fab.

[64] The molecule according to any of [56]-[61], wherein the Fab that specifically binds to CD3 is an antibody that specifically binds to CD3, or a binding fragment thereof according to [20] or [21], which is a Fab.

[65] The molecule of any one of [55]-[64], wherein the first polypeptide comprises an amino acid sequence of amino acids 21-394 of the amino acid sequence of SEQ ID NO:160.

[66] The molecule of any one of [55]-[65], wherein the first polypeptide comprises an amino acid sequence of any one of (i)-(iii) below:

(i) the amino acid sequence of amino acids 20-724 of the amino acid sequence SEQ ID NO:160;

(ii) the amino acid sequence of amino acids 20-719 of the amino acid sequence of SEQ ID NO:197; and

(iii) the amino acid sequence of amino acids 20-719 of the amino acid sequence SEQ ID NO:198.

[67] The molecule of any one of [55]-[66], wherein the second polypeptide comprises an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84.

[68] The molecule of any one of [56]-[67], wherein the third polypeptide comprises an amino acid sequence of amino acids 21-127 of the amino acid sequence of SEQ ID NO:161.

[69] The molecule of any one of [56]-[68], wherein the third polypeptide comprises an amino acid sequence of amino acids 21-233 of the amino acid sequence of SEQ ID NO:161.

[70] A molecule according to any one of paragraphs [15] to [69], wherein 1 or 2 amino acids are removed from the carboxyl terminus of the amino acid sequence of at least one polypeptide included in the molecule.

[71] A polynucleotide comprising a nucleotide sequence encoding an amino acid sequence included in the molecule of any one of [15]-[70].

[72] A vector containing the polynucleotide according to [71].

[73] A host cell containing a polynucleotide according to [71] or a vector according to [72].

[74] A method for producing a molecule that specifically binds to human HLA/NY-ESO and human CD3, comprising: (i) the step of culturing a host cell according to [73]; and (ii) the step of purifying an antibody or a binding fragment thereof from the culture product obtained in step (i).

[75] A molecule that specifically binds to human HLA/NY-ESO and human CD3, which is obtained by the method of [74].

[76] A pharmaceutical composition comprising as an active ingredient a molecule according to any one of [15]-[70] and [75], a polynucleotide according to [71], a vector according to [72], or a host cell according to [73].

[77] The pharmaceutical composition according to [14] or [76], which is an anticancer agent.

[78] The pharmaceutical composition according to [77], wherein the cancer is one or more cancers selected from the group consisting of kidney cancer, melanoma, squamous cell cancer, basal cell cancer, conjunctival cancer, oral cavity cancer, laryngeal cancer, pharyngeal cancer, thyroid cancer, lung cancer (non-small cell lung cancer (adenocarcinoma, epidermoid cancer, large cell cancer) and small cell lung cancer), breast cancer, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, colon cancer, rectal cancer, appendiceal cancer, anal cancer, liver cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, adrenal cancer, bladder cancer, prostate cancer, uterine cancer, vaginal cancer, liposarcoma, angiosarcoma, chondrosarcoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, undifferentiated pleomorphic sarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, retroperitoneal sarcoma, synovial sarcoma, uterine sarcoma, gastrointestinal stromal tumor, leiomyosarcoma, epithelioid sarcoma, B-cell lymphoma, T/NK-cell lymphoma, Hodgkin's lymphoma, myelogenous leukemia, lymphocytic leukemia, myeloproliferative disorder, myelodysplastic syndrome, multiple myeloma, testicular carcinoma, and ovarian cancer.

[79] A pharmaceutical composition according to any of [76]-[78], which is used in combination with another agent.

The present description covers the contents described in Japanese Patent Application No. 2020-061476, the priority of which is claimed by the present application.

Beneficial effects of the invention

According to the present invention, an antibody that binds to HLA-A2/NY-ESO and a novel bispecific antibody (bispecific molecule) that binds to HLA-A2/NY-ESO and to CD3 are obtained. A novel pharmaceutical composition containing such an antibody (molecule) as an active ingredient is also obtained. Such an antibody or molecule has cytotoxicity and can thus be used as an agent for the treatment or prevention of cancer and the like.

Brief description of the drawings

[Fig. 1] Fig. 1 is a table showing the standardized gMFI of anti-HLA/NY-ESO scFv: NYA-0001, NYA-1143, NYA-1154, NYA-1163, NYA-2023, NYA-2027, NYA-2035, NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060, NYA-2061, NYA-2143, NYC-0003, and NYC-0004 compared with T2 cells supplemented with different point mutant peptides. *Each underlined value indicates half or less of the standardized gMFI of each scFv compared with T2 cells supplemented with NY-ESO peptide.

[Fig. 2A] Fig. 2A is a table showing the information regarding the selected homologous peptides. The binding affinity to HLA-A0201 predicted using NetMHCPan2.8 is indicated in terms of 50% inhibitory concentration (IC ₅₀ ).

[Fig. 2B] Fig. 2B is a table showing the standardized gMFI of anti-HLA/NY-ESO scFv: NYA-0001, NYA-1143, NYA-1154, NYA-1163, NYA-2023, NYA-2027, NYA-2035, NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060, NYA-2061, NYA-2143, NYC-0003, and NYC-0004 compared to T2 cells supplemented with different homologous peptides. *Each underlined value indicates a value higher than the standardized gMFI of each scFv relative to that of T2 cells supplemented with DMSO.

[Fig. 3] Fig. 3 shows the antibody formats demonstrated in the Examples. (a) Shows the scFv format in which the variable region of the H chain of an antibody (VH, described below) is ligated to the variable region of the L chain of an antibody (VL, described below) (both empty) via a linker. In the Examples, anti-HLA-A2/NY-ESO scFv and anti-CD3 scFv were used for the evaluation. (b) Shows the Fab format, which includes VH (empty), the constant region of the H chain of an antibody (CH1: checkered pattern), VL (empty), and the constant region of the L chain of an antibody (border lines). In the Examples, anti-HLA-A2/NY-ESO Fab and the like were used for the evaluation. (c) Shows the taFv format in which two types of scFv constructs (empty and positive slopes) are ligated to each other via a linker. In the examples, taFv including anti-HLA-A2/NY-ESO scFv and anti-CD3 scFv are used for the evaluation. (d) The Fc-type format of a taFv heterodimer is shown, in which the Fc (negative slopes) containing a heterodimer-forming mutation introduced into the C-terminal site of taFv (first polypeptide) is heterologously linked to another Fc (shaded: second polypeptide). In the example, a taFv Fc heterodimer containing anti-HLA-A2/NY-ESO scFv and anti-CD3 scFv is used for the evaluation. (e) The Fc-type format of a taFv-Fab heterodimer is shown, in which a Fab is added to the Fc-type format of a taFv heterodimer. In the example, taFv, including anti-HLA-A2/NY-ESO scFv and anti-CD3 scFv, and taFv-Fab-heterodimer-Fc, including HLA-A2/NY-ESO Fab are used for the evaluation. (f) A first polypeptide common to the taFv Fc-type heterodimer and the taFv-Fab Fc-type heterodimer is shown. The first polypeptide comprises an scFv that specifically binds to human HLA/NY-ESO, an scFv that specifically binds to CD3, and an Fc region (i) in that order from N terminus to C terminus. (g) A second polypeptide of the taFv-Fc-type heterodimer is shown. The second polypeptide comprises a hinge region and an Fc region (ii). (h) A second polypeptide of the taFv-Fab-Fc-type heterodimer is shown. The second polypeptide comprises an immunoglobulin heavy chain that includes a hinge region and an Fc region (ii). (i) The third polypeptide of the taFv-Fab Fc-type heterodimer is shown. The third polypeptide contains an immunoglobulin light chain.

[Fig. 4A] Fig. 4A shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0019, NYF-0022, NYF-0023, NYF-0027, NYF-0035, NYF-0044, and NYF-0047 exhibit cytotoxicity against human U266B1 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4B] Fig. 4B shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016 and NYF-0058, exhibit cytotoxicity against human U266B1 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4C] Fig. 4C shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0023, NYF-0045, NYF-0048, NYF-0060, and NYF-0061, exerted cytotoxic effects on endogenous human NY-ESO-expressing U266B1 cells endogenously expressing NY-ESO in the presence of human PBMCs. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4D] Fig. 4D shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0019, NYF-0022, NYF-0023, NYF-0027, NYF-0035, NYF-0044, and NYF-0047 exhibit cytotoxicity against human NCI-H1703 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4E] Fig. 4E shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016 and NYF-0058, exhibit cytotoxicity against human NCI-H1703 cells endogenously expressing NY-ESO in the presence of human PBMCs. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4F] Fig. 4F shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0023, NYF-0045, NYF-0048, NYF-0060, and NYF-0061 exhibit cytotoxicity against human NCI-H1703 cells endogenously expressing NY-ESO in the presence of human PBMCs. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4G] Fig. 4G shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0019, NYF-0022, NYF-0023, NYF-0027, NYF-0035, NYF-0044, and NYF-0047, have no cytotoxic effect on human AGS cells that do not endogenously express NY-ESO in the presence of human PBMC. The error bar in the figure indicates the standard deviation (n=3).

[Fig. 4H] Fig. 4H shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016 and NYF-0058, do not exhibit cytotoxicity toward human AGS cells that do not endogenously express NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4I] Fig. 4I shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0023, NYF-0045, NYF-0048, NYF-0060, and NYF-0061, show no cytotoxicity toward human AGS cells that do not endogenously express NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4J] Fig. 4J shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0019, NYF-0022, NYF-0023, NYF-0027, NYF-0035, NYF-0044, and NYF-0047, have no cytotoxic effect on human CFPAC-1 cells that do not endogenously express NY-ESO in the presence of human PBMC. The error bar in the figure indicates the standard deviation (n=3).

[Fig. 4K] Fig. 4K shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016 and NYF-0058, do not exhibit cytotoxicity toward human CFPAC-1 cells, which do not endogenously express NY-ESO, in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 4L] Fig. 4L shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0023, NYF-0045, NYF-0048, NYF-0060, and NYF-0061, show no cytotoxicity toward human CFPAC-1 cells, which do not endogenously express NY-ESO, in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 5A] Fig. 5A shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0019, NYF-0044, and NYF-0047, exhibit antitumor activity in human PBMC-transfected models. Error bars in the figure indicate standard deviation (n=5, but only n=4 at day 32 in the NYF-0044 group).

[Fig. 5B] Fig. 5B shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0022, NYF-0023, NYF-0027, NYF-0035, and NYF-0058 exhibited antitumor activity in human PBMC-transfected models. Error bars in the figure indicate standard deviation (n=5, n=6 in vehicle control group only).

[Fig. 5C] Fig. 5C shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYF-0016, NYF-0045, NYF-0048, NYF-0060, and NYF-0061, exhibit antitumor activity in human PBMC-transfected models. Error bars in the figure indicate standard deviation (n=5).

[Fig. 6A] Fig. 6A shows the antibody formats described in the Examples. (a): Hybrid type format: Fc (oblique lines) containing a heterodimer-forming mutation and Fc (solid) containing a heterodimer-forming mutation are added to the C terminal region of Fab and scFv, respectively, and the two Fc constructs are linked. In the example, a hybrid type format containing anti-HLA-A2/NY-ESO Fab and anti-CD3 scFv is used for the evaluation. (b): Dual type format: Fc (negative slopes) containing a heterodimer-forming mutation and Fc (solid) containing a heterodimer-forming mutation are added to the C terminal regions of two different types of scFv constructs, and the two Fc constructs are heterologously linked. In the example, a dual type format containing anti-HLA-A2/NY-ESO scFv and anti-CD3 scFv is used for the evaluation. (c): scFv-Fab-Fc-type heterodimer format: Fc (negative slopes) containing a mutation that forms a heterodimer is added to the C terminal region, where the scFv is ligated with a Fab linker and linked to another Fc (solid). In the example, the scFv-Fab-Fc-type heterodimer format containing anti-CD3 scFv (positive slopes) and anti-HLA-A2/NY-ESO Fab is used for the evaluation. In addition, the scFv-Fab-Fc-type heterodimer containing anti-HLA-A2/NY-ESO scFv (positive slopes) and anti-CD3 Fab is used for the evaluation.

[Fig. 6B] Fig. 6B shows the antibody formats described in the Examples. (a): taFv-Fc-type heterodimer format: the same as in Fig. 3(d); Fc (negative slopes) containing a heterodimer-forming mutation is added to the C terminal region of taFv and heterologously linked to another Fc (solid). In the example, an Fc-type heterodimer taFv comprising an anti-HLA-A2/NY-ESO scFv and an anti-CD3 scFv is used for the evaluation. (b): taFv (inverted)-Fc-type heterodimer format: the order of the two scFv constructs is inverted in the taFv-Fc-type heterodimer format. In the example, an taFv (inverted)-Fc-type heterodimer comprising an anti-CD3 scFv and an anti-HLA-A2/NY-ESO scFv is used for the evaluation. (c) A first taFv (inverted)-Fc type heterodimer polypeptide is shown. The first polypeptide comprises an scFv that specifically binds to CD3, an scFv that specifically binds to human HLA/NY-ESO, and an Fc region (i) in that order from N terminus to C terminus. (d) A second taFv (inverted)-Fc type heterodimer polypeptide is shown. The second polypeptide comprises a hinge region and an Fc region (ii).

[Fig. 7A] Fig. 7A shows that various anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; hybrid type (NYG-3143), dual type (NYG-2143) and taFv-Fc type heterodimer (NYF-0011) exhibit cytotoxicity against human U266B1 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 7B] Fig. 7B shows that various anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; scFv-Fab-Fc type heterodimer (NYF-0003), taFv-Fc type heterodimer (NYF-0010), taFv (inverted)-Fc type heterodimer (NYF-0004), exhibited cytotoxicity against human U266B1 cells endogenously expressing NY-ESO in the presence of human PBMC. The error bar in the figure indicates the standard deviation (n=3).

[Fig. 8] Amino acid sequence of the peptide in NY-ESO (SEQ ID NO:1).

[Fig. 9] Amino acid sequence of the peptide in MAGEC-1 (SEQ ID NO:2).

[Fig. 10] Primer 1 for scFv sequence analysis (SEQ ID NO:3).

[Fig. 11] Primer 2 for scFv sequence analysis (SEQ ID NO:4).

[Fig. 12] Nucleotide sequence of the variable region of the heavy chain of NYA-0001 (SEQ ID NO:5).

[Fig. 13] Amino acid sequence of the variable region of the heavy chain of NYA-0001 (SEQ ID NO:6).

[Fig. 14] Nucleotide sequence of the light chain variable region of NYA-0001 (SEQ ID NO:7).

[Fig. 15] Amino acid sequence of the light chain variable region of NYA-0001 (SEQ ID NO:8).

[Fig. 16] Nucleotide sequence of the variable region of the heavy chain of NYA-0060 (SEQ ID NO:9).

[Fig. 17] Amino acid sequence of the variable region of the heavy chain of NYA-0060 (SEQ ID NO:10).

[Fig. 18] Nucleotide sequence of the light chain variable region of NYA-0060 (SEQ ID NO:11).

[Fig. 19] Amino acid sequence of the light chain variable region of NYA-0060 (SEQ ID NO:12).

[Fig. 20] Nucleotide sequence of the variable region of the heavy chain of NYA-0068 (SEQ ID NO:13).

[Fig. 21] Amino acid sequence of the variable region of the heavy chain of NYA-0068 (SEQ ID NO:14).

[Fig. 22] Nucleotide sequence of the light chain variable region of NYA-0068 (SEQ ID NO:15).

[Fig. 23] Amino acid sequence of the light chain variable region of NYA-0068 (SEQ ID NO:16).

[Fig. 24] Nucleotide sequence of the variable region of the heavy chain of NYA-0082 (SEQ ID NO:17).

[Fig. 25] Amino acid sequence of the variable region of the heavy chain of NYA-0082 (SEQ ID NO:18).

[Fig. 26] Nucleotide sequence of the light chain variable region of NYA-0082 (SEQ ID NO:19).

[Fig. 27] Amino acid sequence of the light chain variable region of NYA-0082 (SEQ ID NO:20).

[Fig. 28] Nucleotide sequence of the NYA-1163 tag adduct (SEQ ID NO:21).

[Fig. 29] Nucleotide sequence of the NYA-2023 tag adduct (SEQ ID NO:22).

[Fig. 30] Nucleotide sequence of the NYA-2027 tag adduct (SEQ ID NO:23).

[Fig. 31] Nucleotide sequence of the NYA-1143 tag adduct (SEQ ID NO:24).

[Fig. 32] Nucleotide sequence of the NYA-2143 tag adduct (SEQ ID NO:25).

[Fig. 33] Amino acid sequence of NYA-1163 tag adduct (SEQ ID NO:26); NYA-1163: amino acids 21-266.

[Fig. 34] Amino acid sequence of the NYA-2023 tag adduct (SEQ ID NO:27); NYA-2023: amino acids 21-266.

[Fig. 35] Amino acid sequence of the NYA-2027 tag adduct (SEQ ID NO:28); NYA-2027: amino acids 21-266.

[Fig. 36] Amino acid sequence of NYA-1143 tag adduct (SEQ ID NO:29); NYA-1143: amino acids 21-266.

[Fig. 37] Amino acid sequence of the NYA-2143 tag adduct (SEQ ID NO:30); NYA-2143: amino acids 21-266.

[Fig. 38] Nucleotide sequence of the NYA-1154 tag adduct (SEQ ID NO:31).

[Fig. 39] Amino acid sequence of the NYA-1154 tag adduct (SEQ ID NO:32); NYA-1154: amino acids 21-266.

[Fig. 40] Truncated amino acid sequence of HLA-A*0201 (GenBank: ASA47534.1) (SEQ ID NO:33).

[Fig. 41] Amino acid sequence of β2-microglobin (SEQ ID NO:34).

[Fig. 42] Nucleotide sequence of the NYA-2035 tag adduct (SEQ ID NO:35).

[Fig. 43] Amino acid sequence of the NYA-2035 tag adduct (SEQ ID NO:36); NYA-2035: amino acids 21-266.

[Fig. 44] Amino acid sequence of NYA-1143-VH01 (SEQ ID NO:37).

[Fig. 45] Amino acid sequence of NYA-1143-VH02 (SEQ ID NO:38).

[Fig. 46] Amino acid sequence of NYA-1143-VH03 (SEQ ID NO:39).

[Fig. 47] Amino acid sequence of NYA-1143-VL01 (SEQ ID NO:40).

[Fig. 48] Nucleotide sequence of the NYA-2044 tag adduct (SEQ ID NO:41).

[Fig. 49] Nucleotide sequence of the NYA-2045 tag adduct (SEQ ID NO:42).

[Fig. 50] Nucleotide sequence of the NYA-2047 tag adduct (SEQ ID NO:43).

[Fig. 51] Nucleotide sequence of the NYA-2048 tag adduct (SEQ ID NO:44).

[Fig. 52] Nucleotide sequence of the NYA-2060 tag adduct (SEQ ID NO:45).

[Fig. 53] Nucleotide sequence of the NYA-2061 tag adduct (SEQ ID NO:46).

[Fig. 54] Amino acid sequence of the NYA-2044 tag adduct (SEQ ID NO:47); NYA-2044: amino acids 21-266.

[Fig. 55] Amino acid sequence of the NYA-2045 tag adduct (SEQ ID NO:48); NYA-2045: amino acids 21-266.

[Fig. 56] Amino acid sequence of NYA-0082 (SEQ ID NO:49).

[Fig. 57] Amino acid sequence of the NYA-2047 tag adduct (SEQ ID NO:50); NYA-2047: amino acids 21-266.

[Fig. 58] Amino acid sequence of the NYA-2048 tag adduct (SEQ ID NO:51); NYA-2048: amino acids 21-266.

[Fig. 59] Amino acid sequence of the NYA-2060 tag adduct (SEQ ID NO:52); NYA-2060: amino acids 21-266.

[Fig. 60] Amino acid sequence of the NYA-2061 tag adduct (SEQ ID NO:53); NYA-2061: amino acids 21-266.

[Fig. 61] Amino acid sequences of CDRH1 to CDRH3 of the heavy chain of NYA-0001 and CDRL1 to CDRL3 of the light chain of NYA-0001 (SEQ ID NOs:54-59).

[Fig. 62] Amino acid sequence of CDRL1 NYA-2023 (SEQ ID NO:60).

[Fig. 63] Amino acid sequence of CDRL3 NYA-2027 (SEQ ID NO:61).

[Fig. 64] Amino acid sequences of CDRH3 and CDRL3 of NYA-1154 (SEQ ID NOs:62 and 63).

[Fig. 65] Amino acid sequence of CDRL1 NYA-0035 (SEQ ID NO:64).

[Fig. 66] Nucleotide sequence of the NYC-0003 tag adduct (SEQ ID NO:65).

[Fig. 67] Nucleotide sequence of the NYC-0004 tag adduct (SEQ ID NO:66).

[Fig. 68] Amino acid sequence of the NYC-0003 tag adduct (SEQ ID NO:67); NYC-0003: amino acids 21 to 263.

[Fig. 69] Amino acid sequence of the NYC-0004 tag adduct (SEQ ID NO:68); NYC-0004: amino acids 21 to 263.

[Fig. 70] Nucleotide sequence of the NYA-0001 tag adduct (SEQ ID NO:69).

[Fig. 71] Amino acid sequence of the NYA-0001 tag adduct (SEQ ID NO:70); NYA-0001: amino acids 21-266.

[Fig. 72] Nucleotide sequence of HC1 (SEQ ID NO:71).

[Fig. 73] Nucleotide sequence of NYF-0016-HC2 (SEQ ID NO:72).

[Fig. 74] Nucleotide sequence of NYF-0019-HC2 (SEQ ID NO:73).

[Fig. 75] Nucleotide sequence of NYF-0022-HC2 (SEQ ID NO:74).

[Fig. 76] Nucleotide sequence of NYF-0023-HC2 (SEQ ID NO:75).

[Fig. 77] Nucleotide sequence of NYF-0027-HC2 (SEQ ID NO:76).

[Fig. 78] Nucleotide sequence of NYF-0035-HC2 (SEQ ID NO:77).

[Fig. 79] Nucleotide sequence of NYF-0044-HC2 (SEQ ID NO:78).

[Fig. 80] Nucleotide sequence of NYF-0045-HC2 (SEQ ID NO:79).

[Fig. 81] Nucleotide sequence of NYF-0047-HC2 (SEQ ID NO:80).

[Fig. 82] Nucleotide sequence of NYF-0048-HC2 (SEQ ID NO:81).

[Fig. 83] Nucleotide sequence of NYF-0060-HC2 (SEQ ID NO:82).

[Fig. 84] Nucleotide sequence of NYF-0061-HC2 (SEQ ID NO:83).

[Fig. 85] Amino acid sequence of HC1 (SEQ ID NO:84).

[Fig. 86] Amino acid sequence of NYF-0016-HC2 (SEQ ID NO:85); NYA-1143: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 87] Amino acid sequence of NYF-0019-HC2 (SEQ ID NO:86); NYA-2143: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 88] Amino acid sequence of NYF-0022-HC2 (SEQ ID NO:87); NYA-1163: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 89] Amino acid sequence of NYF-0023-HC2 (SEQ ID NO:88); NYA-2023: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 90] Amino acid sequence of NYF-0027-HC2 (SEQ ID NO:89); NYA-2027: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 91] Amino acid sequence of NYF-0035-HC2 (SEQ ID NO:90); NYA-2035: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 92] Amino acid sequence of NYF-0044-HC2 (SEQ ID NO:91); NYA-2044: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 93] Amino acid sequence of NYF-0045-HC2 (SEQ ID NO:92); NYA-2045: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 94] Amino acid sequence of NYF-0047-HC2 (SEQ ID NO:93); NYA-2047: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 95] Amino acid sequence of NYF-0048-HC2 (SEQ ID NO:94); NYA-2048: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 96] Amino acid sequence of NYF-0060-HC2 (SEQ ID NO:95); NYA-2060: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 97] Amino acid sequence of NYF-0061-HC2 (SEQ ID NO:96); NYA-2061: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 98] Nucleotide sequence of the NYA-0001-Fab-HC1-k deletion (SEQ ID NO:97)

[Fig. 99] Nucleotide sequence of NYA-0001-LC (SEQ ID NO:98).

[Fig. 100] Amino acid sequence of the NYA-0001-Fab-HC1-k deletion (SEQ ID NO:99); NYA-0001 heavy chain variable region: amino acids 20-139.

[Fig. 101] Amino acid sequence of NYA-0001-LC (SEQ ID NO:100); NYA-0001 light chain variable region: amino acids 21-131.

[Fig. 102] Nucleotide sequence of the NYA-1143-Fab-HC1-k deletion (SEQ ID NO:101).

[Fig. 103] Nucleotide sequence of NYA-1143-LC (SEQ ID NO:102).

[Fig. 104] Nucleotide sequence of C3E-7085-HC2-k with C deletion (SEQ ID NO:103).

[Fig. 105] Amino acid sequence of NYA-1143-Fab-HC1-k deletion (SEQ ID NO:104); NYA-1143 heavy chain variable region: amino acids 20-139.

[Fig. 106] Amino acid sequence of NYA-1143-LC (SEQ ID NO:105); NYA-1143 light chain variable region: amino acids 21-131.

[Fig. 107] Amino acid sequence of the C3E-7085-HC2-k deletion (SEQ ID NO:106); C3E-7085: amino acids 21-260.

[Fig. 108] Nucleotide sequence of the NYA-1143-HC1-k deletion (SEQ ID NO:107).

[Fig. 109] Amino acid sequence of the NYA-1143-HC1-k deletion (SEQ ID NO:108); NYA-1143: amino acids 21-266.

[Fig. 110] Nucleotide sequence of the C3E-7085-NYA-1154-Fab-HC2-k deletion (SEQ ID NO:109).

[Fig. 111] Nucleotide sequence of NYA-1154-LC (SEQ ID NO:110).

[Fig. 112] Nucleotide sequence of the OAA-HC1-k deletion (SEQ ID NO:111).

[Fig. 113] Amino acid sequence of the deletion C3E-7085-NYA-1154-Fab-HC2-k (SEQ ID NO:112); C3E-7085: amino acids 21-260; heavy chain variable region NYA-1154: amino acids 266-285.

[Fig. 114] Amino acid sequence of NYA-1154-LC (SEQ ID NO:113); NYA-1154 light chain variable region: amino acids 21-131.

[Fig. 115] Amino acid sequence of the OAA-HC1-k deletion (SEQ ID NO:114).

[Fig. 116] Nucleotide sequence of the NYF-0010-HC2-k deletion (SEQ ID NO:115).

[Fig. 117] Nucleotide sequence of the NYF-0004-HC2-k deletion (SEQ ID NO:116).

[Fig. 118] Nucleotide sequence of the NYF-0011-HC2-k deletion (SEQ ID NO:117).

[Fig. 119] Amino acid sequence of the NYF-0010-HC2-k deletion (SEQ ID NO:18); NYA-1154: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 120] Amino acid sequence of the NYF-0004-HC2-k deletion (SEQ ID NO:119); C3E-7085: amino acids 21-260; NYA-1154: amino acids 272-511.

[Fig. 121] Amino acid sequence of the NYF-0011-HC2-k deletion (SEQ ID NO:120); NYA-1143: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 122] Amino acid sequence of the point mutant peptide 1F NY-ESO (SEQ ID NO:121).

[Fig. 123] Amino acid sequence of the point mutant peptide NY-ESO 2M (SEQ ID NO:122).

[Fig. 124] Amino acid sequence of the point mutant peptide 3A NY-ESO (SEQ ID NO:123).

[Fig. 125] Amino acid sequence of the point mutant peptide 4A NY-ESO (SEQ ID NO:124).

[Fig. 126] Amino acid sequence of the point mutant peptide 5A NY-ESO (SEQ ID NO:125).

[Fig. 127] Amino acid sequence of the point mutant peptide 6L NY-ESO (SEQ ID NO:126).

[Fig. 128] Amino acid sequence of the point mutant peptide 7F NY-ESO (SEQ ID NO:127).

[Fig. 129] Amino acid sequence of the point mutant peptide 8A NY-ESO (SEQ ID NO:128).

[Fig. 130] Amino acid sequence of the point mutant peptide 9A NY-ESO (SEQ ID NO:129).

[Fig. 131] Amino acid sequence of the gp100 peptide (SEQ ID NO:130).

[Fig. 132] Amino acid sequence of the DOLPP1 homologous peptide (SEQ ID NO:131).

[Fig. 133] Amino acid sequence of the IL20RB homologous peptide (SEQ ID NO:132).

[Fig. 134] Amino acid sequence of PRKD2 homologous peptide (SEQ ID NO:133).

[Fig. 135] Amino acid sequence of the homologous peptide of CD163 (SEQ ID NO:134).

[Fig. 136] Amino acid sequence of the homologous peptide P2RY8 (SEQ ID NO:135).

[Fig. 137] Amino acid sequence of C3E-7034 (SEQ ID NO:136).

[Fig. 138] Amino acid sequence of C3E-7036 (SEQ ID NO:137).

[Fig. 139] Amino acid sequence of C3E-7085 (SEQ ID NO:138).

[Fig. 140] Amino acid sequence of C3E-7088 (SEQ ID NO:139).

[Fig. 141] Amino acid sequence of C3E-7093 (SEQ ID NO:140).

[Fig. 142] Amino acid sequences of CDRH1-CDRH3 of the heavy chain of C3E-7085 and CDRL1-CDRL3 of the light chain of C3E-7085 (SEQ ID NOs:141-146).

[Fig. 143] Amino acid sequence of C3E-7078 (SEQ ID NO:147).

[Fig. 144] Nucleotide sequence of NYF-0014-HC2 (SEQ ID NO:148).

[Fig. 145] Amino acid sequence of NYF-0014-HC2 (SEQ ID NO:149); NYA-0001: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 146] Amino acid sequence of NYF-0082-HC2 (SEQ ID NO:150); NYA-0082: amino acids 21-266; C3E-7085: amino acids 272-511.

[Fig. 147] Amino acid sequence of human CD3 ε (SEQ ID NO:151).

[Fig. 148] Nucleotide sequence of entire NYZ-0038-HC2 (SEQ ID NO:152).

[Fig. 149] Nucleotide sequence of entire NYZ-0082-HC2 (SEQ ID NO:153).

[Fig. 150] Nucleotide sequence of entire NYZ-0083-HC2 (SEQ ID NO:154).

[Fig. 151] Amino acid sequence of the entire NYZ-0038-HC2 (SEQ ID NO:155); NYA-2061: amino acids 21-266; C3E-7096: amino acids 272-511.

[Fig. 152] Amino acid sequence of the entire NYZ-0082-HC2 (SEQ ID NO:156); NYA-3061: amino acids 21-271; C3E-7096: amino acids 277-516.

[Fig. 153] Amino acid sequence of the entire NYZ-0083-HC2 (SEQ ID NO:157); NYA-3061: amino acids 21-271; C3E-7097: amino acids 277-516.

[Fig. 154] Nucleotide sequence of full-length NYZ-1010-HC2 (SEQ ID NO:158).

[Fig. 155] Nucleotide sequence of full-length C3E-7085-LC (SEQ ID NO:159).

[Fig. 156] Amino acid sequence of full-length NYZ-1010-HC2 (SEQ ID NO:160); NYA-3061: amino acids 21-271; heavy chain variable region C3E-7085: amino acids 277-394.

[Fig. 157] Amino acid sequence of full-length C3E-7085-LC (SEQ ID NO:161).

[Fig. 158A] Fig. 158A is a table showing the standardized gMFI of the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; NYZ-0038, NYZ-0082, NYZ-0083, and NYZ-1010 relative to CD3e knockout T2 cells complemented with different point mutant peptides. *Each underlined value indicates one-half or less of the standardized gMFI of each Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule relative to CD3e knockout T2 cells complemented with the NY-ESO peptide, and *each bold value indicates one-quarter or less of the standardized gMFI.

[Fig. 158B] Fig. 158B is a table showing the standardized Fc gMFI of conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; NYZ-0038, NYZ-0082, NYZ-0083, and NYZ-1010 relative to CD3e knockout T2 cells supplemented with various homologous peptides. *Each underlined value indicates the value of each Fc-supplemented anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule that is higher than the standardized gMFI of CD3e knockout T2 cells supplemented with DMSO.

[Fig. 159A] Fig. 159A shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0038, NYZ-0082, and NYZ-0083, exhibit cytotoxicity against human U266B1 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 159B] Fig. 159B shows that the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule; i.e., NYZ-1010, exhibits cytotoxicity against human U266B1 cells endogenously expressing NY-ESO in the presence of human PBMC. The error bar in the figure indicates the standard deviation (n=3).

[Fig. 159C] Fig. 159C shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0082 and NYZ-1010, exhibit cytotoxicity against human NCI-H1703 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 159D] Fig. 159D shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0038 and NYZ-0083, exhibit cytotoxicity against human NCI-H1703 cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 159E] Fig. 159E shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0038, NYZ-0082, and NYZ-0083, do not exhibit cytotoxicity toward human AGS cells endogenously expressing NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 159F] Fig. 159F shows that the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule; i.e., NYZ-1010, does not exert cytotoxic effect on human AGS cells that do not endogenously express NY-ESO in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 159G] Fig. 159G shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0082 and NYZ-1010, do not exhibit cytotoxicity against human CFPAC-1 cells, which do not endogenously express NY-ESO, in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 159H] Fig. 159H shows that Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0038 and NYZ-0083, do not exhibit cytotoxicity against human CFPAC-1 cells, which do not endogenously express NY-ESO, in the presence of human PBMC. Error bars in the figure indicate standard deviation (n=3).

[Fig. 160A] Fig. 160A shows that the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule; i.e., NYZ-0038, exhibits antitumor activity in human PBMC-transfected models. The error bar in the figure indicates the standard deviation (n=5).

[Fig. 160B] Fig. 160B shows that the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule; i.e., NYZ-0082, exhibits antitumor activity in human PBMC-transfected models. The error bar in the figure indicates the standard deviation (n=5).

[Fig. 160C] Fig. 160C shows that the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule; i.e., NYZ-0083, exhibits antitumor activity in human PBMC-transfected models. The error bar in the figure indicates the standard deviation (n=5).

[Fig. 160D] Fig. 160D shows that the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule; i.e., NYZ-1010, exhibits antitumor activity in human PBMC-transfected models. The error bar in the figure indicates the standard deviation (n=5).

[Fig. 161] Amino acid sequence of the peptide linker (SEQ ID NO:162).

[Fig. 162] Fig. 162 shows the polymer content (%) upon acid treatment of various Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecules.

[Fig. 163] Fig. 163 shows the results of the stability evaluation of the anti-HLA/NY-ESO scFv-Fc heterodimer solution.

[Fig. 164] Nucleotide sequence of full-length NYA-3061 (SEQ ID NO:163).

[Fig. 165] Amino acid sequence of full-length NYA-3061 (SEQ ID NO:164).

[Fig. 166] Nucleotide sequence of full-length NYC-0005 (SEQ ID NO:165).

[Fig. 167] Amino acid sequence of full-length NYC-0005 (SEQ ID NO:166).

[Fig. 168] Nucleotide sequence of full-length NYC-0006 (SEQ ID NO:167).

[Fig. 169] Amino acid sequence of full-length NYC-0006 (SEQ ID NO:168).

[Fig. 170] Nucleotide sequence of full-length NYC-0007 (SEQ ID NO:169).

[Fig. 171] Amino acid sequence of full-length NYC-0007 (SEQ ID NO:170).

[Fig. 172] Nucleotide sequence of full-length NYC-0008 (SEQ ID NO:171).

[Fig. 173] Amino acid sequence of full-length NYC-0008 (SEQ ID NO:172).

[Fig. 174] Nucleotide sequence of full-length NYC-0009 (SEQ ID NO:173).

[Fig. 175] Amino acid sequence of full-length NYC-0009 (SEQ ID NO:174).

[Fig. 176] Nucleotide sequence of full-length NYC-0010 (SEQ ID NO:175).

[Fig. 177] Amino acid sequence of full-length NYC-0010 (SEQ ID NO:176).

[Fig. 178] Nucleotide sequence of full-length HC-h (SEQ ID NO:177).

[Fig. 179] Amino acid sequence of full-length HC-h (SEQ ID NO:178).

[Fig. 180] Nucleotide sequence of full-length NYD-2047-HC-k (SEQ ID NO:179).

[Fig. 181] Amino acid sequence of full-length NYD-2047-HC-k (SEQ ID NO:180).

[Fig. 182] Nucleotide sequence of full-length NYD-2061-HC-k (SEQ ID NO:181).

[Fig. 183] Amino acid sequence of full-length NYD-2061-HC-k (SEQ ID NO:182).

[Fig. 184] Nucleotide sequence of full-length NYD-3061-HC-k (SEQ ID NO:183).

[Fig. 185] Amino acid sequence of full-length NYD-3061-HC-k (SEQ ID NO:184).

[Fig. 186] Nucleotide sequence of full-length NYC-0011-HC-k (SEQ ID NO:185).

[Fig. 187] Amino acid sequence of full-length NYC-0011-HC-k (SEQ ID NO:186).

[Fig. 188] Nucleotide sequence of full-length NYC-0012-HC-k (SEQ ID NO:187).

[Fig. 189] Amino acid sequence of full-length NYC-0012-HC-k (SEQ ID NO:188).

[Fig. 190] Nucleotide sequence of full-length NYC-0013-HC-k (SEQ ID NO:189).

[Fig. 191] Amino acid sequence of full-length NYC-0013-HC-k (SEQ ID NO:190).

[Fig. 192] Nucleotide sequence of full-length NYC-0014-HC-k (SEQ ID NO:191).

[Fig. 193] Amino acid sequence of full-length NYC-0014-HC-k (SEQ ID NO:192).

[Fig. 194] Nucleotide sequence of full-length NYC-0015-HC-k (SEQ ID NO:193).

[Fig. 195] Amino acid sequence of full-length NYC-0015-HC-k (SEQ ID NO:194).

[Fig. 196] Nucleotide sequence of full-length NYC-0016-HC-k (SEQ ID NO:195).

[Fig. 197] Amino acid sequence of full-length NYC-0016-HC-k (SEQ ID NO:196).

[Fig. 198] Amino acid sequence of full-length NYZ-1007-HC2 (SEQ ID NO:197); NYA-2061: amino acids 21-266; heavy chain variable region C3E-7085: amino acids 272-389.

[Fig. 199] Amino acid sequence of full-length NYZ-1017-HC2 (SEQ ID NO:198); NYA-2047: amino acids 21-266; heavy chain variable region C3E-7085: amino acids 277-389.

Description of embodiments

The present invention is now described in detail.

1. Definition

In the present invention, the term "gene" refers to a nucleotide chain comprising a nucleotide sequence encoding an amino acid of a protein or its complementary chain. For example, a polynucleotide, oligonucleotide, DNA, mRNA, cDNA, cRNA or the like, which is a nucleotide chain comprising a nucleotide sequence encoding an amino acid of a protein or its complementary chain, are included within the scope of "gene". Such a gene is a single-stranded, double-stranded, triple-stranded or more stranded nucleotide, and a combination of a DNA strand and an RNA strand, a single-stranded nucleotide chain comprising both a ribonucleotide (RNA) and a deoxyribonucleotide (DNA), and a double-stranded or triple-stranded nucleotide comprising such a nucleotide chain are included within the scope of "gene". In the present invention, the term "base sequence" is synonymous with the term "nucleotide sequence".

In the present invention, the terms "polynucleotide", "nucleotide chain", "nucleic acid" and "nucleic acid molecule" are synonymous. For example, DNA, RNA, probe, oligonucleotide and primer are included in the concept of "polynucleotide". Such a polynucleotide consists of 1, 2, 3 or more chains, as well as a combination of a DNA chain and an RNA chain, a single polynucleotide chain including both a ribonucleotide (RNA) and a deoxyribonucleotide (DNA), and a double-stranded or triple-stranded nucleotide containing such a polynucleotide chain are included in the scope of "polynucleotide".

In the present invention, the terms "polypeptide", "peptide" and "protein" are synonymous.

In the present invention, the term “antigen” may refer to the term “immunogen” as appropriate.

In the present invention, the term "cell" includes various cells derived from animals, subculture cells, primary culture cells, cell lines, recombinant cells and microorganisms.

In the present invention, the term "antibody" is synonymous with the term "immunoglobulin". However, the term "antibody" in the case of the anti-HLA/NY-ESO antibody of the present invention refers to an immunoglobulin comprising a constant region and a variable region. The antibody is not particularly limited, and it may be a naturally occurring or partially or completely synthesized immunoglobulin. The anti-HLA/NY-ESO antibody of the present invention is included in the term "molecule" described below.

In the present invention, “NY-ESO peptide” means a peptide consisting of 9 amino acids 157-165 of NY-ESO-1 and LAGE-1 (SLLMWITQC: SEQ ID NO:1).

In the present invention, “HLA-A2/NY-ESO” refers to a complex of NY-ESO peptide and human leukocyte histocompatibility antigen-A2 (HLA-A2), and it is also referred to as “HLA/NY-ESO”.

In the present invention, the term "anti-HLA-A2/NY-ESO antibody" refers to an antibody that binds to HLA-A2/NY-ESO. In other words, this term refers to an antibody that recognizes HLA-A2/NY-ESO. In addition, the term "anti-HLA-A2/NY-ESO scFv" refers to an scFv that binds to HLA/NY-ESO. In other words, this term refers to an scFv that recognizes HLA-A2/NY-ESO. The terms "anti-HLA-A2/NY-ESO antibody" and "anti-HLA-A2/NY-ESO scFv" are also referred to as "anti-HLA/NY-ESO antibody" and "anti-HLA/NY-ESO scFv", respectively.

The basic four-chain structure of an antibody consists of two identical light chains (L chains) and two identical heavy chains (H chains). A light chain is linked to a heavy chain by a single covalent disulfide bond. The two heavy chains are linked to each other by one or more disulfide bonds according to the heavy chain isotypes. Each of the light chain and heavy chain has an intrachain disulfide bond at regular intervals. The light chain and heavy chain have a constant region showing very high amino acid sequence similarity and a variable region showing low amino acid sequence similarity. The light chain has a variable region (VL) at its amino terminus adjacent to a constant region (CL). The heavy chain has a variable region (VL) at its amino terminus adjacent to 3 constant regions (CH1/CH2/CH3). VL pairs with VH, and CL aligns with the first constant region of the heavy chain (CH1). VL pairs with VH to form a single antigen-binding site.

The constant region of the antibody of the present invention is not particularly limited. The antibody of the present invention for treating or preventing human diseases preferably comprises a constant region of a human antibody. Examples of constant regions of a heavy chain of a human antibody include Cγ1, Cγ2, Cγ3, Cγ4, Cµ, Cδ, Cα1, Cα2 and Cε. Examples of constant regions of a light chain of a human antibody include Cκ and Cλ.

Fab contains the heavy chain VH, adjacent CH1, light chain VL and adjacent CL. Each of the VH and VL contains a complementarity determining region (CDR).

Fc (also referred to as "Fc region") is the carboxy terminal region of the constant region of the heavy chain, it contains CH2 and CH3, and is a dimer. Fc of the present invention may comprise a naturally occurring sequence, or it may comprise a sequence obtained from a naturally occurring sequence by mutation (referred to as "mutant Fc"). In the multispecific molecule and bispecific molecule of the present invention, the Fc region is preferably a mutant Fc, and more preferably a combination of Fc regions capable of forming a heterodimer. An example of a combination of Fc regions is a combination of Fc (i) in a first polypeptide and Fc (ii) in a second polypeptide described below. The combination is not limited thereto as long as such a combination of Fc regions is capable of association (forming a heterodimer).

Examples of the mutant Fc include, but are not limited to, a modified Fc region incorporated into a heteropolymer with improved stability (including a heterodimeric Fc region) described in WO 2013/063702, an Fc comprising an immunoglobulin CD3 region induced from an IgG antibody with a knob and a trough contained in a heteropolymer described in WO 1996/27011, an Fc comprising a CH3 domain incorporated into a heterodimer that becomes electrostatically favorable upon replacement of one or more amino acids with charged amino acids described in WO 2009/089004, a heterodimeric Fc region incorporated into a heterodimer comprising a spatial mutation and/or a pI (isoelectric point) mutation described in WO 2014/110601, and a heterodimeric Fc, comprising a CH3 domain with a modification to eliminate or reduce binding to protein A, described in WO 2010/151792.

The variable region consists of a region of extreme variability called the hypervariable region (HVR) and relatively non-variable regions called framework regions (FR) separated by HVRs. The naturally occurring variable regions of the heavy and light chains contain 4 FRs connected by 3 hypervariable regions, where the hypervariable region of each chain and the hypervariable region of the other chains are very close to them, and such regions contribute to the formation of the antigen-binding site of the antibody.

It is known that the heavy chain and the light chain of the antibody molecule contain 3 complementarity determining regions (CDR). The complementarity determining region is also called a hypervariable region, it is present in the variable regions of the heavy chain and light chain of the antibody, where the variability of the primary structure is particularly high, and, as a rule, it is divided into 3 positions in the primary structure of the polypeptide chain of the heavy chain and the light chain. In the present invention, the complementarity determining regions of the heavy chain of the antibody are designated as CDRH1, CDRH2 and CDRH3 from the amino terminus of the amino acid sequence of the heavy chain, and the complementarity determining regions of the light chain are designated as CDRL1, CDRL2 and CDRL3 from the amino terminus of the amino acid sequence of the light chain. These regions are adjacent to each other in the spatial structure and determine the specificity to the antigens to which they bind.

In the present invention, the position and length of CDRs are determined according to the IMGT definition (Developmental and Comparative Immunology 27, 2003, 55-77).

FR is a variable region distinct from CDR. Usually, a variable region includes 4 FRs; i.e., FR1, FR2, FR3, and FR4.

The CDRs and FRs included in the heavy and light chains are presented in the order FRH1-CDRH1-FRH2-CDRH2-FRH3-CDRH3-FRH4 and FRL1-CDRL1-FRL2-CDRL2-FRL3-CDRL3-FRL4, respectively, from the amino terminus to the carboxyl terminus.

The CDR and FR positions can be defined according to various definitions well known in the art, such as the Kabat, Chothia, AbM, Contact definitions, in addition to IMGT.

In the present invention, the term "antigen-binding fragment of an antibody" refers to a partial fragment of an antibody having antigen-binding activity, which consists of a heavy chain variable region and a light chain variable region. Examples of the "antigen-binding fragment of an antibody" include, but are not limited to, antigen-binding fragments such as Fab, F(ab') ₂ , scFv, Fab', Fv and single-domain antibody (sdAb). Such an antigen-binding fragment of an antibody can be obtained by treating a full-length antibody protein molecule with an enzyme such as papain or pepsin, or it can be a recombinant protein produced in an adequate host cell using a recombinant gene. In the present invention, the term "antibody binding fragment" is synonymous with the term "antigen-binding fragment of an antibody".

In the present invention, the "site" to which the antibody binds; i.e. the "site" that is recognized by the antibody, is a partial peptide or a partial higher order structure of the antigen to which the antibody binds or which is recognized by the antibody.

In the present invention, such a site is called an epitope or an antibody binding site. In the present invention, a "mutant antibody" refers to a polypeptide having an amino acid sequence obtained from the amino acid sequence of a parent antibody by substitution, deletion or addition ("addition" includes "insertion") (hereinafter collectively referred to as "mutation") of amino acids and binding to HLA/NY-ESO of the present invention. The number of mutant amino acids in such a mutant antibody is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25, 30, 40 or 50. Such a mutant antibody is included in the scope of the term "antibody" of the present invention.

In the present invention, the term “multiple” in “one or more” means from 2 to 10.

As used herein, the term "molecule" means a molecule comprising an antibody or antigen-binding fragment of an antibody as described above. In addition, the term "molecule" encompasses a multispecific molecule formed from an antibody or a plurality of antigen-binding fragments derived therefrom.

The term "multispecific molecule" is not particularly limited, provided that such a molecule is capable of binding to a plurality of different epitopes on a molecule and/or to different epitopes on two or more molecules. A multispecific molecule includes an antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL). Examples of such a multispecific molecule include, but are not limited to, a full-length antibody molecule comprising two or more different types of heavy chains and light chains; i.e., a multispecific molecule of the IgG type and a molecule comprising an antigen-binding fragment consisting of two or more types of VL and VH; i.e., a molecule derived from a combination of Fab, Fab', Fv, scFv, sdAb or the like, such as a tandem scFv, diabody, single-chain diabody or triabody. In addition, a molecule obtained by genetically or chemically ligating a protein without an immunoglobulin backbone and capable of binding to an antigen with an antigen-binding fragment is included in the scope of a multispecific molecule. In the present invention, such a multispecific molecule may be called a "multispecific antibody", except for the case where the molecule does not contain an antibody or an antigen-binding fragment thereof, as required.

The anti-HLA/NY-ESO antibody of the present invention, the antigen-binding fragment of such an antibody or the molecule of the present invention have, for example, biological activity and physicochemical properties (which may also be referred to as physical properties). Specific examples thereof include various biological activities such as cytotoxicity, ADCC activity or antitumor activity (described below), and physical properties such as antigen- or epitope-binding activity, stability during production or storage and thermal stability.

In the present invention, "hybridization under stringent conditions" is carried out in a solution containing 5× SSC at 65°C, in an aqueous solution containing 2× SSC and 0.1% SDS at 65°C for 20 minutes, in an aqueous solution containing 0.5× SSC and 0.1% SDS at 65°C for 20 minutes, or in an aqueous solution containing 0.2× SSC and 0.1% SDS at 65°C for 20 minutes under washing conditions or equivalent conditions. SSC is an aqueous solution of 150 mM NaCl and 15 mM sodium citrate, and n× SSC is n-fold concentrated SSC.

In the present invention, "cell damage" is any form of pathological change that occurs in a cell. In addition to external damage, the term "cell damage" refers to any damage caused to cellular structure or function, such as DNA cleavage, nucleotide dimer formation, chromosome segregation, mitotic apparatus damage, or decreased activity of various enzymes. In the present invention, "cytotoxicity" is the induction of cell damage.

In the present invention, the term "antibody-dependent cellular cytotoxicity (ADCC)" refers to the activity of NK cells in causing antibody-mediated damage to target cells, such as tumor cells.

In the present invention, the term "T-cell redirecting cytotoxicity" refers to the induction of cytotoxicity mediated by a multispecific molecule comprising an antibody against a tumor antigen and an anti-HLA/NY-ESO antibody. Specifically, the antibody against a tumor antigen binds to a target tumor cell, the anti-HLA/NY-ESO antibody binds to a T cell, the distance between the target tumor cell and the T cell is reduced, and cytotoxicity is induced by T cell activation. Such a molecule can be included in a pharmaceutical composition.

2. Antigen

2-1. HLA/NY-ESO antigen

In the present invention, “HLA/NY-ESO” is used in the same sense as “HLA/NY-ESO protein”.

HLA/NY-ESO is a ternary complex of HLA-A2, β2-microglobulin, and NY-ESO peptide. HLA-A2 is the HLA allele that is most frequently expressed in Caucasians. HLA forms a ternary complex with β2-microglobulin and a peptide fragment of an autologous protein in the cellular endoplasmic reticulum; the complex is presented extracellularly and is recognized by the T cell receptor (TCR) of the T cell. NY-ESO peptide (SLLMWITQC: SEQ ID NO:1, Fig. 8) is reported to be a peptide consisting of 9 amino acids at positions 157-165 of NY-ESO-1 and LAGE-1, which is presented by HLA-A2.

2-2. CD3 antigen

In the present invention, “CD3” is used in the same sense as “CD3 protein”.

CD3 is expressed on the T cell as part of the multimolecular T cell receptor complex, and is a complex of 5 types of polypeptide chains γ, δ, ε, ζ and η (their molecular weights are 25,000-28,000, 21,000, 20,000, 16,000 and 22,000, respectively).

Examples of CD3 complexes include the γ, δ, ε, ζ, and η chains, which are also called subunits. When an anti-CD3 antibody binds to a T cell, cell damage is induced by T cell activation. Many anti-CD3 antibodies bind to CD3 ε.

The nucleotide sequence of the cDNA encoding human CD3 ε is registered under the accession number: NM_000733 (NM_000733.3) in NCBI/GenBank, and the amino acid sequence of human CD3 ε is registered under the accession number: NP_000724 (NM_000724.1) in NCBI/GenPept. The nucleotide sequence of the cDNA encoding cynomolgus monkey CD3 is registered under the accession number: NM_001283615.1 in GenBank. The amino acid sequence of human CD3 ε is shown in SEQ ID NO: 151 of the sequence listing (Fig. 147).

2-3. Obtaining antigen

The antigen proteins used in the present invention are described above; i.e., HLA/NY-ESO and CD3 (hereinafter HLA/NY-ESO and CD3 are collectively referred to as antigen proteins) can be obtained from animal tissues (including biological fluids), cells obtained from animal tissues, or a cell culture product by, for example, purification, isolation, gene recombination, in vitro translation, or chemical synthesis.

The cDNA of the antigenic protein can be obtained by the so-called PCR method, in which the polymerase chain reaction (hereinafter referred to as “PCR”) is carried out using, as a template, a cDNA library of organs expressing mRNA of the antigenic protein and primers that specifically amplify the cDNA of the antigenic protein (Saiki, R. K., et al., Science, 1988, 239, 487-49).

The cDNA of an antigenic protein comprises a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of a nucleotide sequence complementary to a nucleotide sequence encoding an antigenic protein expressed in a human or rat and encoding a protein that has a biological activity equivalent to the biological activity of the antigenic protein.

In addition, the cDNA of the antigenic protein includes a splice variant transcribed from the locus of the antigenic protein expressed in a human or rat and a polynucleotide that hybridizes with it under stringent conditions and codes for a protein that has a biological activity equivalent to that of the antigenic protein.

A nucleotide sequence encoding a protein comprising the amino acid sequence of a human or rat antigenic protein, or an amino acid sequence obtained from such an amino acid sequence by substitution, deletion or addition of one or more amino acids from which the signal sequence has been removed, and having a biological activity equivalent to the activity of the antigenic protein, is included in the nucleotide sequence of the antigenic protein gene.

A protein comprising an amino acid sequence encoded by a splice variant transcribed from a human or rat antigenic protein gene locus, or an amino acid sequence derived from such an amino acid sequence by substitution, deletion or addition of one or more amino acids, and having a biological activity equivalent to that of the antigenic protein, is included in the antigenic protein.

2-4. Specificity of binding to antigen protein

The anti-HLA/NY-ESO antibody of the present invention, its antigen-binding fragment and the like recognize HLA/NY-ESO. In particular, they bind to the HLA/NY-ESO antigen. The presence of HLA/NY-ESO in animals other than humans, such as mice, rats and cynomolgus monkeys, is unknown.

The anti-CD3 antibody included in the multispecific molecule of the present invention, its binding fragments and the like recognize; i.e., bind to the CD3 antigen. Such an anti-CD3 antibody and the like preferably binds, for example, to human CD3 and monkey CD3, and more preferably to human CD3 and cynomolgus monkey CD3. In contrast, such a preferred anti-CD3 antibody does not bind to rat and/or mouse CD3.

The antitumor activity of the multispecific molecule of the present invention can be evaluated, for example, by (i) transplanting human cancer cells or human cancer tissue into non-human animals to which human peripheral blood lymphocytes have been transplanted, preferably rats or mice, and more preferably rats or mice deficient in endogenous effector functions (e.g. immunodeficient rats or mice), or (ii) transplanting mouse cancer cells into which HLA and NY-ESO genes have been transduced into non-human animals with a knockout of the human CD3 gene, and preferably rats or mice. By performing the evaluation using such immunodeficient animals or knockout animals, various assays, immunohistochemical assays and the like can be carried out using the bodies of mice and/or rats. This is preferable for medicaments containing the multispecific molecule of the present invention for preclinical development and other purposes.

In the present invention, "recognition"; i.e. "binding" is binding that is not non-specific adsorption. Whether or not the antibody recognizes, i.e. binds, can be assessed based on, for example, the dissociation constant (KD). A preferable KD value of the antibody and the like of the present invention for HLA/NY-ESO or CD3 is 1× ^10-5 M or less, 5× ^10-6 M or less, 2× ^10-6 M or less, or 1× ^10-6 M or less, their KD value for HLA/NY-ESO is preferably 5× ^10-7 M or less, 2× ^10-7 M or less, 1× ^10-7 M or less, 5× ^10-8 M or less, 2× ^10-8 M or less, 1× ^10-8 M or less, 5× ^10-9 M or less, or 2× ^10-9 M or less, and more preferably 1× ^10-9 M or less. Examples of the anti-HLA/NY-ESO scFv of the present invention having excellent antigen-binding activity include NYA-1143, NYA-2023, NYA-2143, NYA-2044, NYA-2045, NYA-2060, NYA-2061 and NYA-3061, and their KD value for HLA/NY-ESO such as NYA-1143, NYA-2044, NYA-2045 and NYA-2143 is 1× ^10-9 M or less (e.g., Example 4).

In the present invention, antigen-antibody binding can be analyzed or assessed, for example, using a biomolecular interaction analysis system such as SPR or BLI, ELISA or RIA. Binding between an antigen and an antibody expressed on the cell surface can be assessed, for example, using flow cytometry.

Surface plasmon resonance (SPR) analysis method performs reaction kinetic analysis to determine the association rate constant (Ka value) and dissociation rate constant (Kd value), and determines the dissociation constant (KD value), which serves as an indicator of affinity. Examples of devices used for SPR analysis include Biacore™ (GE Healthcare), ProteOn™ (BioRad), SPR-Navi™ (BioNavis), Spreeta™ (Texas Instruments), SPRi-PlexII™ (Horiba Ltd.), and Autolab SPR™ (Metrohm).

The biolayer interferometry (BLI) method analyzes interactions between biomolecules using biolayer interference. An example of an apparatus used for interaction analysis by the BLI method is the Octet system (Pall ForteBio).

The enzyme-linked immunosorbent assay (ELISA) method detects and quantifies a target antigen or antibody contained in a sample solution by capturing the target antigen or antibody using a specific antibody or antigen, respectively, using an enzymatic reaction. The enzyme-labeled antigen or antibody is integrated into a reaction system to determine the enzymatic activity. The enzymatic activity is determined using a substrate that exhibits absorption spectra that change depending on the reaction, and the enzymatic activity is quantified based on the absorption analysis.

Cell-ELISA detects and quantifies the target analyte on the cell surface by capturing the target analyte along with the cell using an enzymatic reaction.

The radioimmunoassay (RIA) method involves labeling an antibody with a radioactive substance and determining the radioactivity of the antibody. In this way, the antibody can be quantified.

In the flow cytometry method, cells are dispersed in a liquid, the liquid is allowed to flow narrowly, and each cell is subjected to optical analysis. An antibody labeled with a fluorescent dye is allowed to bind to a cell surface antigen through an antigen-antibody reaction, and the intensity of fluorescence emitted by the labeled antibody bound to the cell is analyzed to quantify the antigen-binding ability of the antibody.

Examples of the anti-HLA/NY-ESO antibodies of the present invention exhibiting excellent antigen-binding specificity include anti-HLA/NY-ESO scFv such as NYA-0001, NYA-1143, NYA-1163, NYA-2023, NYA-2027, NYA-2035, NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060, NYA-2061, NYA-2143 and NYA-3061 (e.g., Example 6).

3. An antibody that specifically binds to HLA/NY-ESO or its binding fragment.

3-1. Anti-HLA/NY-ESO or its binding fragment

The present invention provides an antibody that recognizes and binds to HLA/NY-ESO or a binding fragment thereof.

As described above, HLA/NY-ESO is a complex containing HLA-A2 and 9-mer NY-ESO peptide (SLLMWITQC: SEQ ID NO:1). NY-ESO peptide is a peptide derived from NY-ESO-1 or LAGE-1, which is an intracellular protein and testicular cancer antigen. HLA/NY-ESO is expressed on the surface of cancer cells.

The anti-HLA/NY-ESO antibody of the present invention and the antigen-binding fragment of the antibody (which may be referred to as "the antibody and the like of the present invention" hereinafter) may be monoclonal or polyclonal antibodies. In the present invention, the isotype of the monoclonal antibody is not particularly limited, and examples include IgG such as IgG1, IgG2, IgG3 and IgG4, IgM, IgA such as IgA1 and IgA2, IgD and Ig. The isotype and subclass of the monoclonal antibody can be determined by, for example, the Ouchterlony method, ELISA or RIA. Examples of the monoclonal antibodies of the present invention include an antibody derived from a non-human animal (non-human antibody), a human antibody, a chimerized antibody (also referred to as "chimeric antibody") and a humanized antibody, wherein a human antibody is preferable. The antibody of the present invention includes a mutant antibody (a "mutant antibody" described below), and, for example, a human antibody includes a mutant human antibody.

Examples of non-human animal antibodies include those derived from vertebrates such as mammals and birds. Examples of mammalian-derived antibodies include those derived from rodents such as mouse antibody and rat antibody. An example of an avian-derived antibody is chicken antibody.

Examples of chimerized antibodies include, but are not limited to, antibodies comprising a variable region derived from a non-human animal antibody linked to a constant region derived from a human antibody (human immunoglobulin).

Examples of humanized antibodies include, but are not limited to, a humanized antibody obtained by grafting a CDR in a variable region of a non-human antibody into a human antibody (a human immunoglobulin variable region), a humanized antibody obtained by grafting, in addition to the CDR, a portion of a framework region sequence of a non-human antibody into a human antibody, and a humanized antibody obtained by replacing 1 or more amino acids derived from a non-human antibody with amino acids derived from a human antibody.

The antibody can be produced by various known methods. For example, the antibody can be produced by a method involving the use of a hybridoma, cell-mediated immunity or genetic recombination. In addition, a human antibody obtained by phage display selected from a human antibody library can be obtained. For example, in the phage display method, a variable region of a human antibody can be expressed as an scFv on the surface of a phage, after which an antigen-binding phage can be selected. The gene of the phage selected by its binding to the antigen can be analyzed so that the DNA sequence encoding the variable region of the human antigen binding to the antigen can be determined. If the DNA sequence of the antigen-binding scFv is elucidated, an expression vector containing such a sequence can be obtained, introduced into a suitable host cell and expressed therein. In this way, a human antibody can be obtained (WO 92/01047, WO 92/20791, WO 93/06213, WO 93/11236, WO 93/19172, WO 95/01438, WO 95/15388, Annu. Rev. Immunol., 1994, 12, 433-455).

The antibody thus obtained with high activity can be used as a lead antibody, and the gene encoding such a lead antibody can be mutated to produce a mutant with higher activity (a “mutant antibody” described below).

A preferred combination of CDRH1-CDRH3 included in the heavy chain of the anti-HLA/NY-ESO antibody of the present invention, or an antigen-binding fragment thereof, is CDRH1 consisting of the amino acid sequence of SEQ ID NO:54 (Fig. 61), CDRH2 consisting of the amino acid sequence of SEQ ID NO:55 (Fig. 61), and CDRH3 consisting of the amino acid sequence of SEQ ID NO:56 (Fig. 61), or an amino acid sequence derived from the amino acid sequence of SEQ ID NO:56 (Fig. 61), in which amino acid 6 is N (Asn). A more preferred CDRH1-CDRH3 combination is comprised in the variable region of the heavy chain of NYA-0001 consisting of the amino acid sequence of SEQ ID NO: 6 (Fig. 13), the variable region of the heavy chain of NYA-0082 consisting of the amino acid sequence of SEQ ID NO: 18 (Fig. 25), the variable region of the heavy chain of NYA-2023 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 27 (Fig. 34), the variable region of the heavy chain of NYA-2027 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 28 (Fig. 35), the variable region of the heavy chain of NYA-1143 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 29 (Fig. 36), the variable region of the heavy chain NYA-1163 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 26 (Fig. 33), the variable region of the heavy chain NYA-2023 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 27 (Fig. 34), the variable region of the heavy chain NYA-2027 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 28 (Fig. 35), the variable region of the heavy chain NYA-2035 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO: 36 (Fig. 43), the variable region of the heavy chain NYA-2044 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:47 (Fig. 54), the variable region of the heavy chain of NYA-2045 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:48 (Fig. 55), the variable region of the heavy chain of NYA-2047 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:50 (Fig. 57), the variable region of the heavy chain of NYA-2048 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:51 (Fig. 58), the variable region of the heavy chain of NYA-2060 consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:52 (Fig. 59), the variable region of the heavy chain of NYA-2061 consisting of the amino acid sequence of amino acids 21-140 amino acid sequence of SEQ ID NO:53 (Fig. 60), or the variable region of the heavy chain of NYA-2143, consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:30 (Fig. 37).

Another example of a CDRH1-CDRH3 combination is included in the variable region of the heavy chain of NYA-3061, consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:156 (Fig. 152).

A preferred CDRL1-CDRL3 combination included in the light chain of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof is CDRL1 consisting of the amino acid sequence of SEQ ID NO: 57 (Fig. 61) or an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 57 (Fig. 61), wherein amino acid 7 is W (Trp) or amino acid 8 is K (Lys), CDRL2 consisting of the amino acid sequence of SEQ ID NO: 58 (Fig. 61), and CDRL3 consisting of the amino acid sequence of SEQ ID NO: 59 (Fig. 61) or an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 59 (Fig. 61), wherein amino acid 2 is A (Ala) or S (Ser).

A more preferred combination of CDRL1-CDRL3 is comprised of the variable light region of NYA-0001 consisting of the amino acid sequence of SEQ ID NO: 8 (Fig. 15), the variable light region of NYA-0082 consisting of the amino acid sequence of SEQ ID NO: 20 (Fig. 27), the variable light region of NYA-1143 consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 29 (Fig. 36), the variable light region of NYA-1163 consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 26 (Fig. 33), the variable light region of NYA-2023 consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 27 (Fig. 34), the variable light region of NYA-2027, consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 28 (Fig. 35), the variable region of the light chain of NYA-2035, consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 36 (Fig. 43), the variable region of the light chain of NYA-2044, consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 47 (Fig. 54), the variable region of the light chain of NYA-2045, consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 48 (Fig. 55), the variable region of the light chain of NYA-2047, consisting of amino acids 156-266 of the amino acid sequence of SEQ ID NO: 50 (Fig. 57), the variable region of the light chain of NYA-2048, consisting of amino acids 156-266 of the amino acid sequence SEQ ID NO:51 (Fig. 58), the variable region of the light chain of NYA-2060 consisting of amino acids 156-266 of the amino acid sequence SEQ ID NO:52 (Fig. 59), the variable region of the light chain of NYA-2061 consisting of amino acids 156-266 of the amino acid sequence SEQ ID NO:53 (Fig. 60), or the variable region of the light chain of NYA-2143 consisting of amino acids 156-266 of the amino acid sequence SEQ ID NO:30 (Fig. 37).

Another example of a CDRL1-CDRL3 combination is included in the variable region of the light chain of NYA-3061, consisting of amino acids 161-271 of the amino acid sequence SEQ ID NO:156 (Fig. 152).

A preferred combination of CDRH1-CDRH3 in the heavy chain and CDRL1-CDRL3 in the light chain of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof is a combination of CDRH1 consisting of the amino acid sequence of SEQ ID NO: 54 (Fig. 61), CDRH2 consisting of the amino acid sequence of SEQ ID NO: 55 (Fig. 61), and CDRH3 consisting of the amino acid sequence of SEQ ID NO: 56 (Fig. 61) or an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 56 (Fig. 61) in which amino acid 6 is N (Asn), and a combination of CDRL1 consisting of the amino acid sequence of SEQ ID NO: 57 (Fig. 61) or an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 57 (Fig. 61) in which amino acid 7 is N (Asn) and/or amino acid 8 is K (Lys), CDRL2 consisting of the amino acid sequence of SEQ ID NO:58 (Fig. 61), and CDRL3 consisting of the amino acid sequence as shown in SEQ ID NO:59 (Fig. 61) or an amino acid sequence derived from the amino acid sequence of SEQ ID NO:59, in which amino acid 2 is A (Ala) or S (Ser). A more preferred combination of CDRH1-CDRH3 and CDRL1-CDRL3 is comprised of the heavy chain variable region and the light chain variable region of NYA-0001 consisting of the amino acid sequences shown in SEQ ID NO:6 and SEQ ID NO:8, the heavy chain variable region and the light chain variable region of NYA-1143 consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence of SEQ ID NO:29 (FIG. 36), the heavy chain variable region and the light chain variable region of NYA-1163 consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence of SEQ ID NO:26 (FIG. 33), the heavy chain variable region and the light chain variable region of NYA-2023 consisting of amino acids 21-140 and amino acids 156-266 amino acid sequence of SEQ ID NO:27 (Fig. 34), the variable region of the heavy chain and the variable region of the light chain of NYA-2027 consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence shown in SEQ ID NO:28 (Fig. 35), the variable region of the heavy chain and the variable region of the light chain of NYA-2035 consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence of SEQ ID NO:36 (Fig. 43), the variable region of the heavy chain and the variable region of the light chain of NYA-2044 consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence of SEQ ID NO:47 (Fig. 54), the variable region of the heavy chain and the variable region of the light chain NYA-2045, consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence SEQ ID NO: 48 (Fig. 55), the variable region of the heavy chain and the variable region of the light chain of NYA-2047, consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence SEQ ID NO: 50 (Fig. 57), the variable region of the heavy chain and the variable region of the light chain of NYA-2048, consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence SEQ ID NO: 51 (Fig. 58), the variable region of the heavy chain and the variable region of the light chain of NYA-2060, consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence SEQ ID NO: 52 (Fig. 59), the variable region of the heavy chain and the variable region of the light chain of NYA-2061, consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence of SEQ ID NO:53 (Fig. 60), or the variable region of the heavy chain and the variable region of the light chain of NYA-2143, consisting of amino acids 21-140 and amino acids 156-266 of the amino acid sequence of SEQ ID NO:30 (Fig. 37).

Another example of a combination of CDRH1-CDRH3 and CDRL1-CDRL3 is included in the variable region of the heavy chain and the variable region of the light chain of NYA-3061, consisting of amino acids 21-140 and amino acids 161-271 of the amino acid sequence SEQ ID NO:156 (Fig. 152).

Preferred examples of the heavy chain variable region of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof include the heavy chain CDRs described above and variable regions including such heavy chain CDRs in appropriate combination. More preferable examples include NYA-0001 heavy chain variable region, NYA-0082 heavy chain variable region, NYA-1143 heavy chain variable region, NYA-1163 heavy chain variable region, NYA-2023 heavy chain variable region, NYA-2027 heavy chain variable region, NYA-2035 heavy chain variable region, NYA-2044 heavy chain variable region, NYA-2045 heavy chain variable region, NYA-2047 heavy chain variable region, NYA-2048 heavy chain variable region, NYA-2060 heavy chain variable region, NYA-2061 heavy chain variable region, NYA-2143 heavy chain variable region and NYA-3061 heavy chain variable region. The amino acid sequence of each heavy chain variable region is as described above.

Preferred examples of the light chain variable region of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof include the light chain CDRs described above and variable regions comprising such light chain CDRs in appropriate combination. More preferable examples include NYA-0001 light chain variable region, NYA-0082 light chain variable region, NYA-1143 light chain variable region, NYA-1163 light chain variable region, NYA-2023 light chain variable region, NYA-2027 light chain variable region, NYA-2035 light chain variable region, NYA-2044 light chain variable region, NYA-2045 light chain variable region, NYA-2047 light chain variable region, NYA-2048 light chain variable region, NYA-2060 light chain variable region, NYA-2061 light chain variable region, NYA-2143 light chain variable region and NYA-3061 light chain variable region. The amino acid sequence of each light chain variable region is as described above.

Preferred examples of the heavy chain variable region and the light chain variable region of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof include the heavy and light chain CDRs described above or CDRs including such CDRs in adequate combination. More preferable examples include the variable region of the heavy chain and the variable region of the light chain of NYA-0001, the variable region of the heavy chain and the variable region of the light chain of NYA-0082, the variable region of the heavy chain and the variable region of the light chain of NYA-1143, the variable region of the heavy chain and the variable region of the light chain of NYA-1163, the variable region of the heavy chain and the variable region of the light chain of NYA-2023, the variable region of the heavy chain and the variable region of the light chain of NYA-2027, the variable region of the heavy chain and the variable region of the light chain of NYA-2035, the variable region of the heavy chain and the variable region of the light chain of NYA-2044, the variable region of the heavy chain and the variable region of the light chain of NYA-2045, the variable region of the heavy chain and the variable region of the the light chain region of NYA-2047, the variable heavy chain region and the variable light chain region of NYA-2048, the variable heavy chain region and the variable light chain region of NYA-2060, the variable heavy chain region and the variable light chain region of NYA-2061, the variable heavy chain region and the variable light chain region of NYA-2143, and the variable heavy chain region and the variable light chain region of NYA-3061.

Preferred examples of the heavy chain of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof include heavy chains including the preferred or more preferred heavy chain variable regions described above. More preferred examples include NYA-0001 heavy chain, NYA-0082 heavy chain, NYA-1143 heavy chain, NYA-1163 heavy chain, NYA-2023 heavy chain, NYA-2027 heavy chain, NYA-2035 heavy chain, NYA-2044 heavy chain, NYA-2045 heavy chain, NYA-2047 heavy chain, NYA-2048 heavy chain, NYA-2060 heavy chain, NYA-2061 heavy chain, NYA-2143 heavy chain and NYA-3061 heavy chain.

Preferred examples of the light chain of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof include light chains including the preferred or more preferred light chain variable regions described above. More preferred examples include NYA-0001 light chain, NYA-0082 light chain, NYA-1143 light chain, NYA-1163 light chain, NYA-2023 light chain, NYA-2027 light chain, NYA-2035 light chain, NYA-2044 light chain, NYA-2045 light chain, NYA-2047 light chain, NYA-2048 light chain, NYA-2060 light chain, NYA-2061 light chain, NYA-2143 light chain and NYA-3061 light chain.

Preferred examples of the heavy chain and light chain of the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof include heavy chains and light chains including the preferred or more preferred heavy chain variable regions and light chain variable regions described above. More preferable examples include NYA-0001 heavy chain and light chain, NYA-1143 heavy chain and light chain, NYA-1163 heavy chain and light chain, NYA-2023 heavy chain and light chain, NYA-2027 heavy chain and light chain, NYA-2035 heavy chain and light chain, NYA-2044 heavy chain and light chain, NYA-2045 heavy chain and light chain, NYA-2047 heavy chain and light chain, NYA-2048 heavy chain and light chain, NYA-2060 heavy chain and light chain, NYA-2061 heavy chain and light chain, NYA-2143 heavy chain and light chain and NYA-3061 heavy chain and light chain.

An antigen-binding fragment of an antibody is a fragment that retains at least the antigen-binding ability beyond the functions of the original antibody or a modified product thereof. Examples of antibody functions include, in general, antigen-binding activity, antigen-regulating activity, antibody-dependent cellular cytotoxicity, and complement-dependent cytotoxicity. Examples of functions of the antibody and the like of the present invention and multispecific molecules containing the antibody and the like of the present invention include T-cell redirection, T-cell activation, and T-cell activation-induced anti-cancer cytotoxicity.

The antigen-binding fragment of the antibody is not particularly limited as long as it is derived from an antibody that retains at least the antigen-binding ability beyond the functions of the original antibody. Examples thereof include, but are not limited to, Fab, Fab', F(ab') ₂ , Fv, single-chain Fv (scFv) comprising a heavy chain Fv ligated to a light chain Fv via an appropriate linker, and a single-domain antibody (sdAb). A molecule comprising a region other than the antigen-binding fragment of the antibody of the present invention, as in the case of scFv comprising a linker portion, is included within the scope of the antigen-binding fragment of the antibody of the present invention.

An antibody protein molecule lacking at least one or more amino acids from the amino terminus and/or carboxyl terminus and retaining some functions of the antibody is included within the scope of an antigen-binding fragment of an antibody. A modified antigen-binding fragment of an antibody is included within the scope of an antibody of the present invention, an antigen-binding fragment thereof, or a modified antibody or fragment (described below).

An embodiment of the antibody of the present invention or antigen-binding fragment thereof is an scFv. The scFv is obtained by ligating the variable region of the heavy chain of an antibody to the variable region of its light chain using a polypeptide linker (Pluckthun A., The Pharmacology of Monoclonal Antibodies 113, Rosenburg and Moore (ed.), Springer Verlag, New York, 269-315, 1994, Nature Biotechnology, 2005, 23, 1126-1136). In addition, a tandem scFv obtained by linking two scFv constructs with a polypeptide linker can be used as a bispecific molecule. In addition, a triabody or the like containing 3 or more scFv constructs can be used as a multispecific molecule.

Preferred examples of the HLA/NY-ESO-specific scFv (also referred to as “anti-HLA/NY-ESO scFv”) include an scFv comprising the CDRH1 to CDRH3 and CDRL1 to CDRL3 described above, more preferred examples thereof include an scFv comprising a heavy chain variable region and a light chain variable region, and further preferred examples thereof include NYA-0001 (amino acids 21 to 266 of the amino acid sequence of SEQ ID NO: 70 (Fig. 71)), NYA-0082 (comprising the amino acid sequence shown in SEQ ID NO: 18 (Fig. 25) and the amino acid sequence shown in SEQ ID NO: 20 (Fig. 27)), NYA-1143 (amino acids 21 to 266 of the amino acid sequence shown in SEQ ID NO: 29 (Fig. 36)), NYA-1163 (amino acids 21-266 of the amino acid sequence SEQ ID NO:26 (Fig. 33)), NYA-2023 (amino acids 21-266 of the amino acid sequence SEQ ID NO:27 (Fig. 34)), NYA-2027 (amino acids 21-266 of the amino acid sequence SEQ ID NO:28 (Fig. 35), NYA-2035 (amino acids 21-266 of the amino acid sequence SEQ ID NO:36 (Fig. 43)), NYA-2044 (amino acids 21-266 of the amino acid sequence SEQ ID NO:47 (Fig. 54)), NYA-2045 (amino acids 21-266 of the amino acid sequence SEQ ID NO:48 (Fig. 55)), NYA-2047 (amino acids 21-266 of the amino acid sequence SEQ ID NO:50 (Fig. 57)), NYA-2048 (amino acids 21-266 of the amino acid sequence SEQ ID NO:51 (Fig. 58)), NYA-2060 (amino acids 21-266 of the amino acid sequence SEQ ID NO:52 (Fig. 59)), NYA-2061 (amino acids 21-266 of the amino acid sequence SEQ ID NO:53 (Fig. 60)), and NYA-2143 (amino acids 21-266 of the amino acid sequence SEQ ID NO:30 (Fig. 37)). Another example is NYA-3061 (amino acids 21-271 of the amino acid sequence SEQ ID NO:156 (Fig. 152)).

A preferred embodiment of the anti-HLA/NY-ESO scFv is an scFv comprising a FLAG-His tag fused to its carboxyl terminus (may be referred to simply as the “tag adduct”). Examples of preferred tag adducts include NYA-0001 tag adduct (amino acids 20-292 of SEQ ID NO:70 (FIG. 71)), NYA-1143 tag adduct (amino acids 20-292 of SEQ ID NO:29 (FIG. 36)), NYA-1163 tag adduct (amino acids 20-292 of SEQ ID NO:26 (FIG. 33)), NYA-2023 tag adduct (amino acids 20-292 of SEQ ID NO:27 (FIG. 34) ), NYA-2027 tag adduct (amino acids 20-292 of SEQ ID NO:28 (FIG. 35)), NYA-2035 tag adduct (amino acids 20-292 of SEQ ID NO:36 (Fig. 43)), NYA-2044 tag adduct (amino acids 20-292 of SEQ ID NO:47 (Fig. 54)), NYA-2045 tag adduct (amino acids 20-292 of SEQ ID NO:48 (Fig. 55)), NYA-2047 tag adduct (amino acids 20-292 of SEQ ID NO:50 (Fig. 57)), NYA-2048 tag adduct (amino acids 20-292 of SEQ ID NO:51 (Fig. 58)), NYA-2060 tag adduct (amino acids 20-292 of SEQ ID NO:52 (Fig. 59)), NYA-2061 tag adduct (amino acids 20-292 of SEQ ID NO:53 (Fig. 60)) and an adduct of the NYA-2143 tag (amino acids 20-292 of SEQ ID NO:30 (Fig. 37)).

Of these, NYA-2023 and its label adduct, NYA-2047 and its label adduct, NYA-2048 and its label adduct, NYA-2060 and its label adduct, and NYA-2061 and its label adduct exhibit excellent biological activity, physical properties and the like in Fc-added bispecific molecules, and are therefore more preferable.

When the anti-HLA/NY-ESO scFv and its tag adduct are to be expressed in a host cell, a signal peptide may be added to its amino terminus. Examples of amino acid sequences of the anti-HLA/NY-ESO scFv tag adducts with the addition of a signal peptide include the amino acid sequences shown in SEQ ID NOs: 70, 29, 26-28, 36, 47, 48, 50-53, and 30 (Fig. 71, 36, 33-35, 43, 54, 55, 57-60, and 37).

scFv can be produced by a phage display method in which the variable region of an antibody is expressed as a single-chain antibody (scFv) on the surface of a phage, and then an antigen-binding phage is selected (Nature Biotechnology, 2005, 23, (9), pp. 1105-1116). The gene of the phage selected by its binding to the antigen can be analyzed to determine the DNA sequence encoding the variable region of the human antigen that binds to the antigen. If the DNA sequence of the antigen-binding scFv is elucidated, an expression vector containing such a sequence can be prepared, introduced into a suitable host cell, and expressed therein. In this way, a human antibody can be obtained (WO 92/01047, WO 92/20791, WO 93/06213, WO 93/11236, WO 93/19172, WO 95/01438, WO 95/15388, Annu. Rev. Immunol., 1994, 12, pp. 433-455, Nature Biotechnology, 2005, 23 (9), pp. 1105-1116).

The antibody of the present invention may comprise one variable region of the heavy chain and may not comprise a light chain sequence. Such an antibody is called a single-domain antibody (sdAb) or nanobody and retains antigen-binding ability (Muyldemans S. et al., Protein Eng., 1994, 7 (9), 1129-35, Hamers-Casterman C. et al., Nature, 1993, 363 (6428), 446-448). Such antibodies are included within the scope of the antigen-binding fragment of the antibody of the present invention.

The present invention also includes a single-chain immunoglobulin comprising a full-length heavy chain sequence ligated to a light chain sequence of an antibody using an appropriate linker (Lee, H-S, et al., Molecular Immunology, 1999, 36, 61-71; Shirrmann, T. et al., mAbs, 2010, 2 (1), 1-4). Such a single-chain immunoglobulin may be dimerized so that it may retain a structure and activity similar to that of an antibody that is tetrameric in nature. The anti-HLA/NY-ESO antibody of the present invention may be a single-chain immunoglobulin.

In the scFv of the present invention, the variable region of the heavy chain may form a disulfide bond with the variable region of the light chain.

The anti-HLA/NY-ESO antibody of the present invention may be composed of components derived from a plurality of different antibodies, as long as it binds to HLA/NY-ESO. Examples thereof include those derived by substituting heavy chains and/or light chains among a plurality of different antibodies, substituting the entire sequences of the heavy chains and/or light chains, substituting by selectively replacing any variable region or constant region, and those derived by selectively replacing part or all of the CDRs. In the chimerized antibody, the variable region of the heavy chain and the variable region of the light chain may be derived from another anti-HLA/NY-ESO antibody of the present invention. In the variable regions of the heavy chain and the light chain of the humanized antibody, CDR1 of the heavy chain to CDR3 of the heavy chain and CDR1 of the light chain to CDR3 of the light chain may be derived from two or more types of the anti-HLA/NY-ESO antibodies according to the present invention. In the variable regions of the heavy chain and light chain of a human antibody, the combination of heavy chain CDR1 - heavy chain CDR3 and light chain CDR1 - light chain CDR3 may be obtained from two or more types of the anti-HLA/NY-ESO antibodies of the present invention.

The anti-HLA/NY-ESO antibody of the present invention includes an antibody that comprises an amino acid sequence encoded by a nucleotide sequence included in a polynucleotide that hybridizes under stringent conditions to a complementary chain of a polynucleotide containing a nucleotide sequence encoding an amino acid sequence included in the anti-HLA/NY-ESO antibody of the present invention and binds to HLA/NY-ESO.

It may be an antibody comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to an amino acid sequence comprised in the variable region of the heavy chain of the anti-HLA/NY-ESO antibody of the present invention (preferably the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:27, the amino acid sequence shown in SEQ ID NO:38, the amino acid sequence shown in SEQ ID NO:39, or the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:160, SEQ ID NO:197 or SEQ ID NO:198) and/or an amino acid sequence comprised in the variable region of the light chain thereof (preferably the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:27, the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:52, the amino acid sequence shown in SEQ ID NO:40, the amino acid sequence of amino acids 161-271 of the amino acid sequence of SEQ ID NO:160, or the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:197 or SEQ ID NO:198) or an antigen-binding fragment thereof.

When the position and length of the light chain variable region are determined according to a definition that differs from IMGT (e.g., Kabat, Chothia, AbM, Contact), one or more amino acids, such as arginine or glycine, may be further included at the carboxyl terminus of the amino acid sequence of the light chain variable region determined according to the IMGT definition. Such an antibody or a binding fragment thereof comprising such a light chain variable region is included within the scope of the antibody of the present invention or a binding fragment thereof.

The antibody and the like of the present invention can be obtained by introducing a mutation into the binding fragment of the anti-HLA/NY-ESO antibody of the present invention and optimizing the ability to bind to HLA/NY-ESO, and in particular to human and/or cynomolgus monkey HLA/NY-ESO. Specific examples of methods for introducing a mutation include random mutagenesis using error-prone PCR, site-directed introduction of amino acids using NHK libraries, site-directed mutagenesis using structural information, and any combination thereof.

3-2. Mutant anti-HLA/NY-ESO antibody (mutant antibody)

The mutant antibody of the anti-HLA/NY-ESO antibody of the present invention can be preferably provided with, for example, reduced susceptibility to protein degradation or oxidation, maintained or improved biological activity or functions, suppression of decrease or change in such activity or functions, improved or adjustable antigen-binding ability, physicochemical properties or functional properties. It is known that a protein changes its functions or activity by alternating a specific amino acid side chain on its surface, and examples include deamidation of the asparagine side chain and isomerization of the aspartic acid side chain. An antibody obtained by replacing a specific amino acid with another amino acid to prevent the change in the amino acid side chain is included in the scope of the mutant antibody of the present invention.

An example of a mutant antibody of the present invention is an antibody comprising an amino acid sequence derived from the amino acid sequence of the original antibody by a conservative amino acid substitution. The conservative amino acid substitution occurs within an amino acid group associated with the side chain of the amino acid.

Preferred amino acid groups are as follows: acidic group: aspartic acid and glutamic acid; basic group: lysine, arginine and histidine; non-polar group: alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine and tryptophan; and uncharged polar family: glycine, asparagine, glutamine, cysteine, serine, threonine and tyrosine. Other preferred amino acid groups are as follows: aliphatic hydroxyl group: serine and threonine; amide-containing group: asparagine and glutamine; aliphatic group: alanine, valine, leucine and isoleucine; and aromatic group: phenylalanine, tryptophan and tyrosine. In such a mutant antibody, the amino acid substitution is preferably carried out while refraining from reducing the antigen-binding activity of the original antibody.

The anti-HLA/NY-ESO antibody of the present invention, its antigen-binding fragment, its mutant (mutant antibody or antigen-binding fragment thereof) or molecule of the present invention includes: a mutant antibody comprising an amino acid sequence derived from the amino acid sequence of an antibody of the present invention such as NYA-2023 by a conservative amino acid substitution and/or other mutation and binding to HLA/NY-ESO, its antigen-binding fragment, and molecules comprising them; a chimerized antibody, a humanized antibody or a human antibody comprising a CDR of an amino acid sequence derived from the amino acid sequence of any of CDRH1-CDRH3 and CDRL1-CDRL3 derived from an antibody of the present invention including NYA-2023 by a conservative amino acid substitution and/or other mutation and binding to HLA/NY-ESO, its antigen-binding fragment or a molecule comprising them.

3-3. Binding fragment of anti-HLA/NY-ESO antibody

In one aspect of the present invention, an antigen-binding fragment of an anti-HLA/NY-ESO antibody of the present invention is provided (hereinafter referred to simply as a "binding fragment"). The binding fragments of the anti-HLA/NY-ESO antibody of the present invention include binding fragments of a chimerized antibody, a humanized antibody, and a human antibody. The binding fragment of an antibody is a fragment that retains at least the antigen-binding ability of the functions of the original antibody or a modified product thereof. In general, examples of such functions of antibodies include antigen-binding activity, antigen activity regulation activity (e.g., agonist activity), antigen internalization activity into a cell, and activity of inhibiting or stimulating interactions between an antigen and a substance interacting with it.

The binding fragment of the antibody is not particularly limited as long as it is a fragment of a fragment retaining at least the antigen-binding activity of the original antibody. Examples of such a binding fragment of an antibody include, but are not limited to, Fab, Fab', F(ab') ₂ , single-chain Fab (scFab) in which the carboxyl terminus of the light chain of a Fab is ligated to the amino terminus of the heavy chain of a Fab via an appropriate linker, Fv, single-chain Fv (scFv) comprising a heavy chain Fv ligated to a light chain Fv via an appropriate linker, and a single-domain antibody (sdAb) with one variable region of the heavy chain but without the light chain sequence, which is also called a nanobody (Muyldemans S. et al., Protein Eng., 1994, 7 (9), 1129-35, Hamers-Casterman C. et al., Nature, 1993, 363 (6428), 446-448). A molecule containing regions other than the binding fragment of the antibody of the present invention, such as an scFab or scFv containing a linker region, is included within the scope of the binding fragment of the antibody of the present invention.

3-4. Modified anti-HLA/NY-ESO antibody, its modified binding fragment or their complex

The present invention provides a modified antibody or a modified binding fragment thereof. The modified antibody of the present invention or a modified binding fragment thereof is subjected to chemical or biological modification. Examples of chemical modification include linking a chemical moiety to an amino acid backbone and chemical modification of N-linked or O-linked carbohydrate chains. Examples of biological modification include post-translational modification (e.g., adding a sugar chain to an N- or O-linkage, treating the amino-terminal or carboxy-terminal region, deamidation, isomerization of aspartic acid, and oxidation of methionine) and adding methionine to the amino terminus by expression in a prokaryotic host cell. In addition, labels that allow detection or isolation of the antibody or antigen of the present invention, such as an enzyme label, a fluorescent label, and an affinity label, are included in the scope of the modified antibody or antigen as described above. The modified antibody of the present invention or its binding fragment as described above can be used to improve the stability and persistence in blood of the original antibody of the present invention or its binding fragment, reduce antigenicity, detect or isolate the antibody or antigen, and other purposes.

Examples of chemical moieties included in a chemically modified antibody or fragment include water-soluble polymers such as polyethylene glycol (PEG), ethylene glycol/propylene glycol polymer, carboxymethylcellulose, dextran, and polyvinyl alcohol.

Examples of biological modification include enzyme treatment, cell treatment, addition of other peptides such as tags through genetic recombination, and the use of host cells expressing enzymes modified with endogenous or exogenous sugar chains.

Such modification may be provided at any desired position in the antibody or binding fragment thereof, and the same or two or more different types of modification may be provided at one or more positions.

However, deletion of such a heavy chain sequence or modification of the heavy chain or light chain sequence does not significantly affect the antigen-binding capacity and effector functions of the antibody (e.g., complement activation or antibody-dependent cytotoxicity).

Accordingly, the present invention encompasses an antibody subjected to such a deletion or modification. Examples include a deletion mutant lacking 1 or 2 amino acids at the carboxyl terminus of the heavy chain (Journal of Chromatography A; 705; 129-134, 1995), a deletion mutant lacking 2 amino acids (glycine and lysine) at the carboxyl terminus of the heavy chain and subjected to amidation of proline at the carboxyl terminus (Analytical Biochemistry, 360: 75-83, 2007), and an antibody obtained by pyroglutamylation of the amino terminal glutamine or glutamic acid of the heavy or light chain (WO 2013/147153) (collectively referred to as "deletion mutants"). As long as the antigen-binding ability and effector functions are maintained, the antibody of the present invention lacking the end of the heavy chain and the carboxyl terminus of the light chain is not limited to the deletion mutants described above. When the antibody of the present invention comprises 2 or more chains (e.g., heavy chains), any one of such 2 or more chains (e.g., heavy chains) or both may be a full-length heavy chain or a heavy chain selected from the group consisting of the deletion mutants described above. Although the quantitative ratio or the numerical ratio of the deletion mutant molecules will be affected by the type and culture conditions of the cultured mammalian cells producing the antibody of the present invention, the essential components of the antibody of the present invention may be both of the 2 heavy chains lacking the amino acid residue at the carboxyl terminus.

In addition, an antibody of the present invention or an antigen-binding fragment thereof (including those included in the molecule, multispecific molecule and bispecific molecule of the present invention) comprising one to several amino acids derived from an expression vector and/or or a signal sequence added to the amino terminus and/or carboxy terminus (and partially or completely modified as described above) or the like are included within the scope of the modified antibody of the present invention or a modified antigen-binding fragment thereof, as long as the antigen-binding activity of interest is maintained. A molecule comprising such a modified antibody or a modified antigen-binding fragment thereof is included within the scope of the molecule of the present invention.

In the present invention, "an antibody or a binding fragment thereof" includes "a modified antibody or a modified antigen-binding fragment thereof". In addition, "an antibody or an antigen-binding fragment thereof" included in the molecule, multispecific molecule and bispecific molecule of the present invention include "a modified antibody or a modified antigen-binding fragment thereof".

Antibody-dependent cellular cytotoxicity can be enhanced by regulating (glycosylation, afucosylation, and the like) the modification of the sugar chain associated with the antibody of the present invention. Known methods for regulating the modification of the sugar chain of an antibody are described, for example, in WO 99/54342, WO 00/61739, and WO 02/31140, although the methods are not limited thereto.

The present invention encompasses a complex of an antibody and other molecules ligated to each other by a linker (i.e., an immunoconjugate). An example of an antibody-drug complex comprising an antibody bound to a radioactive substance or a compound having pharmacological activity is an antibody-drug conjugate (ADC) (Methods Mol. Biol., 2013, 1045: 1-27; Nature Biotechnology, 2005, 23, pp. 1137-1146).

Furthermore, the present invention encompasses a complex comprising such an antibody linked to another functional polypeptide. An example of such an antibody-peptide complex is a complex of an antibody with an albumin-linked polypeptide (Protein Eng. Des. Sel., 2012, (2): 81-8).

The modified antibody, the antibody with adjustable modification of the sugar chain and the complex described above are included within the scope of the antibody of the present invention, and the binding fragments of the modified antibody, the antibody with adjustable modification of the sugar chain and the complex are within the scope of the binding fragment of the antibody of the present invention.

4. Method for obtaining antibodies

The antibody of the present invention can be produced in a cell as a recombinant antibody, for example, by inserting DNA encoding a heavy chain variable region or DNA encoding a light chain variable region into an expression vector, transforming a host cell for expression with the vector, and culturing the host cell.

With respect to DNA encoding antibodies, DNA encoding a heavy chain can be obtained by ligating DNA encoding a variable region of a heavy chain with DNA encoding a constant region of a heavy chain, and DNA encoding a light chain can be obtained by ligating DNA encoding a variable region of a light chain with DNA encoding a constant region of a light chain.

The anti-HLA/NY-ESO antibody of the present invention can be obtained by inserting the DNA encoding the heavy chain and the DNA encoding the light chain mentioned above into an expression vector, transforming a host cell with the vector, and culturing the host cell. In such a case, the DNA encoding the heavy chain and the DNA encoding the light chain mentioned above can be introduced into the same expression vector, and the host cell can be transformed with the vector. Alternatively, the DNA encoding the heavy chain and the DNA encoding the light chain can be introduced into separate vectors, and the host cell can be transformed with the two vectors. In this case, the DNAs encoding the variable region of the heavy chain and the variable region of the light chain can be introduced into a vector into which the DNA encoding the constant region of the heavy chain and the DNA encoding the constant region of the light chain have been introduced beforehand. In addition, the vector may contain DNA encoding a signal peptide that promotes secretion of the antibody from the host cell. In this case, the DNA encoding the signal peptide and the DNA encoding the antibody are ligated in frame beforehand. The antibody can be obtained as a mature protein by removing the signal peptide after antibody formation.

In this case, DNA encoding a variable region of a heavy chain, DNA encoding a variable region of a light chain, DNA containing DNA encoding a variable region of a heavy chain ligated to DNA encoding a constant region of a heavy chain, or DNA containing DNA encoding a variable region of a light chain ligated to DNA encoding a constant region of a light chain, may be operably ligated to an element such as a promoter, an enhancer, or a polyadenylation signal. When DNA is "operably ligated" as used herein, the DNA is ligated to an element such that the element can perform its functions.

The expression vector is not particularly limited as long as it can be replicated in an animal cell, a bacterial cell, a yeast cell or other host, and examples thereof include known plasmids and phages. Examples of a vector used to create an expression vector include pcDNA™ (Thermo Fisher Scientific), Flexi® vector (Promega), pUC19, pUEX2 (Amersham), pGEX-4T, pKK233-2 (Pharmacia) and pMAMneo (Clontech). As host cells, prokaryotic cells such as Escherichia coli and Bacillus subtilis and eukaryotic cells such as yeast and animal cells can be used, where the use of eukaryotic cells is preferable. Examples of animal cells include human embryonic kidney cell line HEK293 and Chinese hamster ovary (CHO) cells. It is sufficient to introduce the expression vector into a host cell by a known method to transform the host cell. Examples of the methods include an electroporation method, a calcium phosphate precipitation method, and a DEAE-dextran transfection method. The obtained antibody can be purified by conventional protein isolation or purification methods. For example, affinity chromatography or other chromatography methods, filtration, ultrafiltration, salting out, dialysis, and the like can be appropriately selected and combined.

5. Molecule that binds to HLA/NY-ESO

The molecule of the present invention comprises the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof. The molecule of the present invention is preferably a multispecific molecule comprising 2 or more antigen-binding sites. In particular, the molecule of the present invention can bind to 2 or more different epitopes on the molecule or to different epitopes on 2 or more different molecules, and such a molecule comprises a plurality of different antigen-binding fragments. Examples of such a multispecific molecule include, but are not limited to, a multispecific molecule of the IgG type and a multispecific molecule comprising two or more types of variable regions, for example, antibody fragments such as tandem scFv (taFv), single-chain diabody, diabody and triabody, as well as an antibody fragment obtained by covalent or non-covalent binding. The multispecific molecule may comprise an Fc.

The multispecific molecule of the present invention may comprise, in addition to the anti-HLA/NY-ESO antibody of the present invention or an antigen-binding fragment thereof, one or two or more types of antibodies or antigen-binding fragments thereof. Examples of such antigen-binding fragments of an antibody include Fab, F(ab)', Fv, scFv and sdAb.

A preferred multispecific molecule of the present invention further comprises an anti-CD3 antibody or antigen-binding fragment thereof and also specifically binds to CD3.

The anti-CD3 antibody or antigen-binding fragment thereof included in the multispecific molecule of the present invention is not particularly limited, provided that it is a human CD3-binding antibody or binding fragment thereof and it preferably binds to CD3 of a non-human primate such as a cynomolgus macaque. A more preferred example of an anti-CD3 antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof comprising a heavy chain variable region CDRH1 consisting of the amino acid sequence of SEQ ID NO: 141, a heavy chain variable region CDRH2 consisting of the amino acid sequence of SEQ ID NO: 142, a heavy chain variable region CDRH3 consisting of the amino acid sequence of SEQ ID NO: 143, a light chain variable region CDRL1 consisting of the amino acid sequence of SEQ ID NO: 144, a light chain variable region CDRL2 consisting of the amino acid sequence of SEQ ID NO: 145, and a light chain variable region CDRL3 consisting of the amino acid sequence of SEQ ID NO: 146 (Fig. 142).

An example of a more preferred antibody or antigen-binding fragment thereof comprising CDRH1-CDRH3 and CDRL1-CDRL3 is an antibody or antigen-binding fragment thereof comprising a C3E-7034 heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO: 136 (Fig. 137), a C3E-7036 heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO: 137 (Fig. 138), a C3E-7085 heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of SEQ ID NO: 138 (Fig. 139), a C3E-7088 heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:139 (Fig. 140), a C3E-7093 heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:140 (Fig. 141), a C3E-7096 heavy chain variable region consisting of the amino acid sequence of amino acids 272-389 of the amino acid sequence of SEQ ID NO:155 (Fig. 151), a C3E-7096 heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:156 (Fig. 152), and a C3E-7097 heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:157 (Fig. 153).

An example of a more preferred antibody or antigen-binding fragment thereof comprising CDRH1-CDRH3 and CDRL1-CDRL3 is an antibody or antigen-binding fragment thereof comprising a C3E-7034 light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence of SEQ ID NO: 136 (Fig. 137), a C3E-7036 light chain variable region consisting of the amino acid sequence of amino acids 135-241 of the amino acid sequence of SEQ ID NO: 137 (Fig. 138), a C3E-7085 light chain variable region consisting of the amino acid sequence of amino acids 135-241 of the amino acid sequence of SEQ ID NO: 138 (Fig. 139), a C3E-7088 light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:139 (Fig. 140), a light chain variable region C3E-7093 consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:140 (Fig. 141), a light chain variable region C3E-7096 consisting of the amino acid sequence of amino acids 405-511 of the amino acid sequence SEQ ID NO:155 (Fig. 151), a light chain variable region C3E-7096 consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence SEQ ID NO:156 (Fig. 152), and a light chain variable region C3E-7097 consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence SEQ ID NO:157 (Fig. 153).

An example of a more preferred antibody or antigen-binding fragment thereof comprising CDRH1-CDRH3 and CDRL1-CDRL3 is an antibody or antigen-binding fragment thereof comprising a combination of the variable region of the heavy chain and the variable region of the light chain of C3E-7034 consisting of amino acids 2-119 and amino acids 135-243 of SEQ ID NO: 136 (Fig. 137), a combination of the variable region of the heavy chain and the variable region of the light chain of C3E-7036 consisting of amino acids 2-119 and amino acids 135-241 of SEQ ID NO: 137 (Fig. 138), a combination of the variable region of the heavy chain and the variable region of the light chain of C3E-7078 consisting of amino acids 2-119 and amino acids 135-243 of SEQ ID NO: 147 (Fig. 143), a combination variable region of the heavy chain and variable region of the light chain C3E-7085, consisting of amino acids 2-119 and amino acids 135-241 of SEQ ID NO: 138 (Fig. 139), a combination of the variable region of the heavy chain and variable region of the light chain C3E-7088, consisting of amino acids 2-119 and amino acids 135-243 of SEQ ID NO: 139 (Fig. 140), a combination of the variable region of the heavy chain and variable region of the light chain C3E-7093, consisting of amino acids 2-119 and amino acids 135-243 of SEQ ID NO: 140 (Fig. 141), a combination of the variable region of the heavy chain and variable region of the light chain C3E-7096, consisting of amino acids 272-389 and amino acids 405-511 of SEQ ID NO:155 (Fig. 151), a combination of the variable region of the heavy chain and the variable region of the light chain C3E-7096, consisting of amino acids 277-394 and amino acids 410-516 of SEQ ID NO:156 (Fig. 152), and a combination of the variable region of the heavy chain and the variable region of the light chain C3E-7097, consisting of amino acids 277-394 and amino acids 410-516 of SEQ ID NO:157 (Fig. 153).

In addition, examples of a more preferable antibody or antigen-binding fragment thereof comprising CDRH1 to CDRH3 and CDRL1 to CDRL3 include scFv C3E-7034 consisting of an amino acid sequence of amino acids 2 to 243 of the amino acid sequence of SEQ ID NO: 136 (Fig. 137), scFv C3E-7036 consisting of an amino acid sequence of amino acids 2 to 241 of the amino acid sequence of SEQ ID NO: 137 (Fig. 138), scFv C3E-7078 consisting of an amino acid sequence of amino acids 2 to 243 of the amino acid sequence of SEQ ID NO: 147 (Fig. 143), scFv C3E-7085 consisting of an amino acid sequence of amino acids 2 to 241 of the amino acid sequence of SEQ ID NO: 138 (Fig. 139), scFv C3E-7088 consisting of amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO: 139 (Fig. 140), scFv C3E-7093 consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO: 140 (Fig. 141), and an antibody or binding fragment thereof comprising any of such scFv constructs. A preferred embodiment of a CD3-specific scFv (which may be referred to as an "anti-CD3 scFv") comprises an scFv comprising a FLAG-His tag added to its carboxyl terminus (may simply be referred to as a "tag adduct"). Examples of preferred label adducts include C3E-7034 (SEQ ID NO:136; Fig. 137), C3E-7036 (SEQ ID NO:137; Fig. 138), C3E-7085 (SEQ ID NO:138; Fig. 139), C3E-7088 (SEQ ID NO:139; Fig. 140) and C3E-7093 (SEQ ID NO:140; Fig. 141), wherein C3E-7085 is more preferred.

A preferred example of a multispecific molecule according to the present invention is a bispecific molecule. A "bispecific" molecule is capable of binding to two different epitopes on one molecule or to different epitopes on two molecules and includes an antibody or antigen-binding fragment having such bispecificity. The bispecific molecule according to the present invention binds to HLA/NY-ESO and additionally binds to CD3.

Examples of the bispecific molecule of the present invention include those having the structures (formats) described below.

In a dual-type scFv bispecific molecule, each of the two types of scFv constructs binding to different epitopes may bind to one Fc of the dimer with a linker or may directly bind to it without a linker. Alternatively, each of the two types of scFv constructs binding to different epitopes is allowed to bind to CH and CL via linkers and is further allowed to bind to one Fc of the dimer via linkers. In such a bispecific molecule, an Fc containing a heterodimer-forming mutation downstream of one scFv is heterologously associated with another Fc containing a heterodimer-forming mutation downstream of the other scFv. A dual-type scFv bispecific molecule is referred to as a dual-type bispecific molecule or simply as a dual-type molecule (Fig. 6A(b)).

In the present invention, for example, a dual-type bispecific molecule consisting of an anti-HLA-A2/NY-ESO scFv and an anti-CD3 scFv can be used.

Alternatively, the bispecific molecule of the present invention may comprise a Fab and an scFv binding to different epitopes, the Fab of the first antibody may be linked to one Fc of the dimer, and the scFv of the second antibody may be linked to another Fc via linkers. In such a bispecific molecule, the Fc comprising a heterodimer-forming mutation downstream of the Fab is heterologously linked to the Fc comprising a heterodimer-forming mutation downstream of the scFv. Such a bispecific molecule is called a "hybrid-type bispecific molecule" or "hybrid-type" (Fig. 6A(a)). In the present invention, for example, a hybrid-type consisting of an anti-HLA-A2/NY-ESO Fab and an anti-CD3 scFv can be used.

In addition, the bispecific molecule may allow the Fab of the first antibody and the scFv of the second antibody to be attached to a single Fc dimer with linkers. In such a case, the Fab may be allowed to attach to the Fc and the scFv may be allowed to attach to such Fab. Alternatively, the scFv may be allowed to attach to the Fc and the Fab may be allowed to attach to such scFv. Preferably, the Fab may be attached to the Fc and the scFv may be attached to such Fab. The scFv may be attached to the variable region of the Fab via a linker. In such a bispecific molecule, the Fc containing the mutation that forms the heterodimer is linked to a site downstream of the site where the scFv is ligated to the Fab. Such a bispecific molecule is called a scFv-Fab-Fc-type heterodimer bispecific molecule or scFv-Fab-Fc-type heterodimer (Fig. 6A(c)).

In the present invention, for example, an Fc-type scFv-Fab heterodimer consisting of anti-CD3 scFv and anti-HLA-A2/NY-ESO Fab can be used.

In addition, taFv (Fig. 3(c)) comprising scFv constructs of two types (a first antibody and a second antibody) ligated to each other by a linker can be attached to one Fc dimer with a linker or directly without a linker. Such a bispecific molecule is called a taFv-Fc-type heterodimer bispecific molecule or taFv-Fc-type heterodimer (Fig. 3(d)). In such a bispecific molecule, an Fc containing a heterodimer-forming mutation is heterologously linked to another Fc containing a heterodimer-forming mutation at a site downstream of taFv. The order of ligation of the first antibody and the second antibody to taFv is not limited. The previously mentioned bispecific molecule is referred to as taFv-Fc type heterodimer, but when the ligation order of the first antibody and the second antibody to taFv is inverted, the bispecific molecule is referred to as taFv(inverted)-Fc type heterodimer (taFv(inverted)-Fc type).

Fig. 6A(a) shows the structure of the hybrid-type bispecific molecule, Fig. 6A(b) shows the structure of the dual-type bispecific molecule, and Fig. 6A(c) shows the structure of the scFv-Fab-Fc-type heterodimer bispecific molecule. Fig. 3(a) shows the structure of scFv, Fig. 3(b) shows the structure of Fab, Fig. 3(c) shows the structure of taFv, Fig. 3(d) shows the structure of the taFv-Fc-type heterodimer bispecific molecule, and Fig. 3(e) shows the structure of the taFv-Fab-Fc-type heterodimer bispecific molecule. Fig. 6B(a) shows the structure of the taFv-Fc-type heterodimer bispecific molecule (same as in Fig. 3(d)), and Fig. 6B (b) shows the structure of the bispecific taFv (inverted)-Fc type heterodimer molecule, Fig. 6B (c) shows the structure of the first polypeptide included in the bispecific taFv (inverted)-Fc type heterodimer molecule, and Fig. 6 (d) shows the structure of the second polypeptide included in the bispecific taFv (inverted)-Fc type heterodimer molecule.

In the bispecific molecule of the present invention, a plurality of polypeptides are associated.

In the present invention, a taFv consisting of, for example, an anti-HLA-A2/NY-ESO scFv and an anti-CD3 scFv can be used. The Fc type bispecific taFv heterodimer molecule preferably comprises (a) a first polypeptide comprising an scFv that specifically binds to HLA/NY-ESO, an scFv that specifically binds to CD3, and an Fc region (i) of an immunoglobulin in this order from the N terminus to the C terminus, and a second polypeptide comprising a hinge region and an Fc region (ii) of an immunoglobulin. More preferably, it comprises (b) the first polypeptide linked to the second polypeptide in the Fc region (i) and the Fc region (ii). The Fc regions of the first polypeptide and the second polypeptide may comprise a mutation that forms a heterodimer. Fig. 3(d) shows an example of the Fc type bispecific taFv heterodimer molecule. As shown in Fig. 3(d), the Fc region (i) of the first polypeptide binds to the Fc region (ii) of the second polypeptide (solid), and thus the first polypeptide binds to the second polypeptide. Fig. 3(f) shows the first polypeptide, and Fig. 3(g) shows the second polypeptide. In Fig. 3(d), for example, the scFv shown blank is an anti-HLA-A2/NY-ESO scFv, and the scFv indicated with a positive slope is an anti-CD3 scFv.

The first polypeptide included in the more preferred taFv-Fc type heterodimer bispecific molecule of the present invention comprises an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:88, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:89, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:90, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:91, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:92, an amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:93, an amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:94, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:95, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:96, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:86, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:149, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:150, the amino acid sequence of amino acids 20-511 of the amino acid sequence of SEQ ID NO:155, the amino acid sequence of amino acids 20-516 of the amino acid sequence of SEQ ID NO:156, or the amino acid sequence of amino acids 20-516 of the amino acid sequence of SEQ ID NO:157. The first polypeptide included in another preferred taFv-Fc type heterodimer bispecific molecule comprises an amino acid sequence of amino acids 529-745 of the amino acid sequence of SEQ ID NO:85, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 86, 149, 150 or 155, or an amino acid sequence of amino acids 534-750 of the amino acid sequence of SEQ ID NO:156 or 157. The first polypeptide included in an even more preferred taFv-Fc type heterodimer bispecific molecule consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 21-745 amino acid sequence of SEQ ID NO:88, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:89, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:90, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:91, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:92, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:93, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:94, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:95, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:96, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:86, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:149, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:150, or the amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:155. Alternatively, the first polypeptide consists of the amino acid sequence of amino acids 20-750 of the amino acid sequence of SEQ ID NO:156, or the amino acid sequence of amino acids 20-750 of the amino acid sequence of SEQ ID NO:157.

The second polypeptide included in the preferred bispecific taFv-Fc type heterodimer molecule of the present invention comprises a hinge region and a mutant Fc derived from a human antibody. The second polypeptide included in the more preferred bispecific TaFv-Fc type heterodimer molecule comprises an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84.

Of these, NYF-0016, wherein a first polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:85, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0022, wherein the first polypeptide consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:87, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0023, wherein the first polypeptide consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:88, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0027, wherein the first a polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:89 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0035, wherein a first polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:90 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0044, wherein the first polypeptide consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:91 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0045, wherein the first polypeptide consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of sequence of SEQ ID NO:92, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0047, wherein the first polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:93 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0048, wherein the first polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:94 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0060, wherein the first polypeptide consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:95 is linked to a second a polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0061, wherein the first polypeptide consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:96, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0019, wherein the first polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:86, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, NYF-0014, wherein the first polypeptide consisting of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:149, is linked to a second polypeptide consisting of an amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, and NYF-0082, in which the first polypeptide consists of the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:150 is linked to a second polypeptide consisting of the amino acid sequence of amino acids 21-246 of the amino acid sequence of SEQ ID NO:84, can be given as an example of preferred taFv-Fc type heterodimer bispecific molecules according to the present invention.

Further examples of preferred taFv-Fc type heterodimer bispecific molecules of the present invention include NYZ-0038, wherein a first polypeptide consisting of an amino acid sequence of amino acids 20-745 of the amino acid sequence of SEQ ID NO:155 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84, NYZ-0082, wherein a first polypeptide consisting of an amino acid sequence of amino acids 20-750 of the amino acid sequence of SEQ ID NO:156 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84, and NYZ-0083, wherein a first polypeptide consisting of an amino acid sequence of amino acids 20-750 of the amino acid sequence of SEQ ID NO:157 is linked to a second polypeptide consisting of an amino acid sequence of amino acids 20-246 amino acid sequence SEQ ID NO:84.

Of these, NYF-0023, NYF-0047, NYF-0048, NYF-0060, NYF-0061, NYZ-0038, NYZ-0082 and NYZ-0083 exhibit excellent biological activity, physical properties and the like, and thus are particularly preferable.

In addition, the taFv of the first antibody may be attached to one Fc dimer by a linker or directly associated with it without a linker, and the Fab of the first antibody or the second antibody may be attached to the other Fc by a linker or directly associated with it without a linker. In such a bispecific molecule, the Fab is added to a site upstream of the Fc region (ii) (solid) of the second taFv-Fc-type heterodimer polypeptide. Such a bispecific molecule is called a taFv-Fab-Fc-type heterodimer or taFv-Fab-Fc-type heterodimer bispecific molecule (Fig. 3).

In the present invention, for example, taFv anti-HLA-A2/NY-ESO scFv, taFv anti-CD3 scFv and Fab HLA/NY-ESO can be used to incorporate into a bispecific taFv-Fab-Fc type heterodimer molecule.

The taFv-Fab-Fc type heterodimer bispecific molecule preferably comprises (a) a first polypeptide comprising an scFv that specifically binds to human HLA/NY-ESO, an scFv that specifically binds to CD3, and an Fc region (i) of an immunoglobulin in this order from the N terminus to the C terminus, a second polypeptide consisting of an immunoglobulin heavy chain including the Fc region (ii), and a third polypeptide consisting of an immunoglobulin light chain. More preferably, (b) the second polypeptide is linked to the third polypeptide, and (c) the first polypeptide is linked to the second polypeptide at the Fc region (i) and the Fc region (ii). Fig. 3(e) shows an example of the taFv-Fab-Fc type heterodimer bispecific molecule, Fig. 3(f) shows the first polypeptide, Fig. 3(h) shows the second polypeptide, and Fig. 3(i) shows the third polypeptide. As shown in Fig. 3(e), the Fc region (ii) (shaded) of the second polypeptide, consisting of an immunoglobulin heavy chain, including the Fc region (ii), binds to the Fc region (i) of the first polypeptide, and the immunoglobulin light chain is further attached to the second polypeptide. Such a preferred bispecific TaFv-Fab-Fc type heterodimer molecule may consist of a bispecific taFv-Fc type heterodimer molecule comprising taFv and an immunoglobulin Fc region, and Fab linked thereto. Examples of amino acid sequences included in the first polypeptide of the preferred taFv-Fab-Fc type heterodimer bispecific molecule include the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:85, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:87, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:88, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:89, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:90, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:91, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:92, the amino acid sequence of amino acids 21-511 of the amino acid sequence of SEQ ID NO:93, the amino acid sequence of amino acids 21-511 of the amino acid sequence sequence SEQ ID NO:94, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:95, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:96, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:86, the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:149, and the amino acid sequence of amino acids 21-511 of the amino acid sequence SEQ ID NO:150. The first polypeptide included in the more preferred taFv-Fab-Fc type heterodimer bispecific molecule consists of an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:85, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:87, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:88, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:89, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:90, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:91, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:92, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:93, an amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:94, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:95, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:96, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:86, the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:149, or the amino acid sequence of amino acids 21-745 of the amino acid sequence of SEQ ID NO:150.

The second polypeptide included in the preferred taFv-Fab-Fc type heterodimer bispecific molecule of the present invention comprises a heavy chain variable region of a human or humanized antibody, a CH1 region, a hinge region and a mutant Fc. The second polypeptide included in the more preferred taFv-Fab-Fc type heterodimer bispecific molecule comprises an amino acid sequence of amino acids 20-242 of the amino acid sequence of SEQ ID NO:99.

The third polypeptide included in the preferred taFv-Fab-Fc type heterodimer bispecific molecule of the present invention comprises a variable region and a constant region of a light chain of a human or humanized antibody. The third polypeptide included in the more preferred taFv-Fab-Fc type heterodimer bispecific molecule comprises an amino acid sequence of amino acids 21-131 of the amino acid sequence of SEQ ID NO:100.

Of the second polypeptide included in such a preferred taFv-Fab-Fc type heterodimer bispecific molecule, the variable region, the CH1 region and the third polypeptide constitute a Fab, wherein the Fab is preferably an anti-HLA/NY-ESO antibody, such as Fab NYA-0001.

In the present invention, for example, an anti-HLA-A2/NY-ESO scFv, an anti-CD3 scFv, an HLA/NY-ESO Fab or an anti-CD3 Fab can be included in a bispecific Fc-type scFv-Fab heterodimer molecule.

The bispecific scFv-Fab-Fc type heterodimer molecule preferably comprises (a) a first polypeptide comprising an scFv that specifically binds to human HLA/NY-ESO, a variable region of an antibody heavy chain and a constant region CH1 of an antibody that specifically binds to CD3, and an Fc region (i) of an immunoglobulin in this order from N terminus to C terminus, a second polypeptide comprising an immunoglobulin hinge region and an Fc region (ii), and a third polypeptide consisting of an antibody light chain comprising a variable region and a constant region. More preferably, (b) the first polypeptide is linked to the second polypeptide via the Fc region (i) and the Fc region (ii), and the first polypeptide is linked to (the antibody light chain) the third polypeptide via the variable region of the antibody heavy chain and the constant region CH1 of the first polypeptide. The Fc regions of the first polypeptide and the second polypeptide may be wild-type regions or may comprise a mutation that forms a heterodimer. Fig. 6A(c) shows an example of a bispecific scFv-Fab-Fc-type heterodimer molecule. In Fig. 6A(c), the right half shows the first polypeptide and the third polypeptide, and the left half shows the second polypeptide. As shown in Fig. 6A(c), the Fc region (i) of the first polypeptide is linked to the Fc region (ii) of the second polypeptide (solid), and the first polypeptide is linked to the third polypeptide. In Fig. 6A(c), the scFv indicated by a positive slope is an anti-HLA-A2/NY-ESO scFv, and each Fab indicated by an empty, checkered pattern, or horizontal lines is an anti-CD3 Fab.

An example of an amino acid sequence included in the first polypeptide included in the preferred bispecific scFv-Fab-Fc type heterodimer molecule is the amino acid sequence of amino acids 21-394 of the amino acid sequence of SEQ ID NO:160. A more preferred example thereof is the amino acid sequence of amino acids 20-724 of the amino acid sequence of SEQ ID NO:160.

An example of another amino acid sequence included in the first polypeptide included in the preferred bispecific scFv-Fab-Fc type heterodimer molecule is the amino acid sequence of amino acids 21-389 of the amino acid sequence of SEQ ID NO:197. A more preferred example thereof is the amino acid sequence of amino acids 20-719 of the amino acid sequence of SEQ ID NO:197.

An example of another sequence included in the first polypeptide included in the preferred bispecific scFv-Fab heterodimer molecule of the Fc type is the amino acid sequence of amino acids 21-389 of the amino acid sequence of SEQ ID NO:198. A more preferred example thereof is the amino acid sequence of amino acids 20-719 of the amino acid sequence of SEQ ID NO:198.

The second polypeptide included in the preferred bispecific scFv-Fab-Fc-type heterodimer molecule of the present invention comprises a hinge region and a mutant Fc derived from a human antibody. The second polypeptide included in the more preferred bispecific scFv-Fab-Fc-type heterodimer molecule comprises the amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84.

The third polypeptide included in the preferred bispecific Fc-type scFv-Fab heterodimer molecule of the present invention comprises a light chain derived from a human antibody. The third polypeptide included in the more preferred bispecific Fc-type scFv-Fab heterodimer molecule comprises, for example, the amino acid sequence of amino acids 21-127 of the amino acid sequence of SEQ ID NO: 161. An even more preferred example thereof is the amino acid sequence of amino acids 21-233 of the amino acid sequence of SEQ ID NO: 161.

Of these, NYZ-1010, in which the first polypeptide consists of an amino acid sequence of amino acids 20-724 of the amino acid sequence of SEQ ID NO:160, the second polypeptide consists of an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84, and the third polypeptide consisting of an amino acid sequence of amino acids 21-233 of the amino acid sequence of SEQ ID NO:161, are linked to each other, can be given as an example of a preferred bispecific Fc-type scFv-Fab heterodimer molecule of the present invention.

In addition, NYZ-1007, in which the first polypeptide consists of an amino acid sequence of amino acids 20-719 of the amino acid sequence of SEQ ID NO:197, the second polypeptide consists of an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84, and the third polypeptide consists of an amino acid sequence of amino acids 21-233 of the amino acid sequence of SEQ ID NO:161, are linked to each other, can be given as an example of a preferred bispecific Fc-type scFv-Fab heterodimer molecule of the present invention.

In addition, NYZ-1017, in which the first polypeptide consists of an amino acid sequence of amino acids 20-719 of the amino acid sequence of SEQ ID NO:198, the second polypeptide consists of an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84, and the third polypeptide consists of an amino acid sequence of amino acids 21-233 of the amino acid sequence of SEQ ID NO:161, are linked to each other, can be given as an example of a preferred bispecific Fc-type scFv-Fab heterodimer molecule of the present invention.

At least 1, 2 or 3 peptides included in the bispecific molecule of the present invention may be a "deletion mutant" described above. In particular, 1 or 2 (or more) amino acids at its carboxyl terminus (in particular at the carboxyl terminus derived from the heavy chain of the antibody) may be mutated (also may be deleted). For example, the carboxyl terminus of the amino acid sequence of the first polypeptide included in NYZ-1010, which is a preferred bispecific scFv-Fab-Fc-type heterodimer molecule of the present invention, may be any of Lys at position 724 of SEQ ID NO: 160, Gly at position 723 as a result of amino acid deletion, or a mixture of the first polypeptides comprising Lys and Gly at the carboxyl terminus. Furthermore, the carboxyl terminus of the amino acid sequence of the second polypeptide included in the preferred bispecific Fc-type scFv-Fab heterodimer molecule of the present invention may be any of Lys at position 246 of SEQ ID NO:84, Gly at position 245 as a result of amino acid deletion, or a mixture of second polypeptides including Lys and Gly at the carboxyl terminus.

The scFv and Fab included in the bispecific molecule of the present invention are preferably derived from a humanized antibody or a human antibody, and the Fc is preferably derived from a human antibody.

In the variable region of the bispecific molecule of the present invention, the variable region of the heavy chain and the variable region of the light chain may be attached in this order from the amino terminus of the antibody. Alternatively, the variable region of the light chain and the variable region of the heavy chain may be attached in this order. A linker may optionally be present between the two variable regions. A glycine residue may optionally be present at the amino terminus of the amino terminal variable region. In a tandem bispecific molecule such as an scFv, a linker, a FLAG tag and/or a His tag may optionally be attached to the carboxyl terminus of the variable region of the carboxyl terminus. In a preferred embodiment, for example, the variable region of the heavy chain, the first linker, the variable region of the light chain, the second linker, the FLAG tag and the His tag are attached in this order from the amino terminus.

Examples of linkers include single-chain polypeptide, single-chain oligopeptide, and synthetic products such as PEG, nucleotide, sugar chain, and compounds. Other known linkers can also be used without particular limitations as long as they can attach two polypeptides to each other.

The peptide linker may contain, for example, from 5 to 30 amino acids. When the bispecific molecule contains multiple linkers, all peptide linkers may be of the same length or peptide linkers of different lengths may be used.

An example of a peptide linker is the repeat (Gly⋅Gly⋅Gly⋅Gly⋅Ser) (SEQ ID NO:161). One or more amino acid residues other than Gly and Ser may be added to these.

Among the constructs (formats) of the multispecific antibody and, in particular, the bispecific antibody of the present invention as described above, taFv-Fc type heterodimer, taFv-Fab-Fc type heterodimer and scFv-Fab-Fc type heterodimer are preferable, and taFv-Fc type heterodimer is more preferable. A format in which the anti-HLA/NY-ESO scFv and the anti-CD3 scFv are arranged in such an order from the N terminus to the C terminus (taFv-Fc type heterodimer) is more preferable than a format in which the anti-CD3 scFv and the anti-HLA/NY-ESO scFv are arranged in such an order (taFv (inverted)-Fc type heterodimer) (e.g., Example 11). Another example of a more preferable format is scFv-Fab-Fc type heterodimer.

The present invention also encompasses a molecule that comprises an amino acid sequence encoded by a nucleotide sequence included in a polynucleotide that hybridizes under stringent conditions to the complementary strand of a polynucleotide containing a nucleotide sequence encoding an amino acid sequence included in the molecule of the present invention and that binds to HLA/NY-ESO and further binds to CD3.

The present invention also encompasses a molecule comprising an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to an amino acid sequence included in a molecule of the present invention and which binds to HLA/NY-ESO and also binds to CD3.

The antibody of the present invention, the binding fragment thereof, and the multispecific antibody comprising the same have excellent biological activity, physicochemical properties (hereinafter referred to as "physical properties"), safety, pharmacokinetics, and other properties. (a) Examples of the biological activity or index thereof include antigen-binding activity, in vitro cytotoxicity, and in vivo antitumor activity. For example, the dissociation constant (KD value) for HLA/NY-ESO is 100 nM or less, or 50 nM or less, preferably 20 nM or less, or 10 nM or less, and more preferably 5 nM or less. The _EC50 value of cytotoxicity exhibited by using human peripheral blood mononuclear cells as effector cells against human U266B1 and/or NCI-H1703 cells endogenously expressing NY-ESO is 20 nM or less, preferably 10 nM or less, and more preferably 5 nM or less (the in vitro cytotoxicity can be analyzed and measured according to the method described in Example 8, although the method is not limited thereto). For example, 0.1 ml of a suspension of human squamous cell lung cancer cells NCI-H1703 (6× ¹⁰⁷ cells/ml) is subcutaneously administered to NOG mice, 0.2 ml of a suspension of human peripheral blood mononuclear cells (3.75× ¹⁰⁷ cells/ml) is administered intravenously after 4 days, and antibody administration is started after 14 days and carried out once a week on 3 occasions. Thereafter, the tumor volume is measured. The tumor growth inhibitory activity compared with the control group administered with the solvent is 50% or higher, preferably 75% or higher, and more preferably 90% or higher (the in vivo antitumor activity can be analyzed and measured according to the method described in Example 9, although the method is not limited thereto). (b) Impurities contained in biopharmaceutical products are related to the safety of the drug. Therefore, it is necessary to establish an appropriate standard and control the amount of impurities during production and during storage. In particular, HMWS (aggregate) is a major impurity and is associated with immunogenic risk or decreased efficacy of the drug. Therefore, HMWS should be more strictly controlled. For the purpose of impurity control, stability evaluation (whether or not the amount of impurities has increased) over time during and after production and during production should also be performed. Since the period of action of a pharmaceutical product is determined based on the results of a long-term stability test, a longer period of action can be determined for an antibody that exhibits stability over time. Thus, long-term stability can be used as an indicator for selecting a preferred antibody for a biopharmaceutical product. Examples of physicochemical properties in the present invention include acid stability (for inhibiting the formation of HMWS) and stability in solution (for inhibiting the formation of HMWS). An example of another indicator is a high yield of a recombinant cell culture product obtained by introducing a gene encoding an amino acid sequence of the antibody of the present invention, a binding fragment thereof, or a molecule containing it into a suitable host cell for producing them, such as an Expi293F cell. The preferred antibody of the present invention, its antigen-binding fragment, and a multispecific antibody containing it with such physicochemical properties are preferable for the following reasons. That is, the solution containing the antibody, its binding fragment or multispecific antibody can be exposed to acidic conditions, and thus the processes for obtaining it, such as protein A or ion exchange chromatography and virus inactivation, can be carried out or facilitated. In addition, the production of HMWS can be controlled at low levels even in the form of a solution of the antibody, its binding fragment or multispecific antibody, and thus its production, preparation of a drug, distribution or storage of pharmaceutical products containing it can be carried out and facilitated. In addition, pharmaceutical products containing them can be efficiently produced. As for the acid stability, for example, the content of HMWS determined by heating at pH 3.5 at room temperature for 1 hour and then measuring HMWS by size exclusion chromatography is 5% or less, preferably 2% or less, and more preferably 1% or less (the content of HMWS can be measured and determined for evaluation of the acid stability according to the method described in Example 19-1, although the method is not limited thereto). As for the solution stability, for example, the analyte is dissolved to a concentration of 25 mg/mL in a solution containing 25 mM histidine and 5% sorbitol at pH 6.0, the resulting mixture is stored at 25°C for 6 days, and then the HMWS is measured by size exclusion chromatography. The HMC content determined in this manner is 20% or less, preferably 10% or less (the HMC content can be measured and determined to evaluate the stability of the solution according to the method described in Example 19-2, although the method is not limited thereto). The production efficiency or yield can be measured and determined according to the method described in Examples 20 and 21, although the method is not limited thereto. (c) Examples of safety and its indicators include antigen recognition properties and observation during administration. For example, an antibody of interest that recognizes a plurality of amino acids on the wild-type NY-ESO peptide but does not bind to a homologous peptide containing an amino acid sequence similar to but not identical to the amino acid sequence of the wild-type NY-ESO peptide will result in a low risk of side effects caused by off-target effects. In addition, it can be predicted that such an antibody will exhibit low immunogenicity in an immunogenicity screen based on ISPRI web (EpiVax, Inc) and exhibit a low risk of side effects such as cytokine production. In addition, NYF-0023, NYF-0045, NYF-0047, NYF-0048, NYF-0060, NYF-0061, NYZ-0082 or NYZ-1010 included in the bispecific antibody of the present invention are administered to Balb/c mice. As a result, no significant problems with blood half-life are observed, and no weight loss or other obvious toxicity problems are observed. As a result of a single administration of NYZ-0082 or NYZ-1010 to cynomolgus macaques, no significant problem with the half-life in blood is observed, and no change in the general condition, body weight, food intake amount, body temperature and plasma cytokine level is observed as a result of the administration. (d) Examples of kinetics or indices thereof include the half-life in blood. In the present invention, no significant problem with the half-life in blood is observed as a result of the administration of several bispecific antibodies to Balb/c mice or cynomolgus macaques. The antibody of the present invention, the binding fragment thereof and the molecules having such excellent biological activity, physicochemical properties, safety and kinetics can be preferably integrated into a pharmaceutical composition. Preferred examples of the antibody of the present invention or an antigen-binding fragment thereof having the antigen-binding activity described in (a) and the various properties described in (b) include, but are not limited to, NYA-1143, NYA-1163, and NYA-3061. More preferred examples thereof include, but are not limited to, NYA-2047, NYA-2061, NYA-2143, and NYA-3061. Preferred examples of the multispecific antibody of the present invention having the properties (a) to (d) include, but are not limited to, NYF-0016, NYF-0019, NYF-0022, NYF-0023, NYF-0027, NYF-0035, NYF-0044, NYF-0045, NYF-0047, NYF-0048, NYF-00058, NYF-0060, NYF-0061, NYZ-0038, NYZ-0082, NYZ-0088 and NYZ-1010. More preferred examples thereof include, but are not limited to, NYF-0061, NYZ-0038, NYZ-0082, NYZ-0088, NYZ-1007, NYZ-1010, and NYZ-1017.

In the present invention, the "site" to which the antibody binds; i.e., the "site" that is recognized by the antibody, is a partial peptide or a partial higher-order structure of the antigen to which the antibody binds or which is recognized by the antibody. In the present invention, such a site is also called an epitope or an antibody-binding site. Examples of sites to which the anti-HLA/NY-ESO antibody of the present invention binds or sites that are recognized by it on HLA/NY-ESO include a plurality of amino acids and partial higher-order structures in the HLA/NY-ESO peptide.

"An antibody or a binding fragment thereof that binds to the same site" as the antibody of the present invention, or a binding fragment thereof, are also included in the scope of the present invention. "An antibody that binds to the same site" as the antibody is another antibody that binds to a site on an antigen molecule recognized by such said antibody. If the second antibody binds to a partial peptide or a partial conformation of the antigen molecule to which the first antibody binds, binding of the first and second antibodies to the same site can be assessed. When the first antibody of the present invention has the antigen-binding activity described in (a) above, a second antibody that binds to the same site on HLA/NY-ESO is highly likely to have similar activity, and thus such a second antibody is included in the scope of the present invention. An antibody that binds to HLA/NY-ESO competitively with the first antibody of the present invention having the antigen-binding activity described in (a) is within the scope of the present invention. An antibody that binds to a site on HLA/NY-ESO recognized by the monoclonal antibody of the present invention, the antibody that binds to HLA/NY-ESO competitively with the monoclonal antibody of the present invention, and binding fragments thereof preferably have at least one of the in vitro cytotoxicity and in vivo antitumor activity described in (a) and the properties described in (b) to (d), more preferably 3 or more of them, and optimally all of them.

The binding site of the antibody can be determined by a method well known to a person skilled in the art, such as an immunoassay method. For example, a series of peptides are obtained by removing the amino acid sequence of the antigen from the C or N terminus, respectively, the reactivity of the antibody to it is examined, the recognition site is approximately determined, a shorter peptide sequence is synthesized and the reactivity of the antibody to it is determined. In this way, the binding site can be determined. Alternatively, a specific site or region of the amino acid sequence of the antigen or peptide fragment of the antigen is deleted or replaced by another amino acid sequence, or a mutation is introduced into such a site or region. By examining the reactivity of the antibody to such a peptide, the binding site can be determined. The peptide fragment of the antigen can be obtained, for example, by genetic recombination or peptide synthesis.

When an antibody binds to or recognizes a partial structure of a higher order antigen, such an antigen-binding site of the antibody can be determined by identifying an amino acid residue on the antigen adjacent to the antibody using X-ray crystallography. For example, an antibody or a fragment thereof and an antigen or a fragment thereof are bound to each other, crystallized and subjected to structural analysis. In this way, an amino acid residue on the antigen located at an interaction distance with the antibody can be identified. The interaction distance is 8 Å or less, preferably 6 Å or less, and more preferably 4 Å or less. One or more amino acid residues located at such an interaction distance with the antibody can constitute an antigen-binding site of the antibody (epitope). When there are two or more such amino acid residues, such amino acid residues may not be adjacent to each other in a one-dimensional sequence.

The anti-HLA/NY-ESO antibody of the present invention or a binding fragment thereof specifically recognizes a plurality of amino acids in the amino acid sequence of HLA/NY-ESO. An antibody or a binding fragment thereof that recognizes a plurality of amino acids, an antibody or a binding fragment thereof that binds to HLA/NY-ESO competitively with the antibody of the present invention or a binding fragment thereof, and an antibody or a binding fragment thereof that has an interaction distance with such a plurality of amino acids are within the scope of the present invention. A multispecific antibody comprising such an antibody or a binding fragment thereof is also within the scope of the present invention.

6. Pharmaceutical composition

The present invention includes an anti-cancer agent comprising, as an active ingredient, the anti-HLA/NY-ESO antibody of the present invention or a multispecific molecule that binds to HLA/NY-ESO.

The anticancer agent of the present invention can be used to treat one or two or more types of cancer selected from carcinoma, sarcoma, lymphoma, leukemia, myeloma, germinoma, brain tumor, carcinoid, neuroblastoma, retinoblastoma and nephroblastoma. Specific examples of carcinoma include renal cell cancer, melanoma, squamous cell carcinoma, basal cell carcinoma, conjunctival cancer, oral cavity cancer, laryngeal cancer, pharyngeal cancer, thyroid cancer, lung cancer (non-small cell lung cancer (adenocarcinoma, epidermoid carcinoma, large cell carcinoma, and small cell lung cancer), breast cancer, esophageal cancer, stomach cancer, duodenal cancer, small bowel cancer, colon cancer, rectal cancer, appendiceal cancer, anal cancer, liver cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, adrenal cancer, bladder cancer, prostate cancer, uterine cancer, and vaginal cancer. Specific examples of sarcoma include liposarcoma, angiosarcoma, chondrosarcoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, undifferentiated pleomorphic sarcoma, myxofibrosarcoma, malignant peripheral neurilemoma, retroperitoneal sarcoma, synoviosarcoma, uterine sarcoma, gastrointestinal stromal tumor, leiomyosarcoma, and epithelioid sarcoma. Specific examples of lymphoma include B-cell lymphoma, T/NK cell lymphoma, and Hodgkin lymphoma. Specific examples of leukemia include myelogenous leukemia, lymphocytic leukemia, myeloproliferative disorder, and myelodysplastic syndrome. Specific examples of myeloma include multiple myeloma. Specific examples of germinoma include testicular cancer and ovarian cancer. Specific examples of brain tumor include neuroglioma and meningioma.

The anti-cancer agent of the present invention may contain an anti-HLA/NY-ESO antibody or a multispecific molecule that binds to HLA/NY-ESO in an amount effective for treatment, as well as pharmaceutically acceptable carriers, diluents, solubilizers, emulsifiers, preservatives, additives and the like. "Pharmaceutically acceptable carriers" and the like can be appropriately selected from a wide range according to the type of the target disease and the dosage form of the drug. The administration route of the anti-cancer agent of the present invention can be appropriately selected. For example, the anti-cancer agent can be administered by injection, and local injection, intraperitoneal injection, selective intravenous infusion, intravenous injection, subcutaneous injection, organ perfusate and the like can be used. In addition, an injection solution can be formulated using a carrier including a salt solution, a glucose solution or a mixture of salt water and a glucose solution, various types of buffer solutions and the like. Alternatively, the powder can be prepared and mixed with a liquid carrier to prepare an injection solution before use.

Other routes of administration can be appropriately selected along with the formulation. For example, oral solutions, powders, pills, capsules, tablets and the like can be used for oral administration. As for oral solutions, liquid oral preparations such as suspensions and syrups can be prepared using water, saccharides such as sucrose, sorbitol and fructose, glycols such as PEG, oils such as sesame oil and soybean oil, preservatives such as alkyl parahydroxybenzoates, flavors such as strawberry and peppermint flavors and the like. Powders, pills, capsules and tablets can be prepared using excipients such as lactose, glucose, sucrose and mannitol, disintegrating agents such as starch and sodium alginate, lubricating agents such as magnesium stearate and talc, binders such as polyvinyl alcohol, hydroxypropyl cellulose and gelatin, surfactants such as fatty acid esters, plasticizers such as glycerol and the like. Tablets and capsules are preferred unit dosage forms for the composition of the present invention, since they are easy to administer. Solid manufacturing carriers are used for the production of tablets and capsules.

The effective dose of anti-HLA/NY-ESO antibody or multispecific molecule that binds to HLA/NY-ESO used for treatment can be changed according to the characteristics of the symptoms to be treated and the age and condition of the patient, and can be finally changed by the attending physician. For example, a single dose is 0.0001 mg to 100 mg per kg of body weight. The given dose can be administered once every 1 to 180 days, or the dose can be divided into two doses, three doses, four doses or more doses per day at appropriate intervals.

The present invention also encompasses a polynucleotide comprising an amino acid sequence included in the anti-HLA/NY-ESO antibody of the present invention or a multispecific molecule binding to HLA/NY-ESO, a vector comprising such a polynucleotide, a cell comprising such a polynucleotide or a vector, and a pharmaceutical composition comprising any of the polynucleotide, the vector, and the cell as an active ingredient. Such a pharmaceutical composition is preferably an anticancer agent.

In addition, the pharmaceutical composition of the present invention can be used in combination with other therapeutic agents or therapeutic methods. Examples of other therapeutic agents or therapeutic methods include, but are not limited to, chemotherapeutic agents, radiation therapy, and biopharmaceutical products. The pharmaceutical composition of the present invention and other therapeutic agents can be administered simultaneously or in different regimens as a single preparation or two or more different preparations containing the pharmaceutical composition of the present invention. The administration of the pharmaceutical composition can be carried out in combination with other therapeutic methods.

Examples

The present invention will now be described in more detail with reference to examples, although the present invention is not limited to these examples.

In the following examples, the genetic engineering procedures are carried out according to the method described in Molecular Cloning (Sambrook, J., Fritsch, E. F. and Maniatis, T., Cold Spring Harbor Laboratory Press, 1989) and the methods described in other experimental protocols used by those skilled in the art, unless otherwise indicated. When a commercially available reagent or kit is to be used, the procedures are followed in accordance with the instructions of such commercial product. The synthesis of primers required for gene synthesis or vector construction is outsourced, as necessary (Fasmac Co., Ltd., Thermo Fisher Scientific and Eurofins Genomics).

(Example 1) Preparation of anti-HLA/NY-ESO antibody derived from human antibody phage library

1)-1 Obtaining HLA/NY-ESO antigen protein

Inclusion bodies obtained from E. coli cells (BL21 (DE3), Agilent Technologies) expressing HLA-A ^* 0201 (GenBank: ASA47534.1) were truncated (adding biotin ligase recognition sequence: SEQ ID NO:33), and β2-microglobulin (UniProtKB-P61769: SEQ ID NO:34) and NY-ESO peptide: SLLMWITQC (SEQ ID NO:1 from the sequence listing) were refolded at a high dilution factor, and then the HLA-A ^* 0201/β2-microglobulin/NY-ESO peptide complex (hereinafter referred to as “HLA/NY-ESO”) was prepared using gel filtration columns (Superdex 200 10/300, GE Healthcare).

As a negative control antigen, the HLA-A ^* 0201/β2-microglobin/MAGEC-1 peptide complex (hereinafter referred to as “HLA/MC1”) was prepared by refolding using the MAGEC-1 peptide: ILFGISLREV (SEQ ID NO:2 of the Sequence Listing). Next, the obtained HLA/NY-ESO and HLA/MC1 were biotinylated with E. coli biotin ligase, and then fractionated using gel filtration columns (Superdex 200 10/300, GE Healthcare) to obtain biotinylated proteins.

1)-2 Isolation of scFv capable of binding to HLA/NY-ESO

HLA/NY-ESO-binding scFv was isolated from a human antibody phage library. Phages were first added to biotinylated HLA/MC1-immobilized Dynabeads Streptavidin M-280 (Thermo Fisher Scientific) and unbound phages were collected. Phages were then added to biotinylated HLA/NY-ESO-immobilized Dynabeads Streptavidin M-280Ag and unbound phages were removed by washing using a magnetic stand (DynaMag-2, Thermo Fisher Scientific).

Subsequently, E. coli (XL-1 Blue, Agilent Technologies) were infected with HLA/NY-ESO-linked phages, and the phages were collected and amplified. After a total of 3 pannings, the phages were transferred from the polyclonal phagemid into an E. coli expression vector containing a FLAG tag and a His tag added to the carboxyl terminus of the scFv, E. coli was transformed, the scFv was expressed in the presence of IPTG (Sigma-Aldrich), and the expressed scFv was screened by ELISA.

1)-3 Screening of HLA/NY-ESO-related scFv by ELISA

NeutrAvidin (Life Technologies) diluted to 1 μg/ml in PBS (0.01 M phosphate-buffered saline containing 0.138 M sodium chloride and 0.0027 M potassium chloride (pH 7.4), Sigma-Aldrich) was added at 50 μl each to a 384-well Maxi-sorp plate (Black, Nunc), and the plate was incubated at 4°C overnight for immobilization. The plate was washed 3 times with PBS (ELISA buffer) containing 0.05% Tween-20 (BioRad), biotinylated HLA/NY-ESO diluted to 1 μg/ml in PBS and used in Example 1)-2 was added, and the resulting mixture was stirred at room temperature for 1 h. The plate was washed 3 times with ELISA buffer, blocked with Blocker Casein (Thermo Fisher Scientific), and then washed 3 times with ELISA buffer. After that, the culture solution of scFv-expressing E. coli was added, and the reaction was allowed to proceed at room temperature for 2 hours. After the resulting mixture was washed 3 times with ELISA buffer, 50 μl of horseradish peroxidase (HRP)-labeled anti-FLAG antibody (Sigma-Aldrich) diluted 5000-fold with ELISA buffer was added, and the reaction was allowed to proceed at room temperature for 1 hour. After the resulting product was washed 5 times with ELISA buffer, SuperSignal Pico ELISA chemiluminescent substrate (Thermo Fisher Scientific) was added, chemiluminescence was analyzed after 10 minutes using a plate reader (Envision 2104 Multilabel Reader, Perkin Elmer), and positive clones were selected in an HLA/NY-ESO-binding ELISA.

1)-4 Determination of the nucleotide sequence and amino acid sequence of the ELISA-positive clone NY-R119

Among the ELISA-positive clones obtained in 1)-3, NY-R119 was selected as an scFv exhibiting high binding affinity to HLA/NY-ESO and excellent recognition specificity. The nucleotide sequences of the variable regions of the heavy chain and light chain of NY-R119 were analyzed by the dye terminator method (BigDye®, Terminator v3.1, Thermo Fisher Scientific). The sequences of the primers used for the sequence analysis are shown below.

Primer A: 5'-CTCTTCGTATTACGCCAGCTGGCGA-3' (SEQ ID NO:3 from the sequence listing (Fig. 10))

Primer B: 5'-ATAACAATTTCACACAGGAAACAGCTATGA-3' (SEQ ID NO:4 from the sequence listing (Fig. 11))

SEQ ID NO:5 (Fig. 12) shows the determined nucleotide sequence of the cDNA encoding the variable region of the heavy chain of NY-R119, and SEQ ID NO:6 (Fig. 13) shows the determined amino acid sequence thereof.

SEQ ID NO:7 (Fig. 14) shows the determined nucleotide sequence of the cDNA encoding the light chain variable region of NY-R119, and SEQ ID NO:8 (Fig. 15) shows the determined amino acid sequence thereof.

The CDR sequences of NY-R119 according to the CDR definition provided by IMGT are shown as follows: CDRH1: SEQ ID NO:54 (Fig. 61); CDRH2: SEQ ID NO:55 (Fig. 61); CDRH3: SEQ ID NO:56 (Fig. 61); CDRL1: SEQ ID NO:57 (Fig. 61); CDRL2: SEQ ID NO:58 (Fig. 61); and CDRL3: SEQ ID NO:59 (Fig. 61).

1)-5 Receiving NYA-0001

1)-5-1 Construction of expression vector NYA-0001

To prepare various samples for evaluation, a mammalian cell expression vector of NY-R119 was constructed. NY-R119 expressed in cultured mammalian cells was designated as “NYA-0001”. The amino acid sequences constituting the scFv region, the variable regions of the heavy chain and light chain, and CDRH1-3 and CDRL1-3 of NY-R119 were identical to those of NYA-0001. The DNA fragment encoding NYA-0001 was inserted into a mammalian cell expression vector containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone using the In-Fusion HD Cloning Kit (CLONTECH) to construct the expression vector of NYA-0001.

The nucleotide sequence of the constructed NYA-0001 expression vector was reanalyzed, and the nucleotide sequence of the full-length NYA-0001 was found to be the nucleotide sequence shown in SEQ ID NO: 69 of the sequence listing (Fig. 70). Furthermore, based on the nucleotide sequence, the amino acid sequence of the entire NYA-0001 encoded in this manner was found to be the amino acid sequence shown in SEQ ID NO: 70 (Fig. 71). In such an amino acid sequence, amino acids 1 to 19 constitute a signal sequence, amino acids 21 to 266 constitute NYA-0001, and amino acids 267 to 292 constitute a Flag-His tag. In addition, amino acids 46 to 53 constitute CDRH1, amino acids 71 to 78 constitute CDRH2, and amino acids 117 to 129 constitute CDRH3. Additionally, amino acids 181-188 make up CDRL1, amino acids 206-208 make up CDRL2, and amino acids 245-256 make up CDRL3.

1)-5-2 Expression and purification of NYA-0001

Expi293F cells (Thermo Fisher Scientific) were subcultured according to the instructions. A culture solution of logarithmic Expi293F cells was diluted to 2.5 x 10 ⁶ cells/mL in Expi293 Expression Medium (Thermo Fisher Scientific) and used to produce NYA-0001. NYA-0001 expression vector (0.3 mg) and 0.9 mg polyethyleneimine (Polyscience #24765) were added to 20 mL of Opti-Pro SFM medium (Thermo Fisher Scientific). The mixture was gently mixed, left for five minutes, and then added to Expi293F cells. The culture supernatant, obtained by shaking the culture in an incubator at 37°C in the presence of 8% CO ₂ at 135 rpm for 6 days, was filtered through a 0.2 μm filter (Millipore). In this way, the culture supernatant of NYA-0001 was obtained. Purification was carried out by elution and concentration using Ni Sepharose excel (GE Healthcare), and then filtration through a gel filtration column (Superdex 200 Increase, GE Healthcare) equilibrated with 25 mM histidine, 300 mM NaCl and 5% sorbitol, pH 6.0. The purified protein samples were subjected to analytical size exclusion chromatography (SEC), the degree of purification and concentration were determined, and then the samples were subjected to various types of evaluation.

(Example 2) Obtaining the mutant NYA-0001

2)-1 Obtaining mutant NYA-0001

A phage library is constructed by a method in which the NYA-0001 gene is used as a template and a mutation is introduced by PCR (i.e., error-based libraries) or by a method in which an oligomer is synthesized by random mutation of 20 types of amino acids for each of all CDR residues (i.e., oligonucleotide-based libraries), clones with high binding affinity are selected, NYA-0060, NYA-0068, and NYA-0082 are obtained as high binding mutants, and the affinity and nucleotide sequences thereof are then determined.

SEQ ID NO:9 (Fig. 16) shows the nucleotide sequence of the cDNA encoding the variable region of the heavy chain of NYA-0060, and SEQ ID NO:10 (Fig. 17) shows the amino acid sequence thereof.

SEQ ID NO:11 (Fig. 18) shows the nucleotide sequence of the cDNA encoding the light chain variable region of NYA-0060, and SEQ ID NO:12 (Fig. 19) shows the amino acid sequence thereof.

SEQ ID NO:13 (Fig. 20) shows the nucleotide sequence of the cDNA encoding the variable region of the heavy chain of NYA-0068, and SEQ ID NO:14 (Fig. 21) shows the amino acid sequence thereof.

SEQ ID NO:15 (Fig. 22) shows the nucleotide sequence of the cDNA encoding the light chain variable region of NYA-0068, and SEQ ID NO:16 (Fig. 23) shows the amino acid sequence thereof.

SEQ ID NO:17 (Fig. 24) shows the nucleotide sequence of the cDNA encoding the variable region of the heavy chain of NYA-0082, and SEQ ID NO:18 (Fig. 25) shows the amino acid sequence thereof.

SEQ ID NO:19 (Fig. 26) shows the nucleotide sequence of the cDNA encoding the light chain variable region of NYA-0082, and SEQ ID NO:20 (Fig. 27) shows the amino acid sequence thereof.

2)-2 Generation of high-affinity binding mutants using mutation sites and combinations identified in NYA-0060, NYA-0068, and NYA-0082.

DNA fragments encoding NYA-1163 and NYA-2023 with the NYA-0060 and NYA-0068 mutation sites were inserted into a mammalian cell expression vector containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone using the In-Fusion HD Cloning Kit (CLONTECH) to construct scFv expression vectors for mammalian cell culture.

According to the mutation sites and their combinations identified in NYA-0060, NYA-0068 and NYA-0082, NYA-2027, NYA-1143 and NYA-2143 were constructed. The DNA fragments encoding the target scFv were inserted into the expression vector NYA-0001 constructed in 1)-5 by site-directed mutagenesis or a mammalian cell expression vector containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone using the In-Fusion HD Cloning Kit (CLONTECH) for construction of scFv expression vectors for mammalian cell culture.

The nucleotide sequences of the constructed scFv expression vectors were reanalyzed, and it was found that the nucleotide sequence of full-length NYA-1163 was the nucleotide sequence shown in SEQ ID NO:21 of the sequence listing (Fig. 28).

The nucleotide sequence of full-length NYA-2023 was found to be the nucleotide sequence shown in SEQ ID NO:22 of the sequence listing (Fig. 29).

The nucleotide sequence of full-length NYA-2027 was found to be the nucleotide sequence shown in SEQ ID NO:23 of the sequence listing (Fig. 30).

The nucleotide sequence of full-length NYA-1143 was found to be the nucleotide sequence shown in SEQ ID NO:24 of the sequence listing (Fig. 31).

The nucleotide sequence of full-length NYA-2143 was found to be the nucleotide sequence shown in SEQ ID NO:25 of the sequence listing (Fig. 32).

In addition, based on the above nucleotide sequences, the amino acid sequences of the full-length NYA-1163, NYA-2023, NYA-2027, NYA-1143 and NYA-2143 encoded by them are identified.

The amino acid sequence of full-length NYA-1163 is the amino acid sequence shown in SEQ ID NO:26 of the sequence listing (Fig. 33). In such amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-1163, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of the full-length NYA-2023 is the amino acid sequence shown in SEQ ID NO:27 of the sequence listing (Fig. 34). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2023, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of the full-length NYA-2027 is the amino acid sequence shown in SEQ ID NO:28 of the sequence listing (Fig. 35). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2027, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-1143 is the amino acid sequence shown in SEQ ID NO:26 of the sequence listing (Fig. 36). In this amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-1143, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-2143 is the amino acid sequence shown in SEQ ID NO:30 of the sequence listing (Fig. 37). In this amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2143, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

All CDR sequences of NYA-1163 are identical to the CDR sequences of NYA-0001. The CDR sequences of NYA-1143, NYA-2143, and NYA-2023 are identical to each other, and these sequences are represented as follows: CDRH1: SEQ ID NO:54 (Fig. 61); CDRH2: SEQ ID NO:55 (Fig. 61); CDRH3: SEQ ID NO:56 (Fig. 61); CDRL1: SEQ ID NO:60 (Fig. 62); CDRL2: SEQ ID NO:58 (Fig. 61); and CDRL3: SEQ ID NO:59 (Fig. 61). The CDR sequences of NYA-2027 are represented as follows: CDRH1: SEQ ID NO:54 (Fig. 61); CDRH2: SEQ ID NO:55 (Fig. 61); CDRH3: SEQ ID NO:56 (FIG. 61); CDRL1: SEQ ID NO:57 (FIG. 61); CDRL2: SEQ ID NO:58 (FIG. 61); and CDRL3: SEQ ID NO:61 (FIG. 63).

In addition, NYA-1154 containing the binding sites and mutations observed during the screening of high affinity binding clones in combination was constructed and introduced into the expression vector NYA-0001 by site-directed mutagenesis to construct an expression vector NYA-1154 containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone. The nucleotide sequences of the constructed scFv expression vectors were reanalyzed and found to be the nucleotide sequence shown in SEQ ID NO: 31 (Fig. 38). Based on the above nucleotide sequences, the amino acid sequence of the full-length NYA-1154 encoded by them was identified (SEQ ID NO: 32 (Fig. 39)). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-266 constitute NYA-1154, and amino acids 267-292 constitute the Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The CDR sequences of NYA-1154 are shown as follows: CDRH1: SEQ ID NO:54 (Fig. 61); CDRH2: SEQ ID NO:55 (Fig. 61); CDRH3: SEQ ID NO:62 (Fig. 64); CDRL1: SEQ ID NO:57 (Fig. 61); CDRL2: SEQ ID NO:58 (Fig. 61); and CDRL3: SEQ ID NO:63 (Fig. 64).

NYA-1163, NYA-2023, NYA-2027, NYA-1143, NYA-2143, and NYA-1154 were expressed in the same manner as in 1)-5-2 to purify the target scFv constructs. Purified protein samples were subjected to analytical SEC, the degree of purification and concentration were determined, and the samples were subjected to various assays.

2)-3 Obtaining mutant NYA-1143

2)-3-1 Receiving NYA-2035

NYA-2035 was constructed as a mutant of NYA-1143 and introduced into an NYA-1143 expression vector for mammalian cell culture by site-directed mutagenesis to generate an NYA-2035 expression vector. The nucleotide sequence of the constructed scFv expression vector was reanalyzed and found to be the nucleotide sequence shown in SEQ ID NO: 35 (Fig. 42). Based on the above nucleotide sequences, the nucleotide sequence of the full-length NYA-2025 encoded by them was identified (SEQ ID NO: 36 (Fig. 43)). In this amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2035, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3. The CDR sequences of NYA-2035 according to the CDR definition provided by IMGT are shown as follows: CDRH1: SEQ ID NO:54 (Fig. 61); CDRH2: SEQ ID NO:55 (Fig. 61); CDRH3: SEQ ID NO:56 (Fig. 61); CDRL1: SEQ ID NO:64 (Fig. 65); CDRL2: SEQ ID NO:58 (Fig. 61); and CDRL3: SEQ ID NO:59 (Fig. 61).

2)-3-2 Obtaining a CDR NYA-1143 CDR transplant

To improve the physical properties, a CDR mutant of NYA-1143 was constructed. The VH and VL framework sequences of NYA-1143 were compared with the framework sequences of the human subgroup consensus sequence or the germline sequence defined in KABAT et al. (Sequences of Proteins of Immunological Interest, 5th Ed., Public Health Service National Institutes of Health, Bethesda, MD., 1991). As a result, the consensus sequence between the human germline sequence IGHV3_30*15 and the human γ chain subgroup 3 was selected for VH, and the human germline sequence IGLV1-44*01 was selected for VL as acceptors having high identity in the framework regions. Then, the amino acid residues in the framework region of the acceptor were aligned with the amino acid residues of NYA-1143 to identify different amino acid residues.

The three-dimensional model of NYA-1143 was then used to select framework residues for transfer to the acceptor according to the criteria provided by Queen et al. (Proc. Natl. Acad. Sci. U.S.A., 86, 10029-10033, 1989). According to the procedure described above, the CDRs were grafted with mutant amino acid sequences of the VH regions of NYA-1143; i.e., NYA-1143-VH01, NYA-1143-VH02, and NYA-1143-VH03 were constructed. Such amino acid sequences are shown in SEQ ID NO:37 (Fig. 44), SEQ ID NO:38 (Fig. 45), and SEQ ID NO:39 (Fig. 46). As a mutant amino acid sequence of the CDR graft of the VL region of NYA-1143, NYA-1143-VL01 was constructed. Such an amino acid sequence is shown in SEQ ID NO:40 (Fig. 47).

Various scFv constructs were constructed using a combination of VH and VL sequences. An scFv resulting from the replacement of the VH region of NYA-1143 with the amino acid sequence NYA-1143-VH01 was designated NYA-2044. An scFv resulting from the replacement of the VL region of NYA-2044 with the amino acid sequence NYA-1143-VL01 was designated NYA-2045.

The scFv resulting from the replacement of the VH region of NYA-1143 with the amino acid sequence NYA-1143-VH02 is designated as NYA-2047. The scFv resulting from the replacement of the VL region of NYA-2047 with the amino acid sequence NYA-1143-VL01 is designated as NYA-2048.

The scFv resulting from the replacement of the VH region of NYA-1143 with the amino acid sequence NYA-1143-VH03 is designated as NYA-2060. The scFv resulting from the replacement of the VL region of NYA-2060 with the amino acid sequence NYA-1143-VL01 is designated as NYA-2061.

DNA fragments of different engineered NYA-1143 CDR graft mutants were completely synthesized (Fasmac Co., Ltd.) and linked together using the In-Fusion HD Cloning Kit (CLONTECH) to construct an scFv expression vector for mammalian cells containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone.

The nucleotide sequences of the constructed scFv expression vectors were reanalyzed, and it was found that the nucleotide sequence of full-length NYA-2044 is the nucleotide sequence shown in SEQ ID NO:41 of the sequence listing (Fig. 48).

The nucleotide sequence of full-length NYA-2045 was found to be the nucleotide sequence shown in SEQ ID NO:42 of the sequence listing (Fig. 49).

The nucleotide sequence of full-length NYA-2047 was found to be the nucleotide sequence shown in SEQ ID NO:43 of the sequence listing (Fig. 50).

The nucleotide sequence of full-length NYA-2048 was found to be the nucleotide sequence shown in SEQ ID NO:44 of the sequence listing (Fig. 51).

The nucleotide sequence of full-length NYA-2060 was found to be the nucleotide sequence shown in SEQ ID NO:45 of the sequence listing (Fig. 52).

The nucleotide sequence of full-length NYA-2061 was found to be the nucleotide sequence shown in SEQ ID NO:46 of the sequence listing (Fig. 53).

Based on the above nucleotide sequences, the amino acid sequences of the full-length NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060 and NYA-2061 encoded by them were identified. The CDR sequences of NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060 and NYA-2061 were identical to those of NYA-1143.

The amino acid sequence of full-length NYA-2044 is the amino acid sequence shown in SEQ ID NO:47 of the sequence listing (Fig. 54). In such amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2044, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-2045 is the amino acid sequence shown in SEQ ID NO:48 of the sequence listing (Fig. 55). In such amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2045, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-2047 is the amino acid sequence shown in SEQ ID NO: 50 of the sequence listing (Fig. 57). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2047, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-2048 is the amino acid sequence shown in SEQ ID NO:51 of the sequence listing (Fig. 58). In such amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2048, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-2060 is the amino acid sequence shown in SEQ ID NO:52 of the sequence listing (Fig. 59). In such amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2060, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

The amino acid sequence of full-length NYA-2061 is the amino acid sequence shown in SEQ ID NO:53 of the sequence listing (Fig. 60). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-266 constitute NYA-2061, and amino acids 267-292 constitute a Flag-His tag. In addition, amino acids 46-53 constitute CDRH1, amino acids 71-78 constitute CDRH2, and amino acids 117-129 constitute CDRH3. In addition, amino acids 181-188 constitute CDRL1, amino acids 206-208 constitute CDRL2, and amino acids 245-256 constitute CDRL3.

2)-3-3 Expression and purification of NYA-1143 mutants

NYA-2035, NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060, and NYA-2061 obtained in 2)-3-1 and 2)-3-2 were expressed in the same manner as in 1)-5-2 to purify the target scFv constructs. Purified protein samples were subjected to analytical SEC, the degree of purification and concentration were determined, and then the samples were subjected to various assays.

(Example 3) Preparation of scFv reference anti-HLA/NY-ESO antibody

Constructs of scFv antibodies 3M4E5 and T1 with high binding affinity to HLA/NY-ESO (WO2010/106431) are constructed; scFv 3M4E5 is designated as NYC-0003, and scFv T1 is designated as NYC-0004.

The NYC-0003 and NYC-0004 DNA fragments were completely synthesized (Thermo Fisher Scientific), and the scFv expression vector for mammalian cells containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone was constructed using the In-Fusion HD Cloning Kit (CLONTECH).

The nucleotide sequences of the constructed scFv expression vectors were reanalyzed, and the nucleotide sequences of the full-length NYC-0003 and NYC-0004 were found to be the nucleotide sequences shown in SEQ ID NO:65 of the sequence listing (Fig. 66) and its SEQ ID NO:66 (Fig. 67). Based on the nucleotide sequences, the amino acid sequence of the full-length NYC-0003 encoded by them was found to be the amino acid sequence shown in SEQ ID NO:67 (Fig. 68), and the amino acid sequence of the full-length NYC-0004 was found to be the amino acid sequence shown in SEQ ID NO:68 (Fig. 69).

NYC-0003 and NYC-0004 were expressed in the same manner as in 1)-5-2 to purify the target scFv constructs. Purified protein samples were subjected to analytical SEC, the degree of purification and concentration were determined, and then the samples were subjected to various assays.

(Example 4) Evaluation of HLA/NY-ESO binding affinity using Biacore

Using Biacore T200, anti-HLA/NY-ESO scFv was captured as a ligand by immobilized anti-His antibody, and the antigen was analyzed as an analyte. HLA/NY-ESO prepared in 1)-1 was used as an antigen. Anti-His antibody (His Capture kit, GE Healthcare) was immobilized on the CM5 sensor chip (GE Healthcare) according to the kit instructions. Anti-HLA/NY-ESO scFv constructs to be tested diluted to 0.5 μg/mL in HBS-EP+ (GE Healthcare) were contacted with it at 10 μL/min for 60 seconds for immobilization. Thereafter, the samples were added to HLA/NY-ESO analytes diluted to different levels with HBS-EP+ at a flow rate of 30 μl/min for 120 seconds and dissociation was analyzed for 600 seconds. The calculation results obtained by this single-cycle kinetics analysis, K _D , are shown in Table 1. It was found that, compared with the parent antibody NYA-0001, all its mutants exhibited higher binding affinity to HLA/NY-ESO. In addition, compared with NYC-0004, NYA-1143, NYA-2023, NYA-2143, NYA-2044, NYA-2045, NYA-2060, and NYA-2061 were found to exhibit higher binding affinity, and in particular, NYA-1143, NYA-2044, NYA-2045, and NYA-2143 exhibited a K _D of 1 nM or less.

[Table 1]

Clone name K _D (nM) NYA-0001 33.8 NYA-1143 1.0 NYA-1163 13.0 NYA-2023 2.4 NYA-2027 11.2 NYA-2035 3.2 NYA-2044 0.8 NYA-2045 0.8 NYA-2047 5.3 NYA-2048 6.3 NYA-2060 1,1 NYA-2061 1.5 NYA-2143 0.9 NYC-0003 8.1 NYC-0004 2.8

(Example 5) Analysis of the recognized amino acid in the NY-ESO anti-HLA/NY-ESO scFv peptide

The concentration of fused human T2 lymphoblast cells (ATCC) was adjusted to an adequate level in AIM-V medium (Thermo Fisher Scientific) containing 20% FBS, NY-ESO peptide solution (SEQ ID NO:1), NY-ESO point mutant peptides 1F, 2M, 3A, 4A, 5A, 6L, 7F, 8A and 9A (SEQ ID NO:121 (Fig. 122), SEQ ID NO:122 (Fig. 123), SEQ ID NO:123 (Fig. 124), SEQ ID NO:124 (Fig. 125), SEQ ID NO:125 (Fig. 126), SEQ ID NO:126 (Fig. 127), SEQ ID NO:127 (Fig. 128), SEQ ID NO:128 (Fig. 129) and SEQ ID NO:129 (Fig. 130)), or gp100 peptide (SEQ ID NO:130 (Fig. 131)) (Sigma Genosys) dissolved to 5 mM in DMSO was added to a final concentration of 50 μM, or DMSO was added at a rate of 1/100, and the resulting mixture was incubated at 37°C for 4 hours. The resulting mixture was washed twice in AIM-V medium containing 20% FBS, its concentration was adjusted to an adequate level with PBS containing 5% FBS, the LIVE/DEAD Fixable Dead Cell Stain Kit (Thermo Fisher Scientific) was added, and the resulting product was incubated at 4°C for 30 minutes. The cells were washed twice in PBS containing 5% FBS, the washed cells were divided into two groups, the cells in PBS containing 5% FBS were seeded onto a 96-well U-bottom microplate at ¹⁰⁵ cells/well, and the plate was centrifuged followed by removal of the supernatant. The scFv anti-HLA/NY-ESO diluted to 100 nM in PBS containing 5% FBS was added at 25 μl/well to one of the divided cells, and the resulting mixture was incubated at 4°C for 30 minutes. The cells were washed twice in PBS containing 5% FBS, Penta-His Alexa Fluor488 (QIAGEN) diluted in PBS containing 5% FBS was added at 25 μl/well, and the mixture was incubated at 4°C for 30 min. The cells were washed twice in PBS containing 5% FBS, Alexa Fluor488 anti-mouse IgG (Thermo Fisher Scientific) diluted in PBS containing 5% FBS was added at 25 μl/well, and the mixture was incubated at 4°C for 30 min. The cells were washed twice in PBS containing 5% FBS, immobilized with Mildform 10 N (FUJIFILM Wako Pure Chemical Corporation) overnight, and resuspended in PBS containing 5% FBS. To standardize the amount of HLA/peptide complex, HLA-A2 antibody BB7.2-Alexa Fluor 488 diluted to 10 μg/ml in PBS containing 5% FBS or mouse IgG2b-Alexa Fluor 488 at 25 μl/well was added to another group of cells, and the resulting mixture was incubated at 4°C for 30 minutes. The cells were washed 2 times in PBS containing 5% FBS, immobilized with Mildform 10 N (FUJIFILM Wako Pure Chemical Corporation) overnight, and resuspended in PBS containing 5% FBS. The cell suspensions were detected using a flow cytometer (CantoII, Becton Dickinson). Data analysis was performed using Flowjo (Treestar) and the geometric mean fluorescence intensity (gMFI) of Alexa Fluor 488 was measured in the dead cell-removed fraction of T2 cells. The standardized gMFI, indicating the binding affinity of each scFv standardized to the amount of HLA/peptide complex on T2 cells, was determined according to the following equation.

Standardized gMFI=(A/B)×((C/D)/(E/F))

A/B=relative gMFI

A: gMFI of T2 cells with the addition of DMSO or peptide in the presence of antibody

B: gMFI of T2 cells with the addition of DMSO or peptide

(C/D)/(E/F)=corrected HLA/peptide complex count of T2 cells supplemented with DMSO or peptide.

C: gMFI of T2 cells with the addition of DMSO or peptide in the presence of HLA-A2 antibody.

D: gMFI of T2 cells with DMSO or peptide in the presence of mouse IgG2b antibody or peptide

E: gMFI of T2 cells supplemented with DMSO containing HLA-A2 antibody.

F: gMFI of T2 cells supplemented with DMSO containing mouse IgG2b antibody.

As shown in Fig. 1, anti-HLA/NY-ESO scFv: NYA-0001, 1143, 2044, 2045, 2047, 2048, 2060 and 2061 exhibited binding affinity to T2 cells supplemented with peptides containing point mutations introduced at amino acids 1, 4, 5 and 7 decreased by half or less compared with the wild-type NY-ESO peptide. This indicates that such scFv recognizes amino acids 1, 4, 5 and 7 of the NY-ESO peptide. This also indicates that NYA-1154 recognizes amino acids 1 and 5, NYA-1163 recognizes amino acids 1, 3, 4, 5, 6, and 7, NYA-2023, 2027, and 2035 recognize amino acids 1, 4, and 5, and NYA-2143 recognizes amino acids 1, 5, and 7. This also indicates that NYC-0003 and 0004 recognize amino acids 4 and 5.

(Example 6) Evaluation of antigen-binding specificity of anti-HLA/NY-ESO scFv

To search for a human peptide having an amino acid sequence similar to but not identical to that of the NY-ESO peptide: SLLMWITQC (SEQ ID NO:1) to which the antibody can bind (hereinafter referred to as a "homologous peptide"), a 9-mer peptide containing amino acids 1, 4, and 5, which are consistent with the peptides recognized by the antibody, was searched among human proteomes (Swiss-Prot). The target 9-mer peptide was analyzed for binding affinity to HLA-A0201 using NetMHCPan2.8, and its IC50 was predicted to be 500 nM or less. The 9-mer peptide shown in Fig. 2A was selected as a homologous peptide to evaluate the binding specificity of anti-HLA/NY-ESO scFv. The T2 cell concentration was adjusted to an adequate level in AIM-V medium (Thermo Fisher Scientific) containing 20% FBS, NY-ESO peptide solution (SEQ ID NO:1), DOLPP1, IL20RB, PRKD2, CD163 and P2RY8 homologous peptides (SEQ ID NO:131 (Fig. 132), SEQ ID NO:132 (Fig. 133), SEQ ID NO:133 (Fig. 134), SEQ ID NO:134 (Fig. 135) and SEQ ID NO:135 (Fig. 136)), or gp100 peptide (SEQ ID NO:130 (Fig. 131)) (Sigma Genosys) dissolved to 5 mM in DMSO was added to a final concentration of 50 μM, or DMSO was added in an amount 1/100, and the binding affinity of the antibodies was estimated in the same manner as in Example 5. The standardized gMFI indicating the binding affinity of each scFv standardized with the amount of HLA/peptide complex on T2 cells was determined according to the following equation.

Standardized gMFI=(A/B)×((C/D)/(E/F))

A/B=relative gMFI

A: gMFI of T2 cells with the addition of DMSO or peptide in the presence of antibody

B: gMFI of T2 cells with the addition of DMSO or peptide

(C/D)/(E/F)=corrected HLA/peptide complex count of T2 cells supplemented with DMSO or peptide.

C: gMFI of T2 cells supplemented with DMSO containing HLA-A2 antibody or peptide.

D: gMFI of T2 cells with the addition of DMSO or peptide in the presence of mouse IgG2b antibody.

E: gMFI of T2 cells supplemented with DMSO containing HLA-A2 antibody.

F: gMFI of T2 cells supplemented with DMSO containing mouse IgG2b antibody.

As shown in Fig. 2B, anti-HLA/NY-ESO scFv: NYA-0001, 1143, 1163, 2023, 2027, 2035, 2044, 2045, 2047, 2048, 2060, 2061 and 2143 did not bind to T2 cells complemented with any homologous peptides. This indicates that such scFv has high specificity. In contrast, NYA-1154, NYC-0003 and 0004 were found to bind to T2 cells complemented with some homologous peptides.

(Example 7) Preparation of Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecule

7)-1 Preparation of Fc-conjugated expression vector for bispecific anti-HLA/NY-ESO-anti-CD3 molecule

7)-1-1 Obtaining an expression vector for the bispecific TaFv-Fc type heterodimer molecule

To evaluate the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc heterodimer molecules, expression vectors for various molecules were constructed. The anti-HLA/NY-ESO antibodies used were NYA-1143, NYA-2143, NYA-1163, NYA-2023, NYA-2027, NYA-2035, NYA-2044, NYA-2045, NYA-2047, NYA-2048, NYA-2060, and NYA-2061. The anti-CD3 antibody used was a humanized anti-CD3 scFv; i.e., C3E-7085 (WO 2018/117237). As a heterodimeric Fc sequence, an Fc sequence containing a mutation introduced into it to reduce effector functions and form a heteropolymer is used (WO 2014/190441).

A DNA fragment encoding Fc (HC1 or HC2) containing a mutation introduced into it to reduce effector functions and form a heteropolymer was synthesized (Fasmac Co., Ltd.), an expression vector for mammalian cells containing pcDNA3.3 (Thermo Fisher Scientific) as a backbone was prepared using an In-Fusion HD Cloning Kit (CLONTECH), and the resulting vector was designated as “p_HC1”.

A mammalian cell expression vector was prepared containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of a taFv containing NYA-1143 ligated to C3E-7085 via a GGGGS linker and designated "p_NYF-0016-HC2".

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2143 ligated to C3E-7085 via a GGGGS linker was prepared by site-directed mutagenesis in p_NYF-0016-HC2 and designated as “p_NYF-0019-HC2”.

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-1163 ligated to C3E-7085 using a GGGGS linker was prepared by replacing the nucleotide sequence encoding NYA-1143 of p_NYF-0016-HC2 with a DNA fragment encoding NYA-1163 using the In-Fusion HD Cloning Kit (CLONTECH) and designated as “p_NYF-0022-HC2”.

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2023 ligated to C3E-7085 via a GGGGS linker was prepared by site-directed mutagenesis in p_NYF-0016-HC2 and designated as “p_NYF-0023-HC2”.

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of a taFv containing NYA-2027 ligated to C3E-7085 via a GGGGS linker was prepared by site-directed mutagenesis of a mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of a taFv containing NYA-0001 ligated to C3E-7085 via a GGGGS linker and designated as "p_NYF-0027-HC2".

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2035 ligated to C3E-7085 via a GGGGS linker was prepared by site-directed mutagenesis in p_NYF-0016-HC2 and designated as “p_NYF-0035-HC2”.

An expression vector for mammalian cells containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2044 ligated to C3E-7085 using a GGGGS linker was prepared by replacing the nucleotide sequence encoding NYA-1143 of p_NYF-0016-HC2 with a DNA fragment encoding NYA-2044 using the In-Fusion HD Cloning Kit (CLONTECH) and designated as “p_NYF-0044-HC2”.

An expression vector for mammalian cells containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2045 ligated to C3E-7085 using a GGGGS linker was prepared by replacing the nucleotide sequence encoding NYA-1143 of p_NYF-0016-HC2 with a DNA fragment encoding NYA-2045 using the In-Fusion HD Cloning Kit (CLONTECH) and designated as “p_NYF-0045-HC2”.

An expression vector for mammalian cells containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2047 ligated to C3E-7085 using a GGGGS linker was prepared by replacing the nucleotide sequence encoding NYA-1143 of p_NYF-0016-HC2 with a DNA fragment encoding NYA-2047 using the In-Fusion HD Cloning Kit (CLONTECH) and designated as “p_NYF-0047-HC2”.

An expression vector for mammalian cells containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2048 ligated to C3E-7085 using a GGGGS linker was prepared by replacing the nucleotide sequence encoding NYA-1143 of p_NYF-0016-HC2 with a DNA fragment encoding NYA-2048 using the In-Fusion HD Cloning Kit (CLONTECH) and designated as “p_NYF-0048-HC2”.

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2060 ligated to C3E-7085 via a GGGGS linker was prepared by site-directed mutagenesis in p_NYF-0044-HC2 and designated as “p_NYF-0060-HC2”.

A mammalian cell expression vector containing a DNA fragment encoding HC2 integrated into the carboxyl terminus of taFv containing NYA-2061 ligated to C3E-7085 via a GGGGS linker was prepared by site-directed mutagenesis in p_NYF-0045-HC2 and designated as “p_NYF-0061-HC2”.

The nucleotide sequence of p_HC1 was reanalyzed and the nucleotide sequence of full-length HC1 was found to be the nucleotide sequence shown in SEQ ID NO:71 of the sequence listing (Fig. 72).

The nucleotide sequence of p_NYF-0016-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0016-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:72 of the sequence listing (Fig. 73).

The nucleotide sequence of p_NYF-0019-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0019-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:73 of the sequence listing (Fig. 74).

The nucleotide sequence of p_NYF-0022-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0022-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:74 of the sequence listing (Fig. 75).

The nucleotide sequence of p_NYF-0023-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0023-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:75 of the sequence listing (Fig. 76).

The nucleotide sequence of p_NYF-0027-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0027-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:76 of the sequence listing (Fig. 77).

The nucleotide sequence of p_NYF-0035-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0035-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:77 of the sequence listing (Fig. 78).

The nucleotide sequence of p_NYF-0044-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0044-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:78 of the sequence listing (Fig. 79).

The nucleotide sequence of p_NYF-0045-HC2 was reanalyzed and the nucleotide sequence of full-length NYF-0045-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:79 of the sequence listing (Fig. 80).

The nucleotide sequence of p_NYF-0047-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0047-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:80 of the sequence listing (Fig. 81).

The nucleotide sequence of p_NYF-0048-HC2 was reanalyzed and the nucleotide sequence of the full-length NYF-0048-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:81 of the sequence listing (Fig. 82).

The nucleotide sequence of p_NYF-0060-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0060-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:82 of the sequence listing (Fig. 83).

The nucleotide sequence of p_NYF-0061-HC2 was reanalyzed, and the nucleotide sequence of the full-length NYF-0061-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:83 of the sequence listing (Fig. 84).

Based on the above nucleotide sequences, the amino acid sequences of full-length HC1, NYF-0016-HC2, NYF-0019-HC2, NYF-0022-HC2, NYF-0023-HC2, NYF-0027-HC2, NYF-0035-HC2, NYF-0044-HC2, NYF-0045-HC2, NYF-0047-HC2, NYF-0048-HC2, NYF-0060-HC2 and NYF-0061-HC2 encoded in this manner were identified.

The amino acid sequence of full-length HC1 is the amino acid sequence shown in SEQ ID NO:84 of the sequence listing (Fig. 85). In such an amino acid sequence, amino acids 1-19 constitute the signal sequence, and amino acids 20-246 constitute the HC.

The amino acid sequence of full-length NYF-0016-HC2 is shown in SEQ ID NO:85 of the sequence listing (Fig. 86). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-1143-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0019-HC2 is shown in SEQ ID NO:86 of the sequence listing (Fig. 87). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2143-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0022-HC2 is shown in SEQ ID NO:87 of the sequence listing (Fig. 88). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-1163-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of the full-length NYF-0023-HC2 is shown in SEQ ID NO:88 of the sequence listing (Fig. 89). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2023-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0027-HC2 is shown in SEQ ID NO:89 of the sequence listing (Fig. 90). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2027-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0035-HC2 is shown in SEQ ID NO:90 of the sequence listing (Fig. 91). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2035-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0044-HC2 is shown in SEQ ID NO:91 of the sequence listing (Fig. 92). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2044-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0045-HC2 is shown in SEQ ID NO:92 of the sequence listing (Fig. 93). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2045-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0047-HC2 is shown in SEQ ID NO:93 of the sequence listing (Fig. 94). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2047-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0048-HC2 is shown in SEQ ID NO:94 of the sequence listing (Fig. 95). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2048-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of the full-length NYF-0060-HC2 is shown in SEQ ID NO:95 of the sequence listing (Fig. 96). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2060-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of full-length NYF-0061-HC2 is shown in SEQ ID NO:96 of the sequence listing (Fig. 97). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2061-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

7)-1-2 Obtaining an expression vector for the bispecific taFv-Fab-Fc type heterodimer molecule

An expression vector for a bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fab-Fc-type heterodimer molecule is constructed. NYA-0001 is used as the anti-HLA/NY-ESO antibody. A humanized anti-CD3 scFv is used as the anti-CD3 antibody; that is, C3E-7085 (WO 2018/117237). An Fc sequence containing a mutation introduced into it to reduce effector functions and form a heteropolymer is used as the Fc heterodimeric sequence (WO 2014/190441).

An expression vector for mammalian cells containing DNA fragments encoding the variable region of the heavy chain of NYA-0001, the CH1 region derived from human IgG, and the region containing the mutation that reduces the effector functions and encodes the HC1-k deletion integrated therein is prepared and designated as "p_NYA-0001-Fab-HC1-k deletion". In addition, an expression vector for mammalian cells containing DNA fragments encoding the variable region of the light chain of NYA-0001 and the CL region derived from human IgG integrated therein is prepared and designated as "p_NYA-0001-LC".

The nucleotide sequence of the p_NYA-0001-Fab-HC1-k deletion was reanalyzed, and the nucleotide sequence of the full-length NYA-0001-Fab-HC1-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:97 (Fig. 98) of the Sequence Listing.

The nucleotide sequence of p_NYA-0001-LC was reanalyzed, and the nucleotide sequence of full-length NYA-0001-LC was found to be the nucleotide sequence shown in SEQ ID NO:98 (Fig. 99) of the sequence listing.

Based on the above nucleotide sequences, the amino acid sequences of the full-length deletion encoded by them, NYA-0001-Fab-HC1-k and NYA-0001-LC, are identified.

The amino acid sequence of the full-length NYA-0001-Fab-HC1-k deletion is the amino acid sequence shown in SEQ ID NO:99 of the sequence listing (Fig. 100). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 20-139 constitute a variable region, and amino acids 140-468 constitute a constant region. In addition, amino acids 45-52 constitute CDRH1 (SEQ ID NO:54 (Fig. 61)), amino acids 70-77 constitute CDRH2 (SEQ ID NO:55 (Fig. 61)), and amino acids 116-128 constitute CDRH3 (SEQ ID NO:56 (Fig. 61)).

The amino acid sequence of full-length NYA-0001-LC is shown in SEQ ID NO:100 of the sequence listing (Fig. 101). In such amino acid sequence, amino acids 1-20 constitute a signal sequence, amino acids 21-131 constitute a variable region, and amino acids 132-237 constitute a constant region. In addition, amino acids 46-53 constitute CDRL1 (SEQ ID NO:57 (Fig. 61)), amino acids 71-73 constitute CDRL2 (SEQ ID NO:58 (Fig. 61)), and amino acids 110-121 constitute CDRL3. (SEQ ID NO:59 (Fig. 61)).

7)-2 Expression of Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecule

7)-2-1 Expression of a bispecific taFv-Fc type heterodimer molecule

Expi293F cells (Thermo Fisher Scientific) were subcultured according to the instructions. Expi293F cells in logarithmic growth phase were diluted to 2.5× ¹⁰⁶ cells/mL in Expi293 Expression Medium (Thermo Fisher Scientific) and used to produce various bispecific molecules. A mixture (0.3 mg) containing p_NYF-0016-HC2 and p_HC1 vector at a ratio of 1:1.5 and 0.9 mg polyethyleneimine (Polyscience #24765) was added to 20 mL of Opti-Pro SFM medium (Thermo Fisher Scientific). The mixture was gently mixed, left for five minutes, and then added to Expi293F cells. The culture supernatant obtained by shaking the culture in an incubator at 37°C in the presence of 8% _CO2 at 135 rpm for 6 days was filtered through a 0.2 μm filter (Millipore). Thus, the culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0016) was obtained. The amino acid sequences obtained by the expression vectors constituting NYF-0016 are shown in SEQ ID NO: 85 of the sequence listing (Fig. 86) and in SEQ ID NO: 84 of the sequence listing (Fig. 85).

In the same manner, the culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc-type heterodimer molecule (NYF-0019) was prepared using p_NYF-0019-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0019 are shown in SEQ ID NO:86 of the sequence listing (Fig. 87) and in SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc-type heterodimer molecule (NYF-0022) was prepared using p_NYF-0022-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0022 are shown in SEQ ID NO:87 of the sequence listing (Fig. 88) and in SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc-type heterodimer molecule (NYF-0023) was prepared using p_NYF-0023-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0023 are shown in SEQ ID NO:88 of the sequence listing (Fig. 89) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0027) was prepared using p_NYF-0027-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0027 are shown in SEQ ID NO:89 of the sequence listing (Fig. 90) and in SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0035) was prepared using p_NYF-0035-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0035 are shown in SEQ ID NO:90 of the sequence listing (Fig. 91) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0044) was prepared using p_NYF-0044-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0044 are shown in SEQ ID NO:91 of the sequence listing (Fig. 92) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc-type heterodimer molecule (NYF-0045) was prepared using p_NYF-0045-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0045 are shown in SEQ ID NO:92 of the sequence listing (Fig. 93) and in SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0047) was prepared using p_NYF-0047-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0047 are shown in SEQ ID NO:93 of the sequence listing (Fig. 94) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0048) was prepared using p_NYF-0048-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0048 are shown in SEQ ID NO:94 of the sequence listing (Fig. 95) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0060) was prepared using p_NYF-0060-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0060 are shown in SEQ ID NO:95 of the sequence listing (Fig. 96) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYF-0061) was prepared using p_NYF-0061-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYF-0061 are shown in SEQ ID NO:96 of the sequence listing (Fig. 97) and in SEQ ID NO:84 of the sequence listing (Fig. 85).

7)-2-2 Expression of bispecific taFv-Fab-heterodimer Fc type molecules

In the same manner as in 7)-2-1, the culture supernatant of the taFv-Fab-Fc type heterodimer bispecific molecule (NYF-0058) was prepared using a vector mixture containing p_NYF-0023-HC2, p_NYA-0001-Fab-HC1-k deletion, and p_NYA-0001-LC at a ratio of 1:1:1.5. The amino acid sequences obtained by expression of the vectors constituting NYF-0058 are represented by amino acids 20-745 of SEQ ID NO: 88 of the sequence listing (Fig. 89), amino acids 20-468 of SEQ ID NO: 99 of the sequence listing (Fig. 100), and amino acids 21-237 of SEQ ID NO: 100 of the sequence listing (Fig. 101).

7)-3 Purification of Fc-conjugated bispecific anti-HLA/NY-ESO-anti-CD3 molecule

The various bispecific molecules were purified from the culture supernatants obtained in 7)-2 by two steps of protein A affinity chromatography and gel filtration chromatography.

The culture supernatant was applied to a MabSelectSuRe column equilibrated with PBS at pH 7.4 (GE Healthcare Bioscience, also referred to simply as “GE Healthcare”) to allow the bispecific target molecules to adsorb to it. After removing non-adsorbed components with PBS, the adsorbed components were eluted with acetate buffer (pH 3.5). The eluted fraction was neutralized with Tris buffer (pH 9.5), concentrated, and then applied to a Superdex 200 10/300 gel filtration column (GE Healthcare Bioscience) equilibrated with 25 mM histidine, 300 mM NaCl, 5% sorbitan at pH 5.5 in advance. From the peak fraction obtained by gel filtration chromatography, fractions equivalent to the target heterodimer were collected and the formation of the bispecific anti-HLA/NY-ESO-anti-CD3 target molecule was confirmed by SDS-polyacrylamide electrophoresis (SDS-PAGE). Purified protein samples were subjected to analytical SEC, the degree of purification and concentration were determined, and then the samples were subjected to various types of assessment.

(Example 8) Evaluation of the cytotoxicity of an Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule

8)-1 Obtaining target cells

Endogenous human NY-ESO expressing cells (U266B1 and NCI-H1703) and endogenous human NY-ESO non-expressing cells (AGS and CFPAC-1) were adjusted to 1× ¹⁰⁶ cells/ml in RPMI 1640 medium containing 10% FBS (FUJIFILM Wako Pure Chemical Corporation), 100 μl of chromium-51 radionuclide (PerkinElmer) were added per 1 ml of each cell suspension, and the resulting products were cultured at 37°C in the presence of 5% _CO2 for 2 h. The cells were washed 2 times in RPMI 1640 medium containing 10% FBS, resuspended to 1× ¹⁰⁵ cells/ml in RPMI 1640 medium containing 10% FBS, and then used as target cells.

8)-2 Obtaining effector cells

Commercially available frozen human PBMCs (Cellular Technology Limited) were thawed at 37°C, transferred to a solution containing RPMI 1640 medium containing 10% FBS supplemented with Anti-aggregate Wash reagent (Cellular Technology Limited), washed 2 times and adjusted to 1× ¹⁰⁶ cells/ml in RPMI 1640 medium containing 10% FBS to obtain effector cells.

8)-3 Cytotoxicity assay

The target cells obtained in 8)-1 were seeded in a 96-well U-bottom microplate at 50 μl/well. Various Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules obtained in Example 7 adjusted to different concentrations were added thereto at 50 μl/well, the effector cells obtained in Example 8)-2 were added thereto at 100 μl/well, centrifuged at room temperature and 1000 rpm for 1 minute, and then incubated at 37°C in the presence of 5% CO.₂for 20-24 hours. The supernatant (50 µl) is collected on a LumaPlate (PerkinElmer), dried at 50°C for approximately 2 hours, and then analyzed using a plate reader (TopCount, PerkinElmer). The test is performed in triplicate, and the rate of cell lysis is determined according to the following equation.

Cell lysis rate (%) = (A - B) / (C - B) × 100

A: number of sample wells

B: Average background value (wells without added antibodies) (n=3).

When the antibody is added, 50 µl of assay medium is added. The remaining procedures are the same as for the sample wells.

C: mean maximum release value (wells in which target cells are lysed by surfactant) (n=3).

When the antibody is added, 50 µl of assay medium is added. Surfactant is added at 100 µl, a 50 µl fraction is transferred to the LumaPlate as for the sample wells, and assays are performed.

As shown in Fig. 4A to Fig. 4F, different anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules exhibited cytotoxicity against endogenous human NY-ESO-expressing cells (U266B1 and NCI-H1703). The _EC50 calculation results obtained using the four-parameter logistic curve equation of the analytical software (Sigmaplot, version 12.0) for U266B1 cells are shown in Table 2, and the results for NCI-H1703 cells are shown in Table 3. “Not applicable (NA)” indicates that the curve could not be fitted because the correlation coefficient (R) was not calculated. In contrast, as shown in Fig. 4G to 4L, no cytotoxicity was observed on endogenous human NY-ESO-non-expressing cells (AGS and CFPAC-1).

[Table 2]

Name EC ₅₀ (nM) 1 NYF-0016 0.30 2 NYF-0019 0.20 3 NYF-0022 1.30 4 NYF-0023 0.90 5 NYF-0027 2.68 6 NYF-0035 0.97 7 NYF-0044 0.10 8 NYF-0045 0.18 9 NYF-0047 0.93 10 NYF-0048 1.10 11 NYF-0058 0.95 12 NYF-0060 0.39 13 NYF-0061 0.40

[Table 3]

Name EC ₅₀ (nM) 1 NYF-0016 0.90 2 NYF-0019 4.28 3 NYF-0022 NA* 4 NYF-0023 1.16 5 NYF-0027 6.40 6 NYF-0035 2.10 7 NYF-0044 1.11 8 NYF-0045 5.76 9 NYF-0047 7.63 10 NYF-0048 4.21 11 NYF-0058 1.03 12 NYF-0060 0.74 13 NYF-0061 1.04 *NA=not applicable

(Example 9) In vivo evaluation of the activity of Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules in PBMC-transfected human models

Human squamous cell lung cancer cell lines NCI-H1703 (ATCC) were adjusted to 6× ¹⁰⁷ cells/mL in PBS containing 50% Matrigel (Corning), and 0.1 mL of this preparation was injected subcutaneously into NOG mice (female, 6-7 weeks old) (day 0). On day 4, human PBMCs were adjusted to 3.75× ¹⁰⁷ cells/mL in PBS, and 0.2 mL of this compound was injected intravenously. Approximately 1 week thereafter (day 6 to 7), the measurement of the major diameter (mm) and minor diameter (mm) of the tumor was started using an electronic digital caliper over time, and the estimated tumor volume was calculated according to the following equation.

Estimated tumor volume ( ^mm3 ) = average estimated tumor volume among individuals

Estimated tumor volume in humans = major diameter x [minor diameter] ^2/ 2

On day 14, the mice were divided into groups of 5 or 6 mice each depending on the tumor volume, and various anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules (1 mg/kg, 1.5 mg/kg NYF-0058 for comparison with the same molecular weight) were administered intravenously. The administration was performed on days 14, 21, and 28. The antitumor effects were observed in the treatment groups administered with different anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules (Fig. 5A to Fig. 5C). The tumor growth inhibition (%) from day 31 to day 32 was calculated according to the following equation and shown in Table 4.

Tumor growth inhibition (%) = 100 - (estimated tumor volume in the treatment group/estimated tumor volume in the control group × 100)

[Table 4]

Name Tumor growth inhibition (%) 1 NYF-0016 100,0 2 NYF-0019 98.8 3 NYF-0022 90.3 4 NYF-0023 98.6 5 NYF-0027 99.1 6 NYF-0035 92.5 7 NYF-0044 100,0 8 NYF-0045 95.4 9 NYF-0047 78.9 10 NYF-0048 96.3 11 NYF-0058 88.5 12 NYF-0060 88.8 13 NYF-0061 100,0

(Example 10) Preparation of various Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecules

10)-1 Preparation of various expression vectors of Fc-conjugated bispecific anti-HLA/NY-ESO-anti-CD3 molecules.

10)-1-1 Obtaining an expression vector for a hybrid type bispecific molecule

To evaluate the bispecific molecules of the anti-HLA/NY-ESO-anti-CD3 hybrid type, expression vectors for various molecules are constructed. NYA-1143 is used as the anti-HLA/NY-ESO antibody. A humanized anti-CD3 scFv is used as the anti-CD3 antibody; i.e., C3E-7085 is used (PCT/JP2017/046006). An Fc sequence containing a mutation introduced into it to reduce effector functions and form a heteropolymer is used as the heterodimeric Fc sequence (WO 2014/190441).

An expression vector for mammalian cells containing DNA fragments encoding the variable region of the heavy chain of NYA-1143, the CH1 region derived from human IgG, and the Fc region in which a mutation was introduced to reduce effector functions and form a heteropolymer was obtained, and designated as "p_NYA-1143-Fab-HC1-k deletion". In addition, an expression vector for mammalian cells containing DNA fragments encoding the variable region of the light chain of NYA-1143 and the CL region derived from human IgG was obtained, and designated as "p_NYA-1143-LC". In addition, a mammalian cell expression vector containing a DNA fragment encoding an Fc region containing a mutation that reduces effector functions and forms a heteropolymer introduced into the carboxyl terminus of a humanized anti-CD3 scFv (C3E-7085) was prepared and designated as “p_C3E-7085-HC2-k deletion”.

The nucleotide sequence of the p_NYA-1143-Fab-HC1-k deletion was reanalyzed, and the nucleotide sequence of the full-length NYA-1143-Fab-HC1-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:101 of the sequence listing (Fig. 102).

The nucleotide sequence of p_NYA-1143-LC was reanalyzed and the nucleotide sequence of full-length NYA-1143-LC was found to be the nucleotide sequence shown in SEQ ID NO:102 of the sequence listing (Fig. 103).

The nucleotide sequence of the p_C3E-7085-HC2-k deletion was reanalyzed, and the nucleotide sequence of the full-length C3E-7085-HC2-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:103 of the sequence listing (Fig. 104).

Based on the above nucleotide sequences, the amino acid sequences of the full-length deletions encoded by them NYA-1143-Fab-HC1-k, NYA-1143-LC and C3E-7085-HC2-k are identified.

The amino acid sequence of the full-length NYA-1143-Fab-HC1-k deletion is the amino acid sequence shown in SEQ ID NO: 104 of the sequence listing (Fig. 105). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-139 constitute a variable region, and amino acids 140-468 constitute a constant region. In addition, amino acids 45-52 constitute CDRH1 (SEQ ID NO: 54 (Fig. 61)), amino acids 70-77 constitute CDRH2 (SEQ ID NO: 55 (Fig. 61)), amino acids 116-128 constitute CDRH3 (SEQ ID NO: 56 (Fig. 61)).

The amino acid sequence of the full-length NYA-1143-LC is the amino acid sequence shown in SEQ ID NO:105 of the sequence listing (Fig. 106). In such amino acid sequence, amino acids 1-20 constitute a signal sequence, amino acids 21-131 constitute a variable region, and amino acids 132-237 constitute a constant region. In addition, amino acids 46-53 constitute CDRL1 (SEQ ID NO:60 (Fig. 62)), amino acids 71-73 constitute CDRL2 (SEQ ID NO:58 (Fig. 61)), and amino acids 110-121 constitute CDRL3 (SEQ ID NO:59 (Fig. 61)).

The amino acid sequence of the full-length deletion of C3E-7085-HC2-k is the amino acid sequence shown in SEQ ID NO:106 of the sequence listing (Fig. 107). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-260 constitute C3E-7085, amino acids 261-262 constitute a linker, and amino acids 263-493 constitute an Fc containing a mutation introduced therein to reduce effector functions and form a heterodimer.

10)-1-2 Obtaining a vector for the expression of a bispecific molecule of a dual type

To evaluate the bispecific molecules of the dual anti-HLA/NY-ESO-anti-CD3 type, an expression vector was constructed, which is described less. An expression vector for mammalian cells was prepared containing a DNA fragment encoding the Fc region containing a mutation that reduces effector functions and forms a heteropolymer integrated into the carboxyl terminus of NYA-1143 and designated as "p_NYA-1143-HC1-k deletion".

The nucleotide sequence of the p_NYA-1143-HC1-k deletion was reanalyzed, and the nucleotide sequence of the full-length NYA-1143-HC1-k deletion was found to be the nucleotide sequence shown in SEQ ID NO: 107 of the sequence listing (Fig. 108). Based on the above nucleotide sequences, the amino acid sequence of the encoded full-length NYA-1143-HC1-k deletion was identified. The amino acid sequence of the full-length NYA-1143-HC1-k deletion was the amino acid sequence shown in SEQ ID NO: 108 (Fig. 109) of the sequence listing. In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-266 constitute NYA-1143, amino acids 267-268 constitute the linker, and amino acids 269-499 constitute the Fc, which contains a mutation introduced into it to reduce effector functions and form a heterodimer.

10)-1-3 Obtaining an expression vector for the bispecific scFv-Fab-Fc type heterodimer molecule

To evaluate bispecific molecules of anti-HLA/NY-ESO-anti-CD3 scFv-Fab-Fc type heterodimer, expression vectors for different molecules are constructed. NYA-1154 is used as anti-HLA/NY-ESO antibody, and C3E-7085 is used as anti-CD3 antibody. As heterodimeric Fc sequence, an Fc sequence containing a mutation introduced into it to reduce effector functions and form a heteropolymer is used.

A mammalian cell expression vector containing DNA fragments encoding the variable region of the heavy chain of NYA-1154, the CH1 region derived from human IgG, and the Fc region containing a mutation reducing effector functions and forming a heteropolymer integrated into the carboxyl terminus of C3E-7085 was prepared and designated as "p_C3E-7085-NYA-1154-Fab-HC2-k deletion". In addition, a mammalian cell expression vector containing DNA fragments encoding the variable region of the light chain of NYA-1154 and the CL region derived from human IgG integrated therein was prepared and designated as "p_NYA-1154-LC". In addition, a mammalian cell expression vector containing a DNA fragment encoding the HC1-k deletion integrated therein was prepared and designated as "p_OAA-HC1-k deletion".

The nucleotide sequence of the p_C3E-7085-NYA-1154-Fab-HC2-k deletion was reanalyzed, and the nucleotide sequence of the full-length C3E-7085-NYA-1154-Fab-HC2-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:109 of the sequence listing (Fig. 110).

The nucleotide sequence of p_NYA-1154-LC was reanalyzed and the nucleotide sequence of full-length NYA-1154-LC was found to be the nucleotide sequence shown in SEQ ID NO:110 (Fig. 111) of the sequence listing.

The nucleotide sequence of the p_OAA-HC1-k deletion was reanalyzed, and the nucleotide sequence of the full-length OAA-HC1-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:111 of the sequence listing (Fig. 112).

Based on the above nucleotide sequences, the amino acid sequences of the full-length deletions C3E-7085-NYA-1154-Fab-HC2-k, NYA-1154-LC and OAA-HC1-k encoded by them are identified.

The amino acid sequence of the full-length deletion of C3E-7085-NYA-1154-Fab-HC2-k is the amino acid sequence shown in SEQ ID NO: 112 of the sequence listing (Fig. 113). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-260 constitute C3E-7085, amino acids 261-385 constitute a linker, and amino acids 266-385 constitute the variable region of the heavy chain of NYA-1154. In addition, amino acids 386-714 constitute the deletion of HC2-k.

The amino acid sequence of the full-length NYA-1154-LC is the amino acid sequence shown in SEQ ID NO:113 of the sequence listing (Fig. 114). In such amino acid sequence, amino acids 1-20 constitute a signal sequence, amino acids 21-131 constitute a variable region, and amino acids 132-237 constitute a constant region. In addition, amino acids 46-53 constitute CDRL1 (SEQ ID NO:57 (Fig. 61)), amino acids 71-73 constitute CDRL2 (SEQ ID NO:58 (Fig. 61)), and amino acids 110-121 constitute CDRL3. (SEQ ID NO:63 (Fig. 64)).

The amino acid sequence of the full-length OAA-HC1-k deletion is the amino acid sequence shown in SEQ ID NO:114 of the sequence listing (Fig. 115). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, and amino acids 20-245 constitute an OAA-HC1-k deletion.

10)-1-4 Generation of expression vectors of bispecific TaFv-Fc type heterodimer molecules for evaluation of different formats

In order to evaluate bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecules for evaluation of different formats, expression vectors for different molecules are constructed. NYA-1143 and NYA-1154 are used as anti-HLA/NY-ESO antibodies. Humanized anti-CD3 scFv is used as anti-CD3 antibody; i.e., C3E-7085 is used. As heterodimeric Fc sequence, an Fc sequence containing a mutation introduced into it to reduce effector functions and form a heteropolymer is used.

A mammalian cell expression vector containing a DNA fragment encoding the HC2-k deletion integrated into the carboxyl terminus of taFv containing NYA-1154 ligated to C3E-7085 via a GGGGS linker was prepared and designated as “p_NYF-0010-HC2-k deletion”.

A mammalian cell expression vector containing a DNA fragment encoding the HC2-k deletion integrated into the carboxyl terminus of a taFv containing C3E-7085 ligated to NYA-1154 via a GGGGS linker was prepared and designated as “p_NYF-0004-HC2-k deletion”.

A mammalian cell expression vector containing a DNA fragment encoding the HC2-k deletion integrated into the carboxyl terminus of a taFv containing NYA-1143 ligated to C3E-7085 via a GGGGS linker was prepared and designated as “p_NYF-0011-HC2-k deletion”.

The nucleotide sequence of the p_NYF-0010-HC2-k deletion was reanalyzed, and the nucleotide sequence of the full-length NYF-0010-HC2-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:115 of the sequence listing (Fig. 116).

The nucleotide sequence of the p_NYF-0004-HC2-k deletion was reanalyzed, and the nucleotide sequence of the full-length NYF-0004-HC2-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:116 of the sequence listing (Fig. 117).

The nucleotide sequence of the p_NYF-0011-HC2-k deletion was reanalyzed, and the nucleotide sequence of the full-length NYF-0011-HC2-k deletion was found to be the nucleotide sequence shown in SEQ ID NO:117 of the sequence listing (Fig. 118).

Based on the above nucleotide sequences, the amino acid sequences of the full-length NYF-0010-HC2-k deletion, NYF-0004-HC2-k deletion, and NYF-0011-HC2-k deletion encoded by them were identified.

The amino acid sequence of the full-length NYF-0010-HC2-k deletion is the amino acid sequence shown in SEQ ID NO:118 (Fig. 119) of the sequence listing. In such amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-1154-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-744 constitute the HC2-k deletion.

The amino acid sequence of the full-length NYF-0004-HC2-k deletion is the amino acid sequence shown in SEQ ID NO:119 (Fig. 120) of the sequence listing. In such amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute C3E-7085-NYA-1154 taFv, amino acids 512-513 constitute the linker, and amino acids 514-744 constitute the HC2-k deletion.

The amino acid sequence of the full-length NYF-0011-HC2-k deletion is the amino acid sequence shown in SEQ ID NO:120 of the sequence listing (Fig. 121). In such an amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-1143-C3E-7085 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute the HC2-k deletion.

10)-2 Expression of Fc-conjugated bispecific anti-HLA/NY-ESO-anti-CD3 molecules of different formats

10)-2-1 Expression of hybrid type bispecific molecules

In the same manner as in 7)-2-1, the culture supernatant of the bispecific hybrid-type molecule (NYG-3143) was obtained using a vector mixture containing p_NYA-1143-Fab-HC1-k deletion, p_C3E-7085-HC2-k deletion, and p_NYA-1143-LC at a ratio of 1:1:1.5. The amino acid sequences obtained by the expression vectors constituting NYG-3143 are shown in SEQ ID NO: 104 of the sequence listing (Fig. 105), SEQ ID NO: 105 of the sequence listing (Fig. 106), and SEQ ID NO: 106 of the sequence listing (Fig. 107).

10)-2-2 Expression of bispecific molecules of dual type

In the same manner as in 7)-2-1, the culture supernatant of the dual-type bispecific molecule (NYG-2143) was prepared using a vector mixture containing the deletion p_NYA-1143-HC1-k and the deletion p_C3E-7085-HC2-k at a ratio of 2:1. The amino acid sequences obtained by the expression vectors constituting NYG-2143 are shown in SEQ ID NO: 108 of the sequence listing (Fig. 109) and in SEQ ID NO: 106 of the sequence listing (Fig. 107).

10)-2-3 Expression of bispecific scFv-Fab-heterodimer Fc type molecules

In the same manner as in 7)-2-1, the culture supernatant of the bispecific scFv-Fab-Fc type heterodimer molecule (scFv-Fab-Fc type) (NYF-0003) was prepared using a vector mixture containing p_C3E-7085-NYA-1154-Fab-HC2-k deletion, p_OAA-HC1-k deletion and p_NYA-1154-LC at a ratio of 1:1:1.5. The amino acid sequences obtained by the expression vectors constituting NYF-0003 are shown in SEQ ID NO: 112 of the sequence listing (Fig. 113), SEQ ID NO: 113 of the sequence listing (Fig. 114) and SEQ ID NO: 114 of the sequence listing (Fig. 115).

10)-2-4 Expression of bispecific taFv-Fc type heterodimer molecules for evaluation of different formats

In the same manner as in 7)-2-1, the culture supernatant of the anti-HLA/NY-ESO-anti-CD3 taFv-Fc type bispecific molecule (taFv-Fc type) (NYF-0010) was prepared using p_NYF-0010-HC2-k deletion and p_OAA-HC1-k deletion. The amino acid sequences obtained by the expression vectors constituting NYF-0010 are shown in SEQ ID NO: 118 of the sequence listing (Fig. 119) and in SEQ ID NO: 114 of the sequence listing (Fig. 115).

The culture supernatant of the anti-HLA/NY-ESO-anti-CD3 (NYF-0004) taFv (inverted)-Fc type heterodimer bispecific molecule (taFv (inverted)-Fc type) anti-HLA/NY-ESO-anti-CD3 (NYF-0004) was prepared using p_NYF-0004-HC2-k deletion and p_OAA-HC1-k deletion. The amino acid sequences obtained by the expression vectors constituting NYF-0004 are shown in SEQ ID NO: 119 of the sequence listing (Fig. 120) and SEQ ID NO: 114 of the sequence listing (Fig. 115).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type molecule (NYF-0011) was prepared using p_NYF-0011-HC2-k deletion and p_OAA-HC1-k deletion. The amino acid sequences obtained by the expression vectors constituting NYF-0011 are shown in SEQ ID NO: 120 of the sequence listing (Fig. 121) and in SEQ ID NO: 114 of the sequence listing (Fig. 115).

10)-3 Purification of Fc-conjugated bispecific anti-HLA/NY-ESO-anti-CD3 molecules of different formats

The various bispecific molecules were purified from the culture supernatants obtained in 10)-2-1 and 10)-2-2 by two-step protein A affinity chromatography and ceramic hydroxyapatite chromatography.

The culture supernatant was applied to a MabSelectSuRe column equilibrated with PBS at pH 7.4 (GE Healthcare Bioscience) to allow the bispecific target molecules to adsorb to it. After removal of non-adsorbed components with PBS, the adsorbed components were eluted with acetate buffer (pH 3.6). The eluted fraction was neutralized with Tris buffer (pH 9.5) and the buffer was replaced with 25 mM histidine, 150 mM NaCl, 5% sorbitan at pH 5.5. The target fraction solution diluted 5-fold with a buffer containing 10 mM potassium phosphate and 50 mM MES (pH 6.5) was applied to a ceramic hydroxyapatite column (Bio-Scale CHT Type-1 Hydroxyapatite Column, BioRad Japan) equilibrated with 10 mM potassium phosphate and 50 mM MES (pH 6.5). Elution in a linear concentration gradient was carried out using sodium chloride and fractions equivalent to the target heterodimer were collected. The fractions were pre-applied to a Superdex 200 10/300 gel filtration column (GE Healthcare Bioscience) equilibrated with 25 mM histidine, 300 mM NaCl, 5% sorbitan at pH 6.0. From the peak fraction obtained by gel filtration chromatography, fractions equivalent to the target heterodimer are collected and the buffer is exchanged with 25 mM histidine, 300 mM NaCl, 5% sorbitan at pH 5.5. The formation of the bispecific anti-HLA/NY-ESO-anti-CD3 target molecule is confirmed by SDS-polyacrylamide electrophoresis (SDS-PAGE). Purified protein samples are subjected to analytical SEC, the degree of purification and concentration are determined, and then the samples are subjected to various types of assessment.

The various bispecific molecules were purified from the culture supernatants obtained in 10)-2-3 and 10)-2-4 by a 2-step protein A affinity chromatography and gel filtration chromatography in the same manner as in 7)-3. The formation of the bispecific anti-HLA/NY-ESO-anti-CD3 target molecule was confirmed by SDS-polyacrylamide electrophoresis (SDS-PAGE). Samples of the purified protein were subjected to analytical SEC, the degree of purification and concentration were determined, and then the samples were subjected to various types of assessment.

(Example 11) In vitro assessment of the activity of Fc-conjugated bispecific anti-HLA-A2/NY-ESO-anti-CD3 molecules of different formats

11)-1 In vitro assessment of the activity of bispecific molecules of anti-HLA-A2/NY-ESO-anti-CD3 hybrid type, dual type and taFv-Fc type

11)-1-1 Obtaining target cells

U266B1 obtained in the same manner as in Example 8)-1 were used as target cells.

11)-1-2 Obtaining effector cells

Commercially available frozen human PBMCs (Cellular Technology Limited), prepared according to the procedure of Example 8)-2, were used as effector cells.

11)-1-3 Cytotoxicity assay

The target cells obtained in Example 11)-1-1 are seeded in a 96-well U-bottom microplate at 50 μl/well. The anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules of various formats obtained in Example 10 and adjusted to different concentration levels are added thereto in an amount of 50 μl/well, the effector cells obtained in Example 11)-1-2 are added at 100 μl/well, centrifuged at room temperature and 1000 rpm for 1 minute, and then incubated at 37°C in the presence of 5% CO₂for 20-24 hours. The supernatant (50 µl) is collected on a LumaPlate (PerkinElmer), dried at 50°C for approximately 2 hours, and then analyzed using a plate reader (TopCount, PerkinElmer). The test is performed in triplicate, and the rate of cell lysis is determined according to the following equation.

Cell lysis rate (%) = (A - B) / (C - B) × 100

A: number of sample wells

B: Average background value (wells without added antibodies) (n=3).

When the antibody is added, 50 µl of assay medium is added. The remaining procedures are the same as for the sample wells.

C: mean maximum release value (wells in which target cells are lysed by surfactant) (n=3).

When the antibody was added, 50 μl of assay medium was added. The surfactant was added in an amount of 100 μl, a 50 μl fraction was transferred to the LumaPlate as in the case of the sample wells, and then the assays were carried out. As shown in Fig. 7A, the taFv-Fc type showed cytotoxicity. The EC ₅₀ calculation results obtained using the four-parameter logistic curve equation of the analysis software (Sigmaplot, version 12.0) are shown in Table 5. “Not applicable (NA)” means that the curve fitting could not be performed because the correlation coefficient (R) was not calculated. Neither the hybrid nor the dual type showed cytotoxicity.

[Table 5]

Format Name EC ₅₀ (nM) 1 Hybrid NYG-3143 NA* 2 Double NYG-2143 NA* 3 taFv-Fc NYF-0011 0.27 * NA=not applicable

11)-2 In vitro assessment of the activity of bispecific molecules anti-HLA-A2/NY-ESO-anti-CD3 scFv-Fab-Fc type, taFv-Fc type and taFv (inverted)-Fc type

11)-2-1 Obtaining target cells

U266B1 obtained in the same manner as in Example 8)-1 were used as target cells.

11)-2-2 Obtaining effector cells

Commercially available frozen human PBMCs (Cellular Technology Limited), prepared according to the procedure of Example 8)-2, were used as effector cells.

11)-2-3 Cytotoxicity assay

The target cells obtained in Example 11)-2-1 are seeded in a 96-well U-bottom microplate at 50 μl/well. The anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules of various formats obtained in Example 10 and adjusted to different concentration levels are added thereto in an amount of 50 μl/well, the effector cells obtained in Example 11)-1-2 are added at 100 μl/well, centrifuged at room temperature and 1000 rpm for 1 minute, and then incubated at 37°C in the presence of 5% CO₂for 20-24 hours. The supernatant (50 µl) is collected on a LumaPlate (PerkinElmer), dried at 50°C for approximately 2 hours, and then analyzed using a plate reader (TopCount, PerkinElmer). The test is performed in triplicate, and the rate of cell lysis is determined according to the following equation.

Cell lysis rate (%) = (A - B) / (C - B) × 100

A: number of sample wells

B: Average background value (wells without added antibodies) (n=3).

When the antibody is added, 50 µl of assay medium is added. The remaining procedures are the same as for the sample wells.

C: mean maximum release value (wells in which target cells are lysed by surfactant) (n=3).

When the antibody was added, 50 μl of the assay medium was added. The surfactant was added in an amount of 100 μl, a 50 μl fraction was transferred to the LumaPlate as in the case of the sample wells, and then the assays were carried out. As shown in Fig. 7B, the taFv-Fc type exhibited cytotoxicity. The calculation results of _EC50 obtained using the four-parameter logistic curve equation of the analytical software (Sigmaplot, version 12.0) are shown in Table 6. “Not applicable (NA)” means that the curve fitting could not be performed because the correlation coefficient (R) was not calculated. Compared with taFv-Fc type heterodimer, scFv-Fab-Fc type heterodimer and taFv (inverted)-Fc type heterodimer showed lower activity.

[Table 6]

Format Name EC ₅₀ (nM) 1 scFv-Fab-Fc heterodimer NYF-0003 NA* 2 taFv-Fc heterodimer NYF-0010 0.13 3 taFv (inverted) Fc heterodimer NYF-0004 0.88 *NA=not applicable

(Example 12) Preparation of Various Fc-Conjugated Anti-HLA/NY-ESO-Anti-CD3 Bispecific Molecules by Mutagenesis and Format Modification of NYF-0061

12)-1 Production of various Fc-conjugated expression vectors of bispecific anti-HLA/NY-ESO-anti-CD3 molecules by mutagenesis and modification of NYF-0061 format

To improve physical properties, NYF-0061 mutants are constructed and vectors are obtained that comprise the molecules described below.

A mutation is introduced into the C3E-7085 sequence of the p_NYF-0061-HC2 vector, which contains the nucleotide sequence encoding the amino acid sequence of NYF-0061, and the resulting gene is designated as “p_NYZ-0038-HC2”.

A mutation was introduced into the NYA-2061 sequence and the C3E-7085 sequence of the p_NYF-0061-HC2 vector, containing the nucleotide sequence encoding the amino acid sequence of NYF-0061, and the results were designated as “p_NYZ-0082-HC2” and “p_NYZ-0083-HC2,” respectively.

The nucleotide sequence of p_NYZ-0038-HC2 was reanalyzed and the nucleotide sequence of the full-length NYZ-0038-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:152 of the sequence listing (Fig. 148).

The nucleotide sequence of p_NYZ-0082-HC2 was reanalyzed and the nucleotide sequence of the full-length NYZ-0082-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:153 of the sequence listing (Fig. 149).

The nucleotide sequence of p_NYZ-0083-HC2 was reanalyzed and the nucleotide sequence of the full-length NYZ-0083-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:154 of the sequence listing (Fig. 150).

Based on the nucleotide sequences given above, the amino acid sequences of the full-length NYZ-0038-HC2, NYZ-0082-HC2 and NYZ-0083-HC2 encoded by them are identified.

The amino acid sequence of the full-length NYZ-0038-HC2 is the amino acid sequence shown in SEQ ID NO:155 of the sequence listing (Fig. 151). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-511 constitute the NYA-2061-C3E-7096 taFv, amino acids 512-513 constitute the linker, and amino acids 514-745 constitute HC2.

The amino acid sequence of the full-length NYZ-0082-HC2 is the amino acid sequence shown in SEQ ID NO:156 of the sequence listing (Fig. 152). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-516 constitute the NYA-3061-C3E-7096 taFv, amino acids 517-518 constitute the linker, and amino acids 519-750 constitute HC2.

The amino acid sequence of the full-length NYZ-0083-HC2 is the amino acid sequence shown in SEQ ID NO:157 of the sequence listing (Fig. 153). In this amino acid sequence, amino acids 1-19 constitute the signal sequence, amino acids 21-516 constitute the NYA-3061-C3E-7097 taFv, amino acids 517-518 constitute the linker, and amino acids 519-750 constitute HC2.

To study the modification of the NYF-0061 format in the scFv-Fab heterodimer of the Fc type, vectors were constructed that comprise the molecules described below.

A mammalian cell expression vector comprising DNA fragments encoding an amino acid sequence included in a polypeptide comprising (i) the heavy chain variable region of C3E-7085, (ii) the CH1 region derived from human IgG, and (iii) the Fc region containing a mutation to reduce effector functions and form a heteropolymer, integrated in that order into the carboxyl terminus of NYA-3061, is prepared and designated as "p_NYZ-1010-HC2". In addition, a mammalian cell expression vector comprising a DNA fragment encoding an amino acid sequence included in a polypeptide comprising the CL region derived from human IgG added to the carboxyl terminus of the light chain variable region of C3E-7085 is prepared and designated as "p_C3E-7085-LC".

The nucleotide sequence of p_NYZ-1010-HC2 was reanalyzed and the nucleotide sequence of the full-length NYZ-1010-HC2 was found to be the nucleotide sequence shown in SEQ ID NO:158 of the sequence listing (Fig. 154).

The nucleotide sequence of p_C3E-7085-LC was reanalyzed and the nucleotide sequence of full-length C3E-7085-LC was found to be the nucleotide sequence shown in SEQ ID NO:159 of the sequence listing (Fig. 155).

Based on the nucleotide sequences presented above, the amino acid sequences of the full-length NYZ-1010-HC2 and C3E-7085-LC encoded by them are identified.

The amino acid sequence of the full-length NYZ-1010-HC2 is the amino acid sequence shown in SEQ ID NO:160 of the sequence listing (Fig. 156). In such an amino acid sequence, amino acids 1-19 constitute a signal sequence, amino acids 21-271 constitute NYA-3061, amino acids 272-276 constitute a linker, and amino acids 277-394 constitute the heavy chain variable region C3E-7085. In addition, amino acids 395-724 constitute a constant region.

The amino acid sequence of the full-length C3E-7085-LC is the amino acid sequence shown in SEQ ID NO: 161 of the sequence listing (Fig. 157). In such amino acid sequence, amino acids 1-20 constitute a signal sequence, amino acids 21-127 constitute a variable region, and amino acids 128-233 constitute a constant region. In addition, amino acids 46-53 constitute CDRL1 (SEQ ID NO: 144 (Fig. 142)), amino acids 71-73 constitute CDRL2 (SEQ ID NO: 145 (Fig. 142)), and amino acids 110-117 constitute CDRL3 (SEQ ID NO: 146 (Fig. 142)).

12)-2 Expression and purification of various Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecules by mutagenesis and format modification or NYF-0061

In the same manner as in 7)-2-1, the culture supernatant of the anti-HLA/NY-ESO-anti-CD3 taFv-Fc-type heterodimer bispecific molecule (NYZ-0038) was prepared using p_NYZ-0038-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYZ-0038 are shown in SEQ ID NO: 155 of the sequence listing (Fig. 151) and SEQ ID NO: 84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc type heterodimer molecule (NYZ-0082) was prepared using p_NYZ-0082-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYZ-0082 are shown in SEQ ID NO:156 of the sequence listing (Fig. 152) and SEQ ID NO:84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 taFv-Fc-type heterodimer molecule (NYZ-0083) was prepared using p_NYZ-0083-HC2 and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYZ-0083 are shown in SEQ ID NO: 157 of the sequence listing (Fig. 153) and in SEQ ID NO: 84 of the sequence listing (Fig. 85).

The culture supernatant of the bispecific anti-HLA/NY-ESO-anti-CD3 scFv-Fab-Fc type heterodimer molecule (NYZ-1010) was prepared using p_NYZ-1010-HC2, p_C3E-7085-LC and p_HC1. The amino acid sequences obtained by the expression vectors constituting NYZ-1010 are shown in SEQ ID NO: 160 of the sequence listing (Fig. 156), SEQ ID NO: 161 of the sequence listing (Fig. 157) and SEQ ID NO: 84 of the sequence listing (Fig. 85).

The culture supernatants obtained above are purified in the same manner as in 7)-3. The purified protein samples are subjected to analytical SEC, the degree of purification and concentration are determined, and then the samples are subjected to various types of evaluation.

(Example 13) Evaluation of HLA/NY-ESO binding affinity using Biacore

Using Biacore T200, various Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecules were captured as ligands to the immobilized anti-human IgG antibody (Fc), and the antigen was analyzed as an analyte. HLA/NY-ESO prepared in 1)-1 was used as the antigen. Anti-human IgG (Fc) antibody (Human Antibody Capture Kit, GE Healthcare) was immobilized on the CM5 sensor chip (GE Healthcare) according to the kit instructions. Fc-conjugated anti-HLA/NY-ESO-anti-CD3 bispecific molecules diluted to 0.5 μg/mL in HBS-EP+ (GE Healthcare) for evaluation were contacted therewith at 10 μL/min for 60 seconds for immobilization. The samples were then added to HLA/NY-ESO analytes diluted to various levels with HBS-EP+ at a flow rate of 30 μL/min for 120 seconds and analyzed for dissociation for 600 seconds. The calculated results from this single cycle kinetics analysis, K _D , are shown in Table 7. NYZ-0038, NYZ-0082, NYZ-0083, and NYZ-1010 retained binding affinities equivalent to those of NYF-0061.

[Table 7]

Clone name K _D (nM) NYF-0061 1.5 NYZ-0038 1.5 NYZ-0082 1.8 NYZ-0083 1.7 NYZ-1010 1.9

(Example 14) Generation of CD3e-knockout human T2 lymphoblast fusion cells

To knock out CD3e in the genomic sequence of human T2 lymphoblast fusion cells (ATCC) by CRISPR, Cas9 expression plasmid (GE Healthcare) and enRNA expression plasmid (LONZA) were introduced by electroporation. The cells were then cloned by limiting dilution. Cells in which deletion of the target gene fragment from the introduced cells and CD3e gene expression were not observed by genomic analysis and RT-PCR analysis of total RNA, respectively, were subjected to subsequent experiments.

(Example 15) Amino acid recognition analysis of the NY-ESO Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule peptide

The CD3e knockout T2 cells obtained in Example 14 were adjusted to an adequate concentration in AIM-V medium (Thermo Fisher Scientific) containing 20% FBS, NY-ESO peptide (SEQ ID NO:1), NY-ESO point mutation peptides 1F, 2M, 3A, 4A, 5A, 6L, 7F, 8A and 9A (SEQ ID NOs:121 (Fig. 122), 122 (Fig. 123), 123 (Fig. 124), 124 (Fig. 125), 125 (Fig. 126), 126 (Fig. 127), 127 (Fig. 128), 128 (Fig. 129) and 129 (Fig. 130)) and peptide gp100 (SEQ ID NO:130 (Fig. 131)) (Sigma Genosys) was added in the same manner as in Example 5, cells stained with the LIVE/DEAD Fixable Dead Cell Stain Kit (Thermo Fisher Scientific) were divided into two groups, cells in PBS containing 5% FBS were seeded in a 96-well U-bottom microplate at ¹⁰⁵ cells/well, and the plate was centrifuged followed by removal of the supernatant. The various Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules prepared in Example 12 and diluted to 100 nM with PBS containing 5% FBS were added at 25 μl/well to one of the two groups, and the resulting product was incubated at 4°C for 30 minutes. The cells were washed twice in PBS containing 5% FBS, PE-conjugated F(ab')2 fragment against human IgG (Jackson Immuno Research Laboratories) diluted in PBS containing 5% FBS was added at 25 μl/well, and the resulting product was incubated at 4°C for 30 min. The cells were washed twice in PBS containing 5% FBS, immobilized with Mildform 10 N (FUJIFILM Wako Pure Chemical Corporation) overnight, and resuspended in PBS containing 5% FBS. For standardization of the amount of HLA/peptide complex, HLA-A2 antibody BB7.2-Alexa Fluor 488 diluted to 10 μg/ml in PBS containing 5% FBS or mouse IgG2b-Alexa Fluor 488 (BioRAD) was added at 25 μl/well and the mixture was incubated at 4°C for 30 min. The cells were washed 2 times in PBS containing 5% FBS, immobilized with Mildform 10 N (FUJIFILM Wako Pure Chemical Corporation) overnight and resuspended in PBS containing 5% FBS. The cell suspensions were detected using a flow cytometer (CantoII, Becton Dickinson). Data analysis was performed using Flowjo (Treestar) and the geometric mean fluorescence intensity (gMFI) of PE or Alexa Fluor 488 was measured in CD3e knockout T2 cells from which dead cells were removed. The standardized gMFI, which represents the binding affinity of each Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule standardized to the amount of HLA/peptide complex on CD3e knockout T2 cells, was determined according to the following equation.

Standardized gMFI=(A/B)×((C/D)/(E/F))

A/B=relative gMFI

A: gMFI PE in CD3e knockout T2 cells supplemented with DMSO or peptide in the presence of antibody

B: gMFI PE in CD3e knockout T2 cells supplemented with DMSO or peptide.

(C/D)/(E/F)=corrected amount of HLA/peptide complex in CD3e knockout T2 cells supplemented with DMSO or peptide.

C: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO or peptide in the presence of anti-HLA-A2 antibody.

D: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO or peptide in the presence of mouse IgG2b antibody.

E: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO containing HLA-A2 antibody.

F: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO containing mouse IgG2b antibody.

As shown in Fig. 158A, Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0038, NYZ-0082, NYZ-0083, and NYZ-1010, exhibited binding affinity for CD3e knockout T2 cells complemented with peptides containing point mutations introduced at amino acids 1, 4, 5, and 7 that were reduced to a quarter of the binding affinity for the wild-type NY-ESO peptide or less. This indicates that such constructs recognize amino acids 1, 4, 5, and 7 of the NY-ESO peptide well.

(Example 16) Evaluation of the antigen-binding specificity of Fc-conjugated bispecific anti-HLA-A2/NY-ESO-anti-CD3 molecules

In the same manner as in Example 6), the 9-mer peptide shown in Fig. 2A exhibiting sequence identity with the peptide NY-ESO:SLLMWITQC (SEQ ID NO:1) at amino acids 1, 4, and 5, which were shown to be well recognized by four types of antibodies, was selected as a homologous peptide and the binding specificity thereto was evaluated. CD3e knockout T2 cells were adjusted to an adequate concentration in AIM-V medium (Thermo Fisher Scientific) containing 20% FBS, NY-ESO peptide solution (SEQ ID NO:1), DOLPP1, IL20RB, PRKD2, CD163, and P2RY8 homologous peptides (SEQ ID NO:131 (Fig. 132), SEQ ID NO:132 (Fig. 133), SEQ ID NO:133 (Fig. 134), SEQ ID NO:134 (Fig. 135), and SEQ ID NO:135 (Fig. 136)), or gp100 peptide (SEQ ID NO:130 (Fig. 131)) (Sigma Genosys) dissolved to 5 mM in DMSO was added to give a final concentration of 50 μM or DMSO at 1/100, and the binding affinity of the antibodies was estimated in the same manner as in Example 15. The standardized gMFI indicating the binding affinity of each Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecule standardized for the amount of HLA/peptide complex on CD3e knockout T2 cells was determined according to the following equation.

Standardized gMFI=(A/B)×((C/D)/(E/F))

A: gMFI PE in CD3e knockout T2 cells supplemented with DMSO or peptide in the presence of antibody

B: gMFI PE in CD3e knockout T2 cells supplemented with DMSO or peptide.

(C/D)/(E/F)=corrected amount of HLA/peptide complex in CD3e knockout T2 cells supplemented with DMSO or peptide.

C: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO or peptide in the presence of HLA-A2 antibody.

D: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO or mouse IgG2b antibody-containing peptide.

E: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO containing HLA-A2 antibody.

F: gMFI Alexa488 in CD3e knockout T2 cells supplemented with DMSO containing mouse IgG2b antibody.

As shown in Fig. 158B, none of the Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules; i.e., NYZ-0038, NYZ-0082, NYZ-0083, and NYZ-1010, bind to cells complemented with homologous peptides, and these molecules exhibit very high specificity.

(Example 17) Evaluation of the cytotoxicity of Fc-conjugated bispecific anti-HLA-A2/NY-ESO-anti-CD3 molecules

17)-1 Obtaining target cells

In the same manner as in Example 8)-1, suspensions of endogenous cells expressing human NY-ESO (U266B1 and NCI-H1703) and cells not expressing endogenous human NY-ESO (AGS and CFPAC-1) were prepared and used as target cells.

17)-2 Obtaining effector cells

In the same manner as in Example 8)-2, a suspension of commercially available frozen human PBMCs (Cellular Technology Limited) was prepared and used as effector cells.

17)-3 Cytotoxicity assay

The target cells obtained in Example 17)-1 were seeded in a 96-well U-bottom microplate at 50 μl/well. Various Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules obtained in Example 12 and adjusted to different concentration levels were added thereto at 50 μl/well, the effector cells obtained in Example 17)-2 were added thereto at 100 μl/well, centrifuged at room temperature and 1000 rpm for 1 minute, and then incubated at 37°C in the presence of 5% CO.₂for 20-24 hours. The supernatant (50 µl) is collected on a LumaPlate (PerkinElmer), dried at 50°C for approximately 2 hours, and then analyzed using a plate reader (TopCount, PerkinElmer). The test is performed in triplicate, and the rate of cell lysis is determined according to the following equation.

Cell lysis rate (%) = (A - B) / (C - B) × 100

A: number of sample wells

B: Average background value (wells without added antibodies) (n=3).

When the antibody is added, 50 µl of assay medium is added. The remaining procedures are the same as for the sample wells.

C: mean maximum release value (wells in which target cells are lysed by surfactant) (n=3).

When the antibody is added, 50 µl of assay medium is added. Surfactant is added at 100 µl, a 50 µl fraction is transferred to the LumaPlate as for the sample wells, and assays are performed.

As shown in Fig. 159A to Fig. 159D, different anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules exhibited cytotoxicity against endogenous cells expressing human NY-ESO (U266B1 and NCI-H1703). The EC ₅₀ calculation results obtained using the four-parameter logistic curve equation of the analytical software (Sigmaplot, version 12.0) for U266B1 cells are shown in Table 8, and the results for NCI-H1703 cells are shown in Table 9. As shown in Fig. 159E to Fig. 159H, in contrast, no cytotoxicity was observed on endogenous cells not expressing human NY-ESO (AGS and CFPAC-1).

[Table 8]

Name EC ₅₀ (nM) 1 NYZ-0038 1.01 2 NYZ-0082 0.45 3 NYZ-0083 0.25 4 NYZ-1010 0.96

[Table 9]

Name EC ₅₀ (nM) 1 NYZ-0038 0.61 2 NYZ-0082 1.00 3 NYZ-0083 0.71 4 NYZ-1010 2.56

(Example 18) In vivo evaluation of the activity of Fc-conjugated anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules in PBMC-transfected human models

Human squamous cell lung cancer cell lines NCI-H1703 (ATCC) were adjusted to 6× ¹⁰⁷ cells/mL in PBS containing 50% Matrigel (Corning), and 0.1 mL of this solution was injected subcutaneously into NOG mice (female, 6-7 weeks old) (day 0). On day 4, human PBMCs were adjusted to 3.75× ¹⁰⁷ -5× ¹⁰⁷ cells/mL in PBS, and 0.2 mL of this compound was injected intravenously. Approximately 1 week thereafter (day 6 - day 7), the measurement of the major diameter (mm) and minor diameter (mm) of the tumor was started using an electronic digital caliper over time, and the estimated tumor volume was calculated according to the equation below.

Estimated tumor volume ( ^mm3 ) = mean estimated tumor volume among individuals.

Estimated tumor volume in humans = major diameter x [minor diameter] ² /2

On day 14, the mice were divided into groups of 5 or 6 mice each depending on the tumor volume, and various anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules (NYZ-1010 at 1 mg/kg, NYZ-1010 at 1.2 mg/kg for comparison with the same molecular weight) were administered intravenously. The administration was performed on days 14 and 21. The antitumor effects were observed in the treatment groups administered with different anti-HLA-A2/NY-ESO-anti-CD3 bispecific molecules (Fig. 160A to Fig. 160D). The tumor growth inhibition (%) from day 28 to day 29 was calculated according to the following equation and is shown in Table 10.

Tumor growth inhibition (%) = 100 - (estimated tumor volume in the treatment group/estimated tumor volume in the control group × 100)

[Table 10]

Name Tumor growth inhibition (%) 1 NYZ-0038 100,0 2 NYZ-0082 100,0 3 NYZ-0083 99.8 4 NYZ-1010 99.9

(Example 19) Comparison of the physicochemical properties of different Fc-conjugated bispecific anti-HLA/NY-ESO-anti-CD3 molecules

19)-1 Evaluation of acid treatment

NYF-0016 and various CDR-grafted mutants of NYA-1143 (NYF-0044, NYF-0045, NYF-0047, NYF-0048, NYF-0060, and NYF-0061) obtained in 7)-2-1 were concentrated by centrifugation using Amicon (Millipore), the buffer was exchanged with 25 mM sodium acetate, 5% sorbitol (pH 5.5), and the concentration was adjusted to 15 mg/ml (NYF-0016 at 10 mg/ml). The samples were then dialyzed using an Xpress Micro Dialyzer (Scienova) against 100 mM sodium acetate (pH 3.5) or 25 mM sodium acetate, 5% sorbitol (pH 5.5) as a control, and the samples were collected and incubated at room temperature for 1 hour. The pH levels of the samples were then adjusted to 5.0 using 500 mM Tris-HCl (pH 9.0). The samples were analyzed by size exclusion chromatography (SEC) using an ACQUITY UPLC BEH200 SEC 1.7 μm 4.6*150 mm (Waters). The mobile phase was 0.2 M Ki/200 mM KCl/pH 7.0, and the analysis was performed at a flow rate of 0.2 ml/min (detection wavelength: 280 nm). The peak content (%) of polymers contained in the samples was analyzed and calculated by the area percentage method. The results are shown in Fig. 162. As a result of the acid treatment, the polymer content of NYF-0016 increased to 96%. In contrast, in various grafted mutants of NYA-1143 CDR, the amount of polymer formed by the acid treatment decreased. Specifically, in NYF-0047, NYF-0048, NYF-0060, and NYF-0061, the amount of polymer formed by the acid treatment decreased to 1% or less. This indicates a significant improvement in acid resistance.

19)-2 Evaluation of solution stability

NYF-0061 obtained in 7)-2-1 and NYZ-0038, NYZ-0082, NYZ-0083 and NYZ-1010 obtained in 12)-2 were concentrated by centrifugation using an Amicon Ultra-4 (Millipore), the buffer was exchanged with 25 mM histidine, 5% sorbitol (pH 6.0), 25 mM histidine and 5% sorbitol (pH 6.0) were added thereto, and the sample concentration was adjusted to 25 mg/mL to prepare evaluation samples. First, the high molecular weight compounds (HMWS) of each evaluation sample were determined by the area percentage method by size exclusion chromatography using an AdvanceBio SEC 300A 2.7 μm 4.6×300 mm (Agilent). 0.2 M Ki/200 mM KCl/pH 7.0 was used as the mobile phase, and the analysis was carried out at a flow rate of 0.2 ml/min (detection wavelength: 280 nm). The solution stability test of each sample was carried out by storing at 25°C for 6 days (NYF-0061: 7 days), size exclusion chromatography under the conditions described above, and calculating the HMWS (%) of each sample by the area percentage method. The results are shown in Table 11. The HMWS (%) of NYF-0061 increased to approximately 10.6% as the storage time progressed. In contrast, the increase in HMWS (%) of NYZ-0038, NYZ-0082, NYZ-0083, and NYZ-1010 was approximately 2% or less. This shows that the stability of NYZ-0038, NYZ-0082, NYZ-0083 and NYZ-1010 solutions is higher than that of NYF-0061.

[Table 11]

Clone name Navy (%) NYF-0061 10.6 NYZ-0038 2.1 NYZ-0082 1.7 NYZ-0083 1.9 NYZ-1010 0.8

(Example 20) Preparation of anti-HLA/NY-ESO scFv

20)-1 Construction of anti-HLA/NY-ESO scFv expression vector

scFv antibody constructs are designed that can have high binding affinity to HLA-A2/NY-ESO-1; i.e., mAb24955N, mAb24956N, mAb28075P, mAb28105P, mAb28113P and mAb29822P2 (WO2021/003357), and scFv mAb24955N is designated as NYC-0005, scFv mAb24956N is designated as NYC-0006, scFv mAb28075P is designated as NYC-0007, scFv mAb28105P is designated as NYC-0008, scFv mAb28113P is designated as NYC-0009 and scFv mAb29822P2 is designated as NYC-0010.

In addition, scFv expression vectors for NYC-0005, NYC-0006, NYC-0007, NYC-0008, NYC-0009, and NYC-0010, each containing pcDNA3.4 (ThermoFisher Scientific) as a backbone, were constructed in mammalian cells. In order to compare the expression levels of the same vector backbones and formats in mammalian cells, scFv expression vectors for NYA-2047, NYA-2061, and NYA-3061 were also constructed in mammalian cells.

The nucleotide sequences of the constructed scFv expression vectors were reanalyzed, and the nucleotide sequences of full-length NYA-2047, NYA-2061, NYA-3061, NYC-0005, NYC-0006, NYC-0007, NYC-0008, NYC-0009, and NYC-0010 were found to be the nucleotide sequences shown in SEQ ID NO:43 (Fig. 50), SEQ ID NO:46 (Fig. 53), SEQ ID NO:163 (Fig. 164), SEQ ID NO:165 (Fig. 166), SEQ ID NO:167 (Fig. 168), SEQ ID NO:169 (Fig. 170), SEQ ID NO:171 (Fig. 172), SEQ ID NO:173 (Fig. 174) and SEQ ID NO: 175 (Fig. 176) of the sequence listing. Based on the nucleotide sequences mentioned above, the amino acid sequences encoded by them are as follows: the amino acid sequence of the full-length NYA-2047 is shown in SEQ ID NO: 50 (Fig. 57); the amino acid sequence of the full-length NYA-2061 is shown in SEQ ID NO: 53 (Fig. 60); the amino acid sequence of the full-length NYA-3061 is shown in SEQ ID NO: 164 (Fig. 165); the amino acid sequence of the full-length NYC-0005 is shown in SEQ ID NO: 166 (Fig. 167); the amino acid sequence of the full-length NYC-0006 is shown in SEQ ID NO: 168 (Fig. 169); the amino acid sequence of full-length NYC-0007 is shown in SEQ ID NO:170 (FIG. 171); the amino acid sequence of full-length NYC-0008 is shown in SEQ ID NO:172 (FIG. 173); the amino acid sequence of full-length NYC-0009 is shown in SEQ ID NO:174 (FIG. 175); and the amino acid sequence of full-length NYC-0010 is shown in SEQ ID NO:176 (FIG. 177).

20)-2 Expression and purification of anti-HLA/NY-ESO scFv

The gene culture and transfection in Expi293F cells (ThermoFisher Scientific) were performed in the same manner as described in 1)-5-2. To estimate the amount of production, the cells were subjected to shaking culture in an incubator at 37°C in the presence of 8% _CO2 at 135 rpm for 4 days after gene introduction, the culture supernatant was filtered through a 0.2 μm filter (ThermoFisher Scientific), and the scFv anti-HLA/NY-ESO culture supernatant was obtained. Purification was performed using Ni Sepharose excel (Cytiva), the eluate was subjected to analytical size exclusion chromatography (SEC), and the concentration was determined. The production amounts are shown in Table 12. The production amount of NYA-2047, NYA-2061 and NYA-3061 determined using Ni Sepharose excel relative to 1 L of culture supernatant in the cleared eluate was higher than that of NYC-0005, NYC-0006, NYC-0007, NYC-0008, NYC-0009 and NYC-0010.

[Table 12]

Clone name Amount produced per 1 liter (mg) NYA-2047 53.3 NYA-2061 45.7 NYA-3061 52.5 NYC-0005 2.5 NYC-0006 1,1 NYC-0007 10.8 NYC-0008 0.5 NYC-0009 2.4 NYC-0010 1.8

The eluate of each scFv obtained using Ni Sepharose excel was concentrated and purified through a gel filtration column (Superdex 200 Increase, Cytiva) equilibrated with 25 mM histidine, 300 mM NaCl, 5% sorbitan (pH 5.5). The purified protein samples were subjected to analytical SEC, the degree of purification and concentration were determined, and then the samples were subjected to various types of evaluation.

(Example 21) Expression and purification of scFv-heterodimer-Fc anti-HLA/NY-ESO

21)-1 Construction of scFv-heterodimer-Fc expression vector anti-HLA/NY-ESO

To compare the productivity obtained from different formats, an expression vector was constructed by converting the anti-HLA/NY-ESO scFv format into an Fc fusion type. The heterodimeric Fc sequences (hereafter referred to as HC-h and HC-k) were those reported in Nat. Biotechnol., Jul 1998, 16 (7), 677-81.

ScFv-heterodimer-Fc antibody constructs are being designed that may have high binding affinity to HLA-A2/NY-ESO-1; i.e. mAb24955N, mAb24956N, mAb28075P, mAb28105P, mAb28113P and mAb29822P2 (WO 2021/003357), and scFv-heterodimer-Fc mAb24955N are designated as NYC-0011, scFv-heterodimer-Fc mAb24956N is designated as NYC-0012, scFv-heterodimer-Fc mAb28075P is designated as NYC-0013, scFv-heterodimer-Fc mAb28105P is designated as NYC-0014, scFv-heterodimer-Fc mAb28113P is designated as NYC-0015 and scFv-heterodimer-Fc mAb29822P2 is designated as NYC-0016. Also, scFv-heterodimer-Fc NYA-2047 is designated as NYD-2047, scFv-heterodimer-Fc NYA-2061 is designated as NYD-2061, and scFv-heterodimer-Fc NYA-3061 is designated as NYD-3061.

Expression vectors for HC-h, NYC-0011-HC-k, NYC-0012-HC-k, NYC-0013-HC-k, NYC-0014-HC-k, NYC-0015-HC-k, and NYC-0016-HC-k, each containing pcDNA3.3 or pcDNA3.4 (ThermoFisher Scientific) as a mammalian cell backbone were constructed. Additionally, mammalian cell expression vectors NYD-2047-HC-k, NYD-2061-HC-k, and NYD-3061-HC-k were constructed to compare expression levels in mammalian cells with the same vector backbones and formats.

The nucleotide sequences of the constructed scFv-heterodimer-Fc expression vectors, and the nucleotide sequences of full-length HC-h, NYD-2047-HC-k, NYD-2061-HC-k, NYD-3061-HC-k, NYC-0011-HC-k, NYC-0012-HC-k, NYC-0013-HC-k, NYC-0014-HC-k, NYC-0015-HC-k and NYC-0016-HC-k are the nucleotide sequences shown in SEQ ID NO: 177 (FIG. 178), SEQ ID NO: 179 (FIG. 180), SEQ ID NO: 181 (FIG. 182), SEQ ID NO: 183 (FIG. 184), SEQ ID NO: 185 (Fig. 186), SEQ ID NO: 187 (Fig. 188), SEQ ID NO: 189 (Fig. 190), SEQ ID NO: 191 (Fig. 192), SEQ ID NO: 193 (Fig. 194) and SEQ ID NO: 195 (Fig. 196) of the sequence listing. Based on the nucleotide sequences mentioned above, the full-length amino acid sequences encoded by them are as follows: the amino acid sequence of the whole HC-h is shown in SEQ ID NO: 178 (Fig. 179); the amino acid sequence of the full-length NYD-2047-HC-k is shown in SEQ ID NO: 180 (Fig. 181); the amino acid sequence of the full-length NYD-2061-HC-k is shown in SEQ ID NO: 182 (Fig. 183); the amino acid sequence of the full-length NYD-3061-HC-k is shown in SEQ ID NO: 184 (Fig. 185); the amino acid sequence of the full-length NYC-0011-HC-k is shown in SEQ ID NO: 186 (Fig. 187); the amino acid sequence of the full-length NYC-0012-HC-k is shown in SEQ ID NO: 188 (Fig. 189); the amino acid sequence of the full-length NYC-0013-HC-k is shown in SEQ ID NO: 190 (Fig. 191); the amino acid sequence of the full-length NYC-0014-HC-k is shown in SEQ ID NO: 192 (Fig. 193); the amino acid sequence of the full-length NYC-0015-HC-k is shown in SEQ ID NO: 194 (Fig. 195); and the amino acid sequence of full-length NYC-0016-HC-k is shown in SEQ ID NO:196 (Fig. 197).

21)-2 Expression and purification of scFv-heterodimer-Fc anti-HLA/NY-ESO

Cultivation and gene introduction into Expi293F cells (ThermoFisher Scientific) were carried out in the same manner as described in 1)-5-2. The plasmids used to obtain the clones are shown in Table 13 below.

[Table 13]

Clone name HC-k HC-h NYD-2047 NYD-2047-HC-k HC-h NYD-2061 NYD-2061-HC-k HC-h NYD-3061 NYD-3061-HC-k HC-h NYC-0011 NYC-0011-HC-k HC-h NYC-0012 NYC-0012-HC-k HC-h NYC-0013 NYC-0013-HC-k HC-h NYC-0014 NYC-0014-HC-k HC-h NYC-0015 NYC-0015-HC-k HC-h NYC-0016 NYC-0016-HC-k HC-h

To assess the amount of production, the cells were subjected to shaking culture incubator at 37°C in the presence of 8% _CO2 at 135 rpm for 4 days after gene introduction, the culture supernatant was filtered through 0.2 μm filter (ThermoFisher Scientific) to obtain scFv-heterodimer-Fc anti-HLA/NY-ESO culture supernatant. For purification, the culture supernatant was applied to MabSelectSuRe resin (Cytiva) equilibrated with PBS (pH 7.4) to allow scFv-heterodimer-Fc target anti-HLA/NY-ESO to be adsorbed thereon. After the non-adsorbed components were removed with PBS, the adsorbed components were eluted with acetate buffer, the eluate was neutralized with Tris buffer, and the eluate was subjected to analytical size exclusion chromatography (SEC) to determine the purity and concentration. Table 14 shows the production amount relative to 1 liter of culture supernatant. The production amount of NYA-2047, NYA-2061, and NYA-3061 determined using MabSelectSuRe resin per 1 liter of culture supernatant in the purified eluate was higher than that of NYC-0011, NYC-0012, NYC-0013, NYC-0014, NYC-0015, and NYC-0016.

[Table 14]

Clone name Amount produced per 1 liter (mg) NYD-2047 93.3 NYD-2061 112.2 NYD-3061 127.5 NYC-0011 17.2 NYC-0012 4.2 NYC-0013 54.1 NYC-0014 1.8 NYC-0015 12.2 NYC-0016 16.6

The eluate of each scFv-heterodimer-Fc produced using MabSelectSuRe resin was concentrated and purified on a gel filtration column (Superdex 200 Increase, Cytiva) equilibrated with 25 mM histidine, 300 mM NaCl, 5% sorbitan (pH 5.5). Purified protein samples were subjected to analytical size exclusion chromatography (SEC) and NYD-2047, NYD-2061, NYD-3061, NYC-0011, NYC-0013 and NYC-0015 were used as samples with sufficient purity and quantities to assess solution stability. Target was not detected in NYC-0016 within the elution time determined by SEC. Thus, it was determined that the target was not expressed in the culture supernatant.

(Example 22) Evaluation of the stability of scFv-heterodimer-Fc anti-HLA/NY-ESO solution

NYD-2047, NYD-2061, NYD-3061, NYC-0011, NYC-0013, and NYC-0015 obtained in Example 21 were concentrated by centrifugation using an Amicon Ultra-4 (Millipore), the buffer was replaced with PBS, PBS was added thereto, and the sample concentration was adjusted to 5 mg/mL to prepare samples for evaluation. First, the HMWS (%) of each sample to be evaluated was determined by the area percentage method by size exclusion chromatography using an AdvanceBio SEC 300A 2.7 μm 4.6×150 mm (Agilent). 0.2 M Ki/200 mM KCl/pH 7.0 was used as the mobile phase, and the analysis was performed at a flow rate of 0.2 mL/min (detection wavelength: 280 nm). An accelerated aging test was conducted on each sample by storing at 40°C for 1 day and 7 days, size exclusion chromatographing under the conditions described above, and calculating the HMWS (%) of each sample by the area percentage method. Fig. 163 shows the results showing the changes that occurred in each sample upon storage at 40°C over time.

The HMWS (%) of NYD-2047, NYD-2061 and NYD-3061 increases to 8.7%, 8.1% and 4.4%, respectively, after the storage time. In contrast, the HMWS (%) of NYC-0011, NYC-0013 and NYC-0015 increases significantly; 71.8%, 42.4% and 97.3%, respectively, after the storage time.

The results show that NYD-2047, NYD-2061 and NYD-3061 are superior to NYC-0011, NYC-0013 and NYC-0015 in terms of solution stability. In addition, the HMWS (%) of NYD-3061 is the lowest among the evaluated samples. That is, NYD-3061 is superior in terms of solution stability.

(Example 23) Evaluation of HLA/NY-ESO binding affinity using Biacore

NYC-0005, NYC-0007 and NYC-0008, equivalent to the scFvs comprising the anti-HLA/NY-ESO NYC-0011, NYC-0013 and NYC-0015, were used to assess binding affinity to HLA/NY-ESO.

Using Biacore T200, anti-HLA/NY-ESO scFv was captured as a ligand to immobilized anti-His antibody, and the antigen was analyzed as an analyte. HLA/NY-ESO prepared in 1)-1 was used as the antigen. Anti-His antibody (His Capture Kit, Cytiva) was immobilized on the CM5 sensor chip (Cytiva) according to the kit instructions. Anti-HLA/NY-ESO scFv constructs to be tested diluted to 0.5 μg/mL in HBS-EP+ (Cytiva) were contacted therewith at 10 μL/min for 60 seconds for immobilization. Thereafter, the samples were added to HLA/NY-ESO analytes diluted to different levels with HBS-EP+ at a flow rate of 30 μL/min for 120 seconds and the dissociation was analyzed for 600 seconds. The calculated results obtained from this single-cycle kinetics analysis, K _D , are shown in Table 15. It was confirmed that NYC-0005, NYC-0007, and NYC-0008 would bind to HLA/NY-ESO under the conditions in which the binding affinity of NYA-2047, used as a positive control, to HLA/NY-ESO was consistently confirmed.

[Table 15]

Clone name To _D NYC-0005 9.3 nM NYC-0007 1.0 µM NYC-0008 7.9 nM

Industrial applicability

The bispecific antibody of the present invention can be used as a therapeutic or prophylactic agent against cancer and the like.

All publications, patents and patent applications cited in this document are hereby incorporated by reference in their entirety.

Free text from sequence list

SEQ ID NO 1: amino acid sequence of the peptide in NY-ESO (Fig. 8).

SEQ ID NO 2: amino acid sequence of the peptide in MAGEC-1 (Fig. 9).

SEQ ID NO 3: Primer 1 for scFv sequence analysis (Fig. 10).

SEQ ID NO 4: Primer 2 for scFv sequence analysis (Fig. 11).

SEQ ID NO 5: nucleotide sequence of the variable region of the heavy chain of NYA-0001 (Fig. 12).

SEQ ID NO 6: amino acid sequence of the variable region of the heavy chain of NYA-0001 (Fig. 13).

SEQ ID NO 7: nucleotide sequence of the variable region of the light chain of NYA-0001 (Fig. 14).

SEQ ID NO:8: amino acid sequence of the light chain variable region of NYA-0001 (Fig. 15).

SEQ ID NO 9: nucleotide sequence of the variable region of the heavy chain of NYA-0060 (Fig. 16).

SEQ ID NO 10: amino acid sequence of the variable region of the heavy chain of NYA-0060 (Fig. 17).

SEQ ID NO 11: nucleotide sequence of the light chain variable region of NYA-0060 (Fig. 18).

SEQ ID NO 12: amino acid sequence of the light chain variable region of NYA-0060 (Fig. 19).

SEQ ID NO 13: nucleotide sequence of the variable region of the heavy chain of NYA-0068 (Fig. 20).

SEQ ID NO 14: amino acid sequence of the variable region of the heavy chain of NYA-0068 (Fig. 21).

SEQ ID NO 15: nucleotide sequence of the variable region of the light chain of NYA-0068 (Fig. 22).

SEQ ID NO 16: amino acid sequence of the light chain variable region of NYA-0068 (Fig. 23).

SEQ ID NO 17: nucleotide sequence of the variable region of the heavy chain of NYA-0082 (Fig. 24).

SEQ ID NO 18: amino acid sequence of the variable region of the heavy chain of NYA-0082 (Fig. 25).

SEQ ID NO 19: nucleotide sequence of the light chain variable region of NYA-0082 (Fig. 26).

SEQ ID NO 20: amino acid sequence of the light chain variable region of NYA-0082 (Fig. 27).

SEQ ID NO 21: Nucleotide sequence of the NYA-1163 tag adduct (Fig. 28).

SEQ ID NO 22: Nucleotide sequence of the NYA-2023 tag adduct (Fig. 29).

SEQ ID NO 23: Nucleotide sequence of the NYA-2027 tag adduct (Fig. 30).

SEQ ID NO 24: Nucleotide sequence of the NYA-1143 tag adduct (Fig. 31).

SEQ ID NO 25: Nucleotide sequence of the NYA-2143 tag adduct (Fig. 32).

SEQ ID NO 26: amino acid sequence of the NYA-1163 tag adduct (Fig. 33); NYA-1163: amino acids 21-266.

SEQ ID NO 27: amino acid sequence of the NYA-2023 tag adduct (Fig. 34); NYA-2023: amino acids 21-266.

SEQ ID NO 28: amino acid sequence of the NYA-2027 tag adduct (Fig. 35); NYA-2027: amino acids 21-266.

SEQ ID NO 29: amino acid sequence of the NYA-1143 tag adduct (Fig. 36); NYA-1143: amino acids 21-266.

SEQ ID NO 30: amino acid sequence of the NYA-2143 tag adduct (Fig. 37); NYA-2143: amino acids 21-266.

SEQ ID NO 31: Nucleotide sequence of the NYA-1154 tag adduct (Fig. 38).

SEQ ID NO 32: amino acid sequence of the NYA-1154 tag adduct (Fig. 39); NYA-1154: amino acids 21-266.

SEQ ID NO 33: truncated amino acid sequence of HLA-A*0201 (GenBank: ASA47534.1) (Fig. 40).

SEQ ID NO 34: amino acid sequence of β2-microglobin (Fig. 41).

SEQ ID NO 35: Nucleotide sequence of the NYA-2035 tag adduct (Fig. 42).

SEQ ID NO 36: amino acid sequence of the NYA-2035 tag adduct (Fig. 43); NYA-2035: amino acids 21-266.

SEQ ID NO 37: amino acid sequence of NYA-1143-VH01 (Fig. 44).

SEQ ID NO 38: amino acid sequence of NYA-1143-VH02 (Fig. 45).

SEQ ID NO 39: amino acid sequence of NYA-1143-VH03 (Fig. 46).

SEQ ID NO 40: amino acid sequence of NYA-1143-VL01 (Fig. 47).

SEQ ID NO 41: Nucleotide sequence of the NYA-2044 tag adduct (Fig. 48).

SEQ ID NO 42: nucleotide sequence of the NYA-2045 tag adduct (Fig. 49).

SEQ ID NO 43: Nucleotide sequence of the NYA-2047 tag adduct (Fig. 50).

SEQ ID NO 44: nucleotide sequence of the NYA-2048 tag adduct (Fig. 51).

SEQ ID NO 45: Nucleotide sequence of the NYA-2060 tag adduct (Fig. 52).

SEQ ID NO 46: nucleotide sequence of the NYA-2061 tag adduct (Fig. 53).

SEQ ID NO 47: amino acid sequence of the NYA-2044 tag adduct (Fig. 54); NYA-2044: amino acids 21-266.

SEQ ID NO 48: amino acid sequence of the NYA-2045 tag adduct (Fig. 55); NYA-2045: amino acids 21-266.

SEQ ID NO 49: amino acid sequence of NYA-0082 (Fig. 56).

SEQ ID NO 50: amino acid sequence of the NYA-2047 tag adduct (Fig. 57); NYA-2047: amino acids 21-266.

SEQ ID NO 51: amino acid sequence of the NYA-2048 tag adduct (Fig. 58); NYA-2048: amino acids 21-266.

SEQ ID NO 52: amino acid sequence of the NYA-2060 tag adduct (Fig. 59); NYA-2060: amino acids 21-266.

SEQ ID NO 53: amino acid sequence of the NYA-2061 tag adduct (Fig. 60); NYA-2061: amino acids 21-266.

SEQ ID NO 54: CDRH1 of heavy chain NYA-0001 (Fig. 61).

SEQ ID NO 55: CDRH2 of heavy chain NYA-0001 (Fig. 61).

SEQ ID NO 56: CDRH3 of heavy chain NYA-0001 (Fig. 61).

SEQ ID NO 57: CDRL1 light chain NYA-0001 (Fig. 61).

SEQ ID NO 58: CDRL2 light chain NYA-0001 (Fig. 61).

SEQ ID NO 59: CDRL3 light chain NYA-0001 (Fig. 61).

SEQ ID NO 60: amino acid sequence of CDRL1 NYA-2023 (Fig. 62).

SEQ ID NO 61: amino acid sequence of CDRL3 NYA-2027 (Fig. 63).

SEQ ID NO 62: CDRH3 of NYA-1154 heavy chain (Fig. 64).

SEQ ID NO 63: CDRL3 light chain of NYA-1154 (Fig. 64).

SEQ ID NO 64: amino acid sequence of CDRL1 NYA-0035 (Fig. 65).

SEQ ID NO 65: nucleotide sequence of the NYC-0003 tag adduct (Fig. 66).

SEQ ID NO 66: nucleotide sequence of the NYC-0004 tag adduct (Fig. 67).

SEQ ID NO 67: amino acid sequence of the adduct of the NYC-0003 tag (Fig. 68); NYC-0003: amino acids 21-263.

SEQ ID NO 68: amino acid sequence of the adduct of the NYC-0004 tag (Fig. 69); NYC-0004: amino acids 21-263.

SEQ ID NO 69: nucleotide sequence of the NYA-0001 tag adduct (Fig. 70).

SEQ ID NO 70: amino acid sequence of the adduct of the NYA-0001 tag (Fig. 71); NYA-0001: amino acids 21-266.

SEQ ID NO 71: nucleotide sequence of HC1 (Fig. 72).

SEQ ID NO 72: nucleotide sequence of NYF-0016-HC2 (Fig. 73).

SEQ ID NO 73: nucleotide sequence of NYF-0019-HC2 (Fig. 74).

SEQ ID NO 74: nucleotide sequence of NYF-0022-HC2 (Fig. 75).

SEQ ID NO 75: nucleotide sequence of NYF-0023-HC2 (Fig. 76).

SEQ ID NO 76: nucleotide sequence of NYF-0027-HC2 (Fig. 77).

SEQ ID NO 77: nucleotide sequence of NYF-0035-HC2 (Fig. 78).

SEQ ID NO 78: nucleotide sequence of NYF-0044-HC2 (Fig. 79).

SEQ ID NO 79: nucleotide sequence of NYF-0045-HC2 (Fig. 80).

SEQ ID NO 80: nucleotide sequence of NYF-0047-HC2 (Fig. 81).

SEQ ID NO 81: nucleotide sequence of NYF-0048-HC2 (Fig. 82).

SEQ ID NO 82: nucleotide sequence of NYF-0060-HC2 (Fig. 83).

SEQ ID NO 83: nucleotide sequence of NYF-0061-HC2 (Fig. 84).

SEQ ID NO 84: amino acid sequence of HC1 (Fig. 85).

SEQ ID NO 85: amino acid sequence of NYF-0016-HC2 (Fig. 86); NYA-1143: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 86: amino acid sequence of NYF-0019-HC2 (Fig. 87); NYA-2143: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 87: amino acid sequence of NYF-0022-HC2 (Fig. 88); NYA-1163: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 88: amino acid sequence of NYF-0023-HC2 (Fig. 89); NYA-2023: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 89: amino acid sequence of NYF-0027-HC2 (Fig. 90); NYA-2027: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 90: amino acid sequence of NYF-0035-HC2 (Fig. 91); NYA-2035: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 91: amino acid sequence of NYF-0044-HC2 (Fig. 92); NYA-2044: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 92: amino acid sequence of NYF-0045-HC2 (Fig. 93); NYA-2045: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 93: amino acid sequence of NYF-0047-HC2 (Fig. 94); NYA-2047: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 94: amino acid sequence of NYF-0048-HC2 (Fig. 95); NYA-2048: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 95: amino acid sequence of NYF-0060-HC2 (Fig. 96); NYA-2060: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 96: amino acid sequence of NYF-0061-HC2 (Fig. 97); NYA-2061: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 97: nucleotide sequence of the deletion NYA-0001-Fab-HC1-k (Fig. 98).

SEQ ID NO 98: nucleotide sequence of NYA-0001-LC (Fig. 99).

SEQ ID NO 99: amino acid sequence of deletion NYA-0001-Fab-HC1-k (Fig. 100); variable region of the heavy chain of NYA-0001: amino acids 20-139

SEQ ID NO 100: amino acid sequence of NYA-0001-LC (Fig. 101); variable region of the light chain of NYA-0001: amino acids 21-131

SEQ ID NO 101: nucleotide sequence of the deletion of NYA-1143-Fab-HC1-k (Fig. 102).

SEQ ID NO 102: nucleotide sequence of NYA-1143-LC (Fig. 103).

SEQ ID NO 103: nucleotide sequence of C3E-7085-HC2-k with deletion C (Fig. 104).

SEQ ID NO 104: amino acid sequence of NYA-1143-Fab-HC1-k deletion (Fig. 105); NYA-1143 heavy chain variable region: amino acids 20-139

SEQ ID NO 105: amino acid sequence of NYA-1143-LC (Fig. 106); variable region of the light chain of NYA-1143: amino acids 21-131

SEQ ID NO 106: amino acid sequence of the deletion C3E-7085-HC2-k (Fig. 107); C3E-7085: amino acids 21-260.

SEQ ID NO 107: nucleotide sequence of the NYA-1143-HC1-k deletion (Fig. 108).

SEQ ID NO 108: amino acid sequence of the deletion NYA-1143-HC1-k (Fig. 109); NYA-1143: amino acids 21-266.

SEQ ID NO 109: nucleotide sequence of deletion C3E-7085-NYA-1154-Fab-HC2-k (Fig. 110).

SEQ ID NO 110: nucleotide sequence of NYA-1154-LC (Fig. 111).

SEQ ID NO 111: nucleotide sequence of OAA-HC1-k deletion (Fig. 112).

SEQ ID NO 112: amino acid sequence of the deletion C3E-7085-NYA-1154-Fab-HC2-k (Fig. 113); C3E-7085: amino acids 21-260; heavy chain variable region NYA-1154: amino acids 266-285

SEQ ID NO 113: amino acid sequence of NYA-1154-LC (Fig. 114); variable region of the light chain of NYA-1154: amino acids 21-131

SEQ ID NO 114: amino acid sequence of OAA-HC1-k deletion (Fig. 115).

SEQ ID NO 115: nucleotide sequence of deletion NYF-0010-HC2-k (Fig. 116).

SEQ ID NO 116: nucleotide sequence of deletion NYF-0004-HC2-k (Fig. 117).

SEQ ID NO 117: nucleotide sequence of NYF-0011-HC2-k deletion (Fig. 118).

SEQ ID NO 118: amino acid sequence of the NYF-0010-HC2-k deletion (Fig. 119); NYA-1154: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 119: amino acid sequence of NYF-0004-HC2-k deletion (Fig. 120); C3E-7085: amino acids 21-260; and NYA-1154: amino acids 272-511.

SEQ ID NO 120: amino acid sequence of the NYF-0011-HC2-k deletion (Fig. 121); NYA-1143: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 121: amino acid sequence of the point mutant NY-ESO peptide 1F (Fig. 122).

SEQ ID NO 122: amino acid sequence of the point mutant peptide NY-ESO 2M (Fig. 123).

SEQ ID NO 123: amino acid sequence of the point mutant peptide 3A NY-ESO (Fig. 124).

SEQ ID NO 124: amino acid sequence of point mutant peptide 4A NY-ESO (Fig. 125).

SEQ ID NO 125: amino acid sequence of point mutant peptide 5A NY-ESO (Fig. 126).

SEQ ID NO 126: amino acid sequence of the point mutant NY-ESO peptide 6L (Fig. 127).

SEQ ID NO 127: amino acid sequence of the point mutant NY-ESO peptide 7F (Fig. 128).

SEQ ID NO 128: amino acid sequence of the point mutant NY-ESO peptide 8A (Fig. 129).

SEQ ID NO 129: amino acid sequence of the point mutant peptide 9A NY-ESO (Fig. 130).

SEQ ID NO 130: amino acid sequence of gp100 peptide (Fig. 131).

SEQ ID NO 131: amino acid sequence of the homologous peptide DOLPP1 (Fig. 132).

SEQ ID NO 132: amino acid sequence of the homologous peptide IL20RB (Fig. 133).

SEQ ID NO 133: amino acid sequence of the homologous peptide PRKD2 (Fig. 134).

SEQ ID NO 134: amino acid sequence of the homologous peptide of CD163 (Fig. 135).

SEQ ID NO 135: amino acid sequence of the homologous peptide P2RY8 (Fig. 136).

SEQ ID NO 136: amino acid sequence of C3E-7034 (Fig. 137).

SEQ ID NO 137: amino acid sequence of C3E-7036 (Fig. 138).

SEQ ID NO 138: amino acid sequence of C3E-7085 (Fig. 139).

SEQ ID NO 139: amino acid sequence of C3E-7088 (Fig. 140).

SEQ ID NO 140: amino acid sequence of C3E-7093 (Fig. 141).

SEQ ID NO 141: CDRH1 heavy chain C3E-7085 (Fig. 142).

SEQ ID NO 142: CDRH2 of heavy chain C3E-7085 (FIG. 142).

SEQ ID NO 143: CDRH3 of heavy chain C3E-7085 (Fig. 142).

SEQ ID NO 144: CDRL1 light chain C3E-7085 (Fig. 142).

SEQ ID NO 145: CDRL2 light chain C3E-7085 (Fig. 142).

SEQ ID NO 146: CDRL3 light chain C3E-7085 (Fig. 142).

SEQ ID NO 147: amino acid sequence of C3E-7078 (Fig. 143).

SEQ ID NO 148: nucleotide sequence of NYF-0014-HC2 (Fig. 144).

SEQ ID NO 149: amino acid sequence of NYF-0014-HC2 (Fig. 145); NYA-0001: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 150: amino acid sequence of NYF-0082-HC2 (Fig. 146); NYA-0082: amino acids 21-266; and C3E-7085: amino acids 272-511.

SEQ ID NO 151: amino acid sequence of human CD3 ε (Fig. 147).

SEQ ID NO 152: nucleotide sequence of full-length NYZ-0038-HC2 (Fig. 148).

SEQ ID NO 153: nucleotide sequence of full-length NYZ-0082-HC2 (Fig. 149).

SEQ ID NO 154: nucleotide sequence of full-length NYZ-0083-HC2 (Fig. 150).

SEQ ID NO 155: amino acid sequence of full-length NYZ-0038-HC2 (Fig. 151).

SEQ ID NO 156: amino acid sequence of full-length NYZ-0082-HC2 (Fig. 152).

SEQ ID NO 157: amino acid sequence of full-length NYZ-0083-HC2 (Fig. 153).

SEQ ID NO 158: nucleotide sequence of full-length NYZ-1010-HC2 (Fig. 154).

SEQ ID NO 159: nucleotide sequence of full-length C3E-7085-LC (Fig. 155).

SEQ ID NO 160: amino acid sequence of full-length NYZ-1010-HC2 (Fig. 156).

SEQ ID NO 161: amino acid sequence of full-length C3E-7085-LC (Fig. 157).

SEQ ID NO 162: amino acid sequence of the peptide linker (Fig. 161).

SEQ ID NO 163: nucleotide sequence of full-length NYA-3061 (Fig. 164).

SEQ ID NO 164: amino acid sequence of full-length NYA-3061 (Fig. 165).

SEQ ID NO 165: nucleotide sequence of full-length NYC-0005 (Fig. 166).

SEQ ID NO 166: amino acid sequence of full-length NYC-0005 (Fig. 167).

SEQ ID NO 167: nucleotide sequence of full-length NYC-0006 (Fig. 168).

SEQ ID NO 168: amino acid sequence of full-length NYC-0006 (Fig. 169).

SEQ ID NO 169: nucleotide sequence of full-length NYC-0007 (Fig. 170).

SEQ ID NO 170: amino acid sequence of full-length NYC-0007 (Fig. 171).

SEQ ID NO 171: nucleotide sequence of full-length NYC-0008 (Fig. 172).

SEQ ID NO 172: amino acid sequence of full-length NYC-0008 (Fig. 173).

SEQ ID NO 173: nucleotide sequence of full-length NYC-0009 (Fig. 174).

SEQ ID NO 174: amino acid sequence of full-length NYC-0009 (Fig. 175).

SEQ ID NO 175: nucleotide sequence of full-length NYC-0010 (Fig. 176).

SEQ ID NO 176: amino acid sequence of full-length NYC-0010 (Fig. 177).

SEQ ID NO 177: nucleotide sequence of full-length HC-h (Fig. 178).

SEQ ID NO 178: amino acid sequence of full-length HC-h (Fig. 179).

SEQ ID NO 179: nucleotide sequence of full-length NYD-2047-HC-k (Fig. 180).

SEQ ID NO 180: amino acid sequence of full-length NYD-2047-HC-k (Fig. 181).

SEQ ID NO 181: nucleotide sequence of full-length NYD-2061-HC-k (Fig. 182).

SEQ ID NO 182: amino acid sequence of full-length NYD-2061-HC-k (Fig. 183).

SEQ ID NO 183: nucleotide sequence of full-length NYD-3061-HC-k (Fig. 184).

SEQ ID NO 184: amino acid sequence of full-length NYD-3061-HC-k (Fig. 185).

SEQ ID NO 185: nucleotide sequence of full-length NYC-0011-HC-k (Fig. 186).

SEQ ID NO 186: amino acid sequence of full-length NYC-0011-HC-k (Fig. 187).

SEQ ID NO 187: nucleotide sequence of full-length NYC-0012-HC-k (Fig. 188).

SEQ ID NO 188: amino acid sequence of full-length NYC-0012-HC-k (Fig. 189).

SEQ ID NO 189: nucleotide sequence of full-length NYC-0013-HC-k (Fig. 190).

SEQ ID NO 190: amino acid sequence of full-length NYC-0013-HC-k (Fig. 191).

SEQ ID NO 191: nucleotide sequence of full-length NYC-0014-HC-k (Fig. 192).

SEQ ID NO 192: amino acid sequence of full-length NYC-0014-HC-k (Fig. 193).

SEQ ID NO 193: nucleotide sequence of full-length NYC-0015-HC-k (Fig. 194).

SEQ ID NO 194: amino acid sequence of full-length NYC-0015-HC-k (Fig. 195).

SEQ ID NO 195: nucleotide sequence of full-length NYC-0016-HC-k (Fig. 196).

SEQ ID NO 196: amino acid sequence of full-length NYC-0016-HC-k (Fig. 197).

SEQ ID NO 197: amino acid sequence of full-length NYZ-1007-HC (Fig. 198).

SEQ ID NO 198: amino acid sequence of full-length NYZ-1017-HC (Fig. 199).

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SEQUENCE LIST

<110> DAIICHI SANKYO COMPANY LIMITED

MIE UNIVERSITY

<120> BISPECIFIC ANTIBODIES

<130>PH-8759-PCT

<150> JP 2020-061476

<151> 2020-03-30

<160> 198

<170> PatentIn version 3.5

<210> 1

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> NY-ESO Peptide Sequence

<400> 1

Ser Leu Leu Met Trp Ile Thr Gln Cys

1 5

<210> 2

<211> 10

<212> PROTEIN

<213> Artificial

<220>

<223> Peptide sequence of MAGEC-1

<400> 2

Ile Leu Phe Gly Ile Ser Leu Arg Glu Val

1 5 10

<210> 3

<211> 26

<212> DNA

<213> Artificial

<220>

<223> Primer for scFv sequence analysis

<400> 3

ctcttcgcta ttacgccagc tggcga 26

<210> 4

<211> 30

<212> DNA

<213> Artificial

<220>

<223> Primer for scFv sequence analysis

<400> 4

ataacaattt cacacaggaa acagctatga 30

<210> 5

<211> 360

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the variable region of the heavy chain

NYA-0001

<400> 5

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 60

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 120

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 180

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 240

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 300

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 360

<210> 6

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the variable region of the heavy chain

NYA-0001

<400> 6

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn Met Val Tyr

65 70 75 80

Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 7

<211> 333

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the light chain variable region

NYA-0001

<400> 7

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 60

tcctgctctg gaagcagctc caacattggg aataattatg tatcctggta ccagcagctc 120

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 180

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 240

actggggacg aggccgatta ttactgcgga acatgggata gcagcctgag tgccccttgg 300

gtgttcggcg gagggaccaa ggtcaccgtc cta 333

<210> 8

<211> 111

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the light chain variable region

NYA-0001

<400> 8

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln

1 5 10 15

Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn

20 25 30

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln

65 70 75 80

Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

85 90 95

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 9

<211> 360

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the variable region of the heavy chain

NYA-0060

<400> 9

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 60

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 120

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 180

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 240

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 300

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 360

<210> 10

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the variable region of the heavy chain

NYA-0060

<400> 10

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn Met Val Tyr

65 70 75 80

Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 11

<211> 333

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the light chain variable region

NYA-0060

<400> 11

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 60

tcctgctctg gaagcagctc caacattggg aataattatg tatcctggta ccagcagctc 120

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 180

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 240

actggggacg aggccgatta ttactgcgga acatgggata gcagcctgag tgccccttgg 300

gtgttcggcg gagggaccaa ggtcaccgtc cta 333

<210> 12

<211> 111

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the light chain variable region

NYA-0060

<400> 12

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln

1 5 10 15

Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn

20 25 30

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln

65 70 75 80

Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

85 90 95

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 13

<211> 360

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the variable region of the heavy chain

NYA-0068

<400> 13

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 60

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 120

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 180

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 240

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 300

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 360

<210> 14

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the variable region of the heavy chain

NYA-0068

<400> 14

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn Met Val Tyr

65 70 75 80

Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 15

<211> 333

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the light chain variable region

NYA-0068

<400> 15

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 60

tcctgctctg gaagcagctc caacattggg aattggtatg tatcctggta ccagcagctc 120

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 180

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 240

actggggacg aggccgatta ttactgcgga acatgggata gcagcctgag tgccccttgg 300

gtgttcggcg gagggaccaa ggtcaccgtc cta 333

<210> 16

<211> 111

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the light chain variable region

NYA-0068

<400> 16

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln

1 5 10 15

Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

20 25 30

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln

65 70 75 80

Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

85 90 95

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 17

<211> 360

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the variable region of the heavy chain

NYA-0082

<400> 17

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 60

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 120

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 180

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 240

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 300

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 360

<210> 18

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the variable region of the heavy chain

NYA-0082

<400> 18

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn Met Val Tyr

65 70 75 80

Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 19

<211> 333

<212> DNA

<213> Artificial

<220>

<223> Nucleotide sequence of the light chain variable region

NYA-0082

<400> 19

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 60

tcctgctctg gaagcagctc caacattggg aataattatg tatcctggta ccagcggctc 120

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 180

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 240

actggggacg aggccgatta ttactgcgga tcatgggata gcagcctgag tgccccttgg 300

gtgttcggcg gagggaccaa ggtcaccgtc cta 333

<210> 20

<211> 111

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the light chain variable region

NYA-0082

<400> 20

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln

1 5 10 15

Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn

20 25 30

Tyr Val Ser Trp Tyr Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln

65 70 75 80

Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Ser Trp Asp Ser Ser Leu

85 90 95

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 21

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1163

<400> 21

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 22

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2023

<400> 22

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 23

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2027

<400> 23

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggatcatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 24

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1143

<400> 24

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 25

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2143

<400> 25

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc ggctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 26

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1163

<400> 26

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 27

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2023

<400> 27

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 28

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2027

<400> 28

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Ser Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 29

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1143

<400> 29

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 30

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2143

<400> 30

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 31

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1154

<400> 31

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tgcactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggccgttt atcactgtgc gagagggagt 360

agtaatcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc ggctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattactact gcggagcatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 32

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1154

<400> 32

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Asn His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Ala Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 33

<211> 290

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the truncated form

HLA-A*0201(UniProtKB-P01892)

<400> 33

Met Ser His Ser Met Arg Tyr Phe Phe Thr Ser Val Ser Arg Pro Gly

1 5 10 15

Arg Gly Glu Pro Arg Phe Ile Ala Val Gly Tyr Val Asp Asp Thr Gln

20 25 30

Phe Val Arg Phe Asp Ser Asp Ala Ala Ser Gln Arg Met Glu Pro Arg

35 40 45

Ala Pro Trp Ile Glu Gln Glu Gly Pro Glu Tyr Trp Asp Gly Glu Thr

50 55 60

Arg Lys Val Lys Ala His Ser Gln Thr His Arg Val Asp Leu Gly Thr

65 70 75 80

Leu Arg Gly Tyr Tyr Asn Gln Ser Glu Ala Gly Ser His Thr Val Gln

85 90 95

Arg Met Tyr Gly Cys Asp Val Gly Ser Asp Trp Arg Phe Leu Arg Gly

100 105 110

Tyr His Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Ile Ala Leu Lys Glu

115 120 125

Asp Leu Arg Ser Trp Thr Ala Ala Asp Met Ala Ala Gln Thr Thr Lys

130 135 140

His Lys Trp Glu Ala Ala His Val Ala Glu Gln Leu Arg Ala Tyr Leu

145 150 155 160

Glu Gly Thr Cys Val Glu Trp Leu Arg Arg Tyr Leu Glu Asn Gly Lys

165 170 175

Glu Thr Leu Gln Arg Thr Asp Ala Pro Lys Thr His Met Thr His His

180 185 190

Ala Val Ser Asp His Glu Ala Thr Leu Arg Cys Trp Ala Leu Ser Phe

195 200 205

Tyr Pro Ala Glu Ile Thr Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln

210 215 220

Thr Gln Asp Thr Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr

225 230 235 240

Phe Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Gln Glu Gln Arg

245 250 255

Tyr Thr Cys His Val Gln His Glu Gly Leu Pro Lys Pro Leu Thr Leu

260 265 270

Arg Trp Glu Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp

275 280 285

His Glu

290

<210> 34

<211> 100

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of beta 2-microglobulin

<400> 34

Met Ile Gln Arg Thr Pro Lys Ile Gln Val Tyr Ser Arg His Pro Ala

1 5 10 15

Glu Asn Gly Lys Ser Asn Phe Leu Asn Cys Tyr Val Ser Gly Phe His

20 25 30

Pro Ser Asp Ile Glu Val Asp Leu Leu Lys Asn Gly Glu Arg Ile Glu

35 40 45

Lys Val Glu His Ser Asp Leu Ser Phe Ser Lys Asp Trp Ser Phe Tyr

50 55 60

Leu Leu Tyr Tyr Thr Glu Phe Thr Pro Thr Glu Lys Asp Glu Tyr Ala

65 70 75 80

Cys Arg Val Asn His Val Thr Leu Ser Gln Pro Lys Ile Val Lys Trp

85 90 95

Asp Arg Asp Met

100

<210> 35

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2035

<400> 35

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gaaggtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 36

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2035

<400> 36

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Lys Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 37

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-1143-VH01

<400> 37

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 38

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-1143-VH02

<400> 38

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 39

<211> 120

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-1143-VH03

<400> 39

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 40

<211> 111

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-1143-VL01

<400> 40

Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

20 25 30

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

85 90 95

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 41

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2044

<400> 41

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgactcgcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 42

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2045

<400> 42

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgactcgcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 43

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2047

<400> 43

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

caagtacagc tggtggaatc cggtggaggc gtggtccagc cgggacgcag cttgagactg 120

tcctgcgctg catctggctt ttccatacga tcctacgata tgcactgggt tcgccaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattac 240

gcggattcag tgaaggggcg gttcacaatt tcacgggaca attcaaagaa taccttgtat 300

ctccagatgt cttcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 44

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2048

<400> 44

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

caagttcaac tcgtggaatc tggtggaggg gtggtacaac caggccggtc acttagactg 120

tcctgcgctg cgagtggatt ttccatcaga tcttacgaca tgcactgggt tcgccaagca 180

cccggaaagg gtttggaatg ggtggctacg atctcctacg atggatccca gaagtattac 240

gccgacagcg tcaagggccg atttacaata tcacgggaca attcaaagaa taccttgtat 300

ctccagatgt cttcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 45

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2060

<400> 45

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 46

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-2061

<400> 46

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 47

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2044

<400> 47

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 48

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2045

<400> 48

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 49

<211> 246

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-0082

<400> 49

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile Arg Ser Tyr

20 25 30

Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn Met Val Tyr

65 70 75 80

Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val Tyr His Cys

85 90 95

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

115 120 125

Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser

130 135 140

Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser Cys Ser Gly Ser

145 150 155 160

Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr Gln Arg Leu Pro

165 170 175

Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser

180 185 190

Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr

195 200 205

Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys

210 215 220

Gly Ser Trp Asp Ser Ser Leu Ser Ala Pro Trp Val Phe Gly Gly Gly

225 230 235 240

Thr Lys Val Thr Val Leu

245

<210> 50

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2047

<400> 50

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 51

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2048

<400> 51

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 52

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2060

<400> 52

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 53

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-2061

<400> 53

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 54

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-0001-CDRH1

<400> 54

Gly Phe Ser Ile Arg Ser Tyr Asp

1 5

<210> 55

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-0001-CDRH2

<400> 55

Ile Ser Tyr Asp Gly Ser Gln Lys

1 5

<210> 56

<211> 13

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence NYA-0001-CDRH3

<400> 56

Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile

1 5 10

<210> 57

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-0001-CDRL1

<400> 57

Ser Ser Asn Ile Gly Asn Asn Tyr

1 5

<210> 58

<211> 3

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-0001-CDRL2

<400> 58

Asp Asn Asn

1

<210> 59

<211> 12

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-0001-CDRL3

<400> 59

Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

1 5 10

<210> 60

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-2023-CDRL1

<400> 60

Ser Ser Asn Ile Gly Asn Trp Tyr

1 5

<210> 61

<211> 12

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-2027-CDRL3

<400> 61

Gly Ser Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

1 5 10

<210> 62

<211> 13

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-1154-CDRH3

<400> 62

Ala Arg Gly Ser Ser Asn His Tyr Glu Ala Phe Asp Ile

1 5 10

<210> 63

<211> 12

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-1154-CDRL3

<400> 63

Gly Ala Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

1 5 10

<210> 64

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of NYA-0035-CDRL1

<400> 64

Ser Ser Asn Ile Gly Asn Trp Lys

1 5

<210> 65

<211> 867

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0003

<400> 65

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggc 60

gaagtccagc tgttggagtc aggtggcgga ttggtgcaac ctggcgggtc actgaggctg 120

agttgtgcag ctagcggctt cacattctct acgtatcaga tgagctgggt gagacaggct 180

caggaaagg gtctggaatg ggtcagcggg attgtgtcta gcggtggctc cactgcctat 240

gccgatagcg taaaaggccg ctttacgatc tctcgggaca actctaagaa cacactctat 300

ctgcagatga attcccttag agccgaagat accgccgtgt actactgtgc tggggaactg 360

ctcccgtatt acggtatgga cgtttggggc caaggcacca ctgtcacagt gagttccggt 420

ggaggcgggt caggcggagg cggtagtgga ggtggaggat cacaaagcga gctgacacag 480

cctaggtccg tatccggaag tccagggcag agcgtcacca tcagctgcac tggcacctct 540

cgagatgtcg gcggatacaa ctacgtgtct tggtatcagc agcatcccgg caaagcgccc 600

aaactcatca tacacgacgt gattgagcgg tccagtgggg ttcccgatcg tttcagcggg 660

tcaaagtccg gaaataccgc aagcctgacc atttctgggc ttcaggcaga ggatgaggct 720

gactactact gctggtcctt tgccgggagc tattacgtgt ttgggacagg gactgacgtg 780

actgttctgg gcgcggccgc aggtgcaggt ggtgattaca aagatgatga cgataaaggt 840

gcagcggcgc atcaccatca tcaccac 867

<210> 66

<211> 867

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0004

<400> 66

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggc 60

gaagtccagc tgttggagtc aggtggcgga ttggtgcaac ctggcgggtc actgaggctg 120

agttgtgcag ctagcggctt cacattctct acgtatcaga tgagctgggt gagacaggct 180

caggaaagg gtctggaatg ggtcagcggg attgtgtcta gcggtggctc cactgcctat 240

gccgatagcg taaaaggccg ctttacgatc tctcgggaca actctaagaa cacactctat 300

ctgcagatga attcccttag agccgaagat accgccgtgt actactgtgc tggggaactg 360

ctcccgtatt acggtatgga cgtttggggc caaggcacca ctgtcacagt gagttccggt 420

ggaggcgggt caggcggagg cggtagtgga ggtggaggat cacaaagcga gctgacacag 480

cctaggtccg tatccggaag tccagggcag agcgtcacca tcagctgcac tggcaccgag 540

cgagatgtcg gcggatacaa ctacgtgtct tggtatcagc agcatcccgg caaagcgccc 600

aaactcatca tacacgacgt gattgagcgg tccagtgggg ttcccgatcg tttcagcggg 660

tcaaagtccg gaaataccgc aagcctgacc atttctgggc ttcaggcaga ggatgaggct 720

gactactact gctggtcctt tgccgggggc tattacgtgt ttgggacagg gactgacgtg 780

actgttctgg gcgcggccgc aggtgcaggt ggtgattaca aagatgatga cgataaaggt 840

gcagcggcgc atcaccatca tcaccac 867

<210> 67

<211> 289

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0003

<400> 67

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Ser Thr Tyr Gln Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Gly Ile Val Ser Ser Gly Gly Ser Thr Ala Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr Tyr Cys Ala Gly Glu Leu Leu Pro Tyr Tyr Gly Met Asp Val

115 120 125

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser

130 135 140

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Glu Leu Thr Gln

145 150 155 160

Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val Thr Ile Ser Cys

165 170 175

Thr Gly Thr Ser Arg Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Ile Ile His Asp Val Ile

195 200 205

Glu Arg Ser Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Trp Ser Phe Ala Gly Ser Tyr Tyr Val Phe Gly Thr

245 250 255

Gly Thr Asp Val Thr Val Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp

260 265 270

Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His His His

275 280 285

His

<210> 68

<211> 289

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0004

<400> 68

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Ser Thr Tyr Gln Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Gly Ile Val Ser Ser Gly Gly Ser Thr Ala Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr Tyr Cys Ala Gly Glu Leu Leu Pro Tyr Tyr Gly Met Asp Val

115 120 125

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser

130 135 140

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Glu Leu Thr Gln

145 150 155 160

Pro Arg Ser Val Ser Gly Ser Pro Gly Gln Ser Val Thr Ile Ser Cys

165 170 175

Thr Gly Thr Glu Arg Asp Val Gly Gly Tyr Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln His Pro Gly Lys Ala Pro Lys Leu Ile Ile His Asp Val Ile

195 200 205

Glu Arg Ser Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Trp Ser Phe Ala Gly Gly Tyr Tyr Val Phe Gly Thr

245 250 255

Gly Thr Asp Val Thr Val Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp

260 265 270

Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His His His

275 280 285

His

<210> 69

<211> 876

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-0001

<400> 69

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg cgcggccgca ggtgcaggtg gtgattacaa agatgatgac 840

gataaaggtg cagcggcgca tcaccatcat caccac 876

<210> 70

<211> 292

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-0001

<400> 70

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Ala Ala Ala Gly Ala

260 265 270

Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala Ala His His

275 280 285

His His His His

290

<210> 71

<211> 738

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of HC1

<400> 71

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgac 60

aaaactcaca catgcccacc ctgcccagca cctgaagccg cagggggacc ctcagtcttc 120

ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 180

gtggtggtgt cagtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 240

gtggaggtgc ataatgccaa gacaaagccc cgggaggagc agtacaacag cacgtaccgg 300

gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 360

aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggc 420

cagccccggg aaccacaggt gtacgtgctg cccccatccc gggacgagct gaccaagaac 480

caggtcagcc tgctgtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 540

gagagcaatg gccagcccga gaacaactac ctgacctggc ctcccgtgct ggactccgac 600

ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcagggcaac 660

gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacaccca gaagagcctc 720

tccctgtctc ccggcaag 738

<210> 72

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0016-HC2

<400> 72

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 73

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0019-HC2

<400> 73

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc ggctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 74

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0022-HC2

<400> 74

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 75

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0023-HC2

<400> 75

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 76

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0027-HC2

<400> 76

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggatcatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 77

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0035-HC2

<400> 77

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gaaggtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 78

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0044-HC2

<400> 78

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgactcgcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 79

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0045-HC2

<400> 79

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgactcgcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 80

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0047-HC2

<400> 80

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

caagtacagc tggtggaatc cggtggaggc gtggtccagc cgggacgcag cttgagactg 120

tcctgcgctg catctggctt ttccatacga tcctacgata tgcactgggt tcgccaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattac 240

gcggattcag tgaaggggcg gttcacaatt tcacgggaca attcaaagaa taccttgtat 300

ctccagatgt cttcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 81

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0048-HC2

<400> 81

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

caagttcaac tcgtggaatc tggtggaggg gtggtacaac caggccggtc acttagactg 120

tcctgcgctg cgagtggatt ttccatcaga tcttacgaca tgcactgggt tcgccaagca 180

cccggaaagg gtttggaatg ggtggctacg atctcctacg atggatccca gaagtattac 240

gccgacagcg tcaagggccg atttacaata tcacgggaca attcaaagaa taccttgtat 300

ctccagatgt cttcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 82

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0060-HC2

<400> 82

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 83

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0061-HC2

<400> 83

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 84

<211> 246

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of HC1

<400> 84

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

20 25 30

Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

35 40 45

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ser

50 55 60

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

65 70 75 80

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

85 90 95

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

100 105 110

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

115 120 125

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

130 135 140

Pro Gln Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn

145 150 155 160

Gln Val Ser Leu Leu Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

165 170 175

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Leu Thr

180 185 190

Trp Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

195 200 205

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

210 215 220

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

225 230 235 240

Ser Leu Ser Pro Gly Lys

245

<210> 85

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0016-HC

<400> 85

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 86

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0019-HC2

<400> 86

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 87

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0022-HC2

<400> 87

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 88

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0023-HC2

<400> 88

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 89

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0027-HC2

<400> 89

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Ser Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 90

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0035-HC2

<400> 90

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Lys Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 91

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0044-HC2

<400> 91

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 92

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0045-HC2

<400> 92

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 93

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0047-HC2

<400> 93

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 94

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0048-HC2

<400> 94

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 95

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0060-HC2

<400> 95

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 96

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0061-HC2

<400> 96

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 97

<211> 1404

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-0001-Fab-HC1-k

deletion

<400> 97

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgag 60

gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gacaccctcc 120

tgttcagcgt ctggattctc catccgtagt tatgatatac actgggtccg ccaggctcca 180

ggcaaggggc tagagtgggt ggccactata tcatatgatg gaagtcagaa gtacttcgca 240

gactccgtga agggccgatt taccatcttc agagacgaat cggagaacat ggtgtatctg 300

caaatgagcg gcctgagagt tgaggacacg gctgtttatc actgtgcgag agggagtagt 360

ggtcattatg aggcttttga tatctggggc caagggacaa tggtcaccgt ctcttcagcc 420

tccaccaagg gcccaagcgt cttccccctg gcaccctcct ccaagagcac ctctggcggc 480

acagccgccc tgggctgcct ggtcaaggac tacttccccg aacccgtgac cgtgagctgg 540

aactcaggcg ccctgaccag cggcgtgcac accttccccg ctgtcctgca gtcctcagga 600

ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 660

atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagagagt tgagcccaaa 720

tcttgtgaca aaactcacac atgcccaccc tgcccagcac ctgaagccgc aggggaccc 780

tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 840

gtcacatgcg tggtggtgtc agtgagccac gaagaccctg aggtcaagtt caactggtac 900

gtggacggcg tggaggtgca taatgccaag acaaagcccc gggaggagca gtacaacagc 960

acgtaccggg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 1020

tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 1080

gccaaaggcc agccccggga accacaggtg tacgtgctgc ccccatcccg ggacgagctg 1140

accaagaacc aggtcagcct gctgtgcctg gtcaaaggct tctatcccag cgacatcgcc 1200

gtggagtggg agagcaatgg ccagcccgag aacaactacc tgacctggcc tcccgtgctg 1260

gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 1320

cagggcaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacccag 1380

aagagcctct ccctgtctcc cggc 1404

<210> 98

<211> 711

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-0001-LC

<400> 98

atggtgctgc agacccaggt gttcatctcc ctgctgctgt ggatctccgg cgcgtacggc 60

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 120

tcctgctctg gaagcagctc caacattggg aataattatg tatcctggta ccagcagctc 180

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 240

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 300

actggggacg aggccgatta ttactgcgga acatgggata gcagcctgag tgccccttgg 360

gtgttcggcg gagggaccaa ggtcaccgtc ctaggccagc ctaaggctgc ccctagcgtg 420

accctgttcc ctccttccag cgaggagctt caagctaaca aggccaccct ggtgtgtctt 480

atctctgact tctaccctgg cgctgtgacc gtggcctgga aggctgacag ctcccctgtg 540

aaggccggag tggagaccac cacacctagc aagcagtcta acaacaagta cgctgccagc 600

tcctacctga gccttacccc tgagcagtgg aagtctcaca gaagctactc ctgtcaagtg 660

acccacgagg gcagcaccgt ggagaagacc gtggctccta ccgagtgttc c 711

<210> 99

<211> 468

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-0001-Fab-HC1-k

deletion

<400> 99

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln

20 25 30

Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile

35 40 45

Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

50 55 60

Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala

65 70 75 80

Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn

85 90 95

Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val

100 105 110

Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile

115 120 125

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly

130 135 140

Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

145 150 155 160

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

165 170 175

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

180 185 190

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

195 200 205

Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

210 215 220

Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys

225 230 235 240

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala

245 250 255

Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

260 265 270

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ser Val

275 280 285

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

290 295 300

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

305 310 315 320

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

325 330 335

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

340 345 350

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

355 360 365

Gln Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln

370 375 380

Val Ser Leu Leu Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

385 390 395 400

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Leu Thr Trp

405 410 415

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

420 425 430

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

435 440 445

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

450 455 460

Leu Ser Pro Gly

465

<210> 100

<211> 237

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-0001-LC

<400> 100

Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser

1 5 10 15

Gly Ala Tyr Gly Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala

20 25 30

Ala Pro Gly Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn

35 40 45

Ile Gly Asn Asn Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala

50 55 60

Pro Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro

65 70 75 80

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile

85 90 95

Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp

100 105 110

Asp Ser Ser Leu Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val

115 120 125

Thr Val Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro

130 135 140

Pro Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu

145 150 155 160

Ile Ser Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp

165 170 175

Ser Ser Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln

180 185 190

Ser Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu

195 200 205

Gln Trp Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly

210 215 220

Ser Thr Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

225 230 235

<210> 101

<211> 1404

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1143-Fab-HC1-k

deletion

<400> 101

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgag 60

gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gacaccctcc 120

tgttcagcgt ctggattctc catccgtagc tatgatatac actgggtccg cctggctcca 180

ggcaaggggc tagagtgggt ggccactata tcatatgatg gaagtcagaa gtacttcgca 240

gactccgtga agggccgatt taccatcttc agagacgaat cggagaacat ggtgtatctg 300

caaatgagcg gcctgagagt tgaggacacg gctgtttatc actgtgcgag agggagtagt 360

ggtcattatg aggcttttga tatctggggc caagggacaa tggtcaccgt ctcttcagcc 420

tccaccaagg gcccaagcgt cttccccctg gcaccctcct ccaagagcac ctctggcggc 480

acagccgccc tgggctgcct ggtcaaggac tacttccccg aacccgtgac cgtgagctgg 540

aactcaggcg ccctgaccag cggcgtgcac accttccccg ctgtcctgca gtcctcagga 600

ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 660

atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagagagt tgagcccaaa 720

tcttgtgaca aaactcacac atgcccaccc tgcccagcac ctgaagccgc aggggaccc 780

tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 840

gtcacatgcg tggtggtgtc agtgagccac gaagaccctg aggtcaagtt caactggtac 900

gtggacggcg tggaggtgca taatgccaag acaaagcccc gggaggagca gtacaacagc 960

acgtaccggg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 1020

tacaagtgca aggtctccaa caaagccctc ccagccccca tcgagaaaac catctccaaa 1080

gccaaaggcc agccccggga accacaggtg tacgtgctgc ccccatcccg ggacgagctg 1140

accaagaacc aggtcagcct gctgtgcctg gtcaaaggct tctatcccag cgacatcgcc 1200

gtggagtggg agagcaatgg ccagcccgag aacaactacc tgacctggcc tcccgtgctg 1260

gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 1320

cagggcaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacccag 1380

aagagcctct ccctgtctcc cggc 1404

<210> 102

<211> 711

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1143-LC

<400> 102

atggtgctgc agacccaggt gttcatctcc ctgctgctgt ggatctccgg cgcgtacggc 60

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 120

tcctgctctg gaagcagctc caacattggg aattggtatg tatcctggta ccagcagctc 180

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 240

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 300

actggggacg aggccgatta ttactgcgga acatgggata gcagcctgag tgccccttgg 360

gtgttcggcg gagggaccaa ggtcaccgtc ctaggccagc ctaaggctgc ccctagcgtg 420

accctgttcc ctccttccag cgaggagctt caagctaaca aggccaccct ggtgtgtctt 480

atctctgact tctaccctgg cgctgtgacc gtggcctgga aggctgacag ctcccctgtg 540

aaggccggag tggagaccac cacacctagc aagcagtcta acaacaagta cgctgccagc 600

tcctacctga gccttacccc tgagcagtgg aagtctcaca gaagctactc ctgtcaagtg 660

acccacgagg gcagcaccgt ggagaagacc gtggctccta ccgagtgttc c 711

<210> 103

<211> 1476

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of C3E-7085-HC2-k deletion

<400> 103

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaagtgcagc tggtggaatc cggggggggc ctggtgcagc ctggggggag cctgagactg 120

agttgtgccg cctctggggt gacatttaac tactatggca tgtcttggat ccgccaggca 180

cctggaaagg gcctggagtg ggtggccagc atcactaggt ccggcgggcg aatctactat 240

cccgacagcg tcaagggcag gttcacaatt tcccgcgaga acacacagaa aactctgtac 300

ctgcagatga atagcctgag agccgaagat acagctgtgt actattgcac tctggacggc 360

agggatgggt gggtcgccta ttggggggcag ggaaccctgg tgacagtcag ctccggagga 420

ggaggatctg gcggaggagg cagtggggga ggcgggtcaa actttatgct cactcagccg 480

tcctctgttt ctggcgtacc tggccaacgg gtgaccatta gctgtacggg taataccggg 540

aatatcgggt ctaactacgt gaactggtat cagcagcttc cagggacagc tcccaagttg 600

ctgatctatc gcgacgacaa aagaccctca ggggtccctg accgatttag tggcagcaaa 660

agcggtactt ccgcttccct ggcgataacc ggctttcagg ccgaagatga ggcagactac 720

tattgccagt catattccag cggcttcatc ttcggaggcg gaactaagct gacagtgttg 780

gcagccgagc ccaaatcttc tgacaaaact cacacatgcc caccctgccc agcacctgaa 840

gccgcagggg gaccctcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc 900

tcccggaccc ctgaggtcac atgcgtggtg gtgtcagtga gccacgaaga ccctgaggtc 960

aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccccggggag 1020

gagcagtaca acagcacgta ccgggtggtc agcgtcctca ccgtcctgca ccaggactgg 1080

ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag 1140

aaaaccatct ccaaagccaa aggccagccc cgggaaccac aggtgtacgt gtacccccca 1200

tcccgggacg agctgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat 1260

cccagcgaca tcgccgtgga gtgggagagc aatggccagc ccgagaacaa ctacaagacc 1320

acccctcccg tgctggactc cgacggctcc ttcgccctcg tgagcaagct caccgtggac 1380

aagagcaggt ggcagcaggg caacgtcttc tcatgctccg tgatgcatga ggctctgcac 1440

aaccactaca cccagaagag cctctccctg tctccc 1476

<210> 104

<211> 468

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1143-Fab-HC1-k

deletion

<400> 104

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln

20 25 30

Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser Ile

35 40 45

Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly Leu

50 55 60

Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe Ala

65 70 75 80

Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu Asn

85 90 95

Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala Val

100 105 110

Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp Ile

115 120 125

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly

130 135 140

Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

145 150 155 160

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

165 170 175

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

180 185 190

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

195 200 205

Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

210 215 220

Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys

225 230 235 240

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala

245 250 255

Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

260 265 270

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ser Val

275 280 285

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

290 295 300

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

305 310 315 320

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

325 330 335

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

340 345 350

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

355 360 365

Gln Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln

370 375 380

Val Ser Leu Leu Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

385 390 395 400

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Leu Thr Trp

405 410 415

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

420 425 430

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

435 440 445

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

450 455 460

Leu Ser Pro Gly

465

<210> 105

<211> 237

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1143-LC

<400> 105

Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser

1 5 10 15

Gly Ala Tyr Gly Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala

20 25 30

Ala Pro Gly Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn

35 40 45

Ile Gly Asn Trp Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala

50 55 60

Pro Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro

65 70 75 80

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile

85 90 95

Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp

100 105 110

Asp Ser Ser Leu Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val

115 120 125

Thr Val Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro

130 135 140

Pro Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu

145 150 155 160

Ile Ser Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp

165 170 175

Ser Ser Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln

180 185 190

Ser Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu

195 200 205

Gln Trp Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly

210 215 220

Ser Thr Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

225 230 235

<210> 106

<211> 493

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of C3E-7085-HC2-k deletion

<400> 106

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr

35 40 45

Phe Asn Tyr Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr

65 70 75 80

Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln

85 90 95

Lys Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr Tyr Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp

115 120 125

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

130 135 140

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro

145 150 155 160

Ser Ser Val Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr

165 170 175

Gly Asn Thr Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln

180 185 190

Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg

195 200 205

Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser

210 215 220

Ala Ser Leu Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr

225 230 235 240

Tyr Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys

245 250 255

Leu Thr Val Leu Ala Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr

260 265 270

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

275 280 285

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

290 295 300

Glu Val Thr Cys Val Val Val Ser Val Ser His Glu Asp Pro Glu Val

305 310 315 320

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

325 330 335

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

340 345 350

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

355 360 365

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

370 375 380

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro

385 390 395 400

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

405 410 415

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

420 425 430

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

435 440 445

Gly Ser Phe Ala Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

450 455 460

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

465 470 475 480

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

485 490

<210> 107

<211> 1497

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1143-HC1-k deletion

<400> 107

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagc agccgagccc aaatcttctg acaaaactca cacatgccca 840

ccctgcccag cacctgaagc cgcaggggga ccctcagtct tcctcttccc cccaaaaccc 900

aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt gtcagtgagc 960

cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt gcataatgcc 1020

aagacaaagc cccgggagga gcagtacaac agcacgtacc gggtggtcag cgtcctcacc 1080

gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc caacaaagcc 1140

ctcccagccc ccatcgagaa aaccatctcc aaagccaaag gccagccccg ggaaccacag 1200

gtgtacgtgc tgcccccatc ccgggacgag ctgaccaaga accaggtcag cctgctgtgc 1260

ctggtcaaag gcttctatcc cagcgacatc gccgtggagt ggggagagcaa tggccagccc 1320

gagaacaact acctgacctg gcctcccgtg ctggactccg acggctcctt cttcctctac 1380

agcaagctca ccgtggacaa gagcaggtgg cagcagggca acgtcttctc atgctccgtg 1440

atgcatgagg ctctgcacaa ccactacacc cagaagagcc tctccctgtc tcccggc 1497

<210> 108

<211> 499

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1143-HC1-k deletion

<400> 108

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Ala Ala Glu Pro Lys Ser

260 265 270

Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

275 280 285

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

290 295 300

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ser Val Ser

305 310 315 320

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

325 330 335

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

340 345 350

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

355 360 365

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

370 375 380

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

385 390 395 400

Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

405 410 415

Ser Leu Leu Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

420 425 430

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Leu Thr Trp Pro

435 440 445

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

450 455 460

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

465 470 475 480

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

485 490 495

Ser Pro Gly

<210> 109

<211> 2142

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of C3E-7085-NYA-1154-Fab-HC2-k

deletion

<400> 109

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaagtgcagc tggtggaatc cggggggggc ctggtgcagc ctggggggag cctgagactg 120

agttgtgccg cctctggggt gacatttaac tactatggca tgtcttggat ccgccaggca 180

cctggaaagg gcctggagtg ggtggccagc atcactaggt ccggcgggcg aatctactat 240

cccgacagcg tcaagggcag gttcacaatt tcccgcgaga acacacagaa aactctgtac 300

ctgcagatga atagcctgag agccgaagat acagctgtgt actattgcac tctggacggc 360

agggatgggt gggtcgccta ttggggggcag ggaaccctgg tgacagtcag ctccggagga 420

ggaggatctg gcggaggagg cagtggggga ggcgggtcaa actttatgct cactcagccg 480

tcctctgttt ctggcgtacc tggccaacgg gtgaccatta gctgtacggg taataccggg 540

aatatcgggt ctaactacgt gaactggtat cagcagcttc cagggacagc tcccaagttg 600

ctgatctatc gcgacgacaa aagaccctca ggggtccctg accgatttag tggcagcaaa 660

agcggtactt ccgcttccct ggcgataacc ggctttcagg ccgaagatga ggcagactac 720

tattgccagt catattccag cggcttcatc ttcggaggcg gaactaagct gacagtgttg 780

gggggaggcg gttcagaggt gcagctggtg gagtctgggg gaggcgtggt ccagcctggg 840

aggtccctga caccctcctg ttcagcgtct ggattctcca tccgtagtta tgatatgcac 900

tgggtccgcc aggctccagg caaggggcta gagtgggtgg ccactatatc atatgatgga 960

agtcagaagt acttcgcaga ctccgtgaag ggccgattta ccatcttcag agacgaatcg 1020

gagaacatgg tgtatctgca aatgagcggc ctgagagttg aggacacggc cgtttatcac 1080

tgtgcgagag ggagtagtaa tcattatgag gcttttgata tctggggcca agggacaatg 1140

gtcaccgtct cttcagcctc caccaagggc ccaagcgtct tccccctggc accctcctcc 1200

aagagcacct ctggcggcac agccgccctg ggctgcctgg tcaaggacta cttccccgaa 1260

cccgtgaccg tgagctggaa ctcaggcgcc ctgaccagcg gcgtgcacac cttccccgct 1320

gtcctgcagt cctcaggact ctactccctc agcagcgtgg tgaccgtgcc ctccagcagc 1380

ttgggcaccc agacctacat ctgcaacgtg aatcacaagc ccagcaacac caaggtggac 1440

aagagagttg agcccaaatc ttgtgacaaa actcacacat gcccaccctg cccagcacct 1500

gaagccgcag ggggaccctc agtcttcctc ttccccccaa aacccaagga caccctcatg 1560

atctcccgga cccctgaggt cacatgcgtg gtggtgtcag tgagccacga agaccctgag 1620

gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccccgg 1680

gagggagcagt acaacagcac gtaccgggtg gtcagcgtcc tcaccgtcct gcaccaggac 1740

tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc 1800

gagaaaacca tctccaaagc caaaggccag ccccgggaac cacaggtgta cgtgtacccc 1860

ccatcccggg acgagctgac caagaaccag gtcagcctga cctgcctggt caaaggcttc 1920

tatcccagcg acatcgccgt ggagtgggag agcaatggcc agcccgagaa caactacaag 1980

accacccctc ccgtgctgga ctccgacggc tccttcgccc tcgtgagcaa gctcaccgtg 2040

gacaagagca ggtggcagca gggcaacgtc ttctcatgct ccgtgatgca tgaggctctg 2100

cacaaccact acacccagaa gagcctctcc ctgtctcccg gc 2142

<210> 110

<211> 711

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-1154-LC

<400> 110

atggtgctgc agacccaggt gttcatctcc ctgctgctgt ggatctccgg cgcgtacggc 60

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 120

tcctgctctg gaagcagctc caacattggg aataattatg tatcctggta ccagcggctc 180

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 240

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 300

actggggacg aggccgatta ctactgcgga gcatgggata gcagcctgag tgccccttgg 360

gtgttcggcg gagggaccaa ggtcaccgtc ctaggccagc ctaaggctgc ccctagcgtg 420

accctgttcc ctccttccag cgaggagctt caagctaaca aggccaccct ggtgtgtctt 480

atctctgact tctaccctgg cgctgtgacc gtggcctgga aggctgacag ctcccctgtg 540

aaggccggag tggagaccac cacacctagc aagcagtcta acaacaagta cgctgccagc 600

tcctacctga gccttacccc tgagcagtgg aagtctcaca gaagctactc ctgtcaagtg 660

acccacgagg gcagcaccgt ggagaagacc gtggctccta ccgagtgttc c 711

<210> 111

<211> 735

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of OAA-HC1-k deletion

<400> 111

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgac 60

aaaactcaca catgcccacc ctgcccagca cctgaagccg cagggggacc ctcagtcttc 120

ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 180

gtggtggtgt cagtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 240

gtggaggtgc ataatgccaa gacaaagccc cgggaggagc agtacaacag cacgtaccgg 300

gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 360

aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggc 420

cagccccggg aaccacaggt gtacgtgctg cccccatccc gggacgagct gaccaagaac 480

caggtcagcc tgctgtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 540

gagagcaatg gccagcccga gaacaactac ctgacctggc ctcccgtgct ggactccgac 600

ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcagggcaac 660

gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacaccca gaagagcctc 720

tccctgtctc ccggc 735

<210> 112

<211> 714

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of C3E-7085-NYA-1154-Fab-

HC2-k deletion

<400> 112

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr

35 40 45

Phe Asn Tyr Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr

65 70 75 80

Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln

85 90 95

Lys Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr Tyr Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp

115 120 125

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

130 135 140

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro

145 150 155 160

Ser Ser Val Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr

165 170 175

Gly Asn Thr Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln

180 185 190

Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg

195 200 205

Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser

210 215 220

Ala Ser Leu Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr

225 230 235 240

Tyr Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys

245 250 255

Leu Thr Val Leu Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser

260 265 270

Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser

275 280 285

Ala Ser Gly Phe Ser Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln

290 295 300

Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly

305 310 315 320

Ser Gln Lys Tyr Phe Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe

325 330 335

Arg Asp Glu Ser Glu Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg

340 345 350

Val Glu Asp Thr Ala Val Tyr His Cys Ala Arg Gly Ser Ser Asn His

355 360 365

Tyr Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser

370 375 380

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser

385 390 395 400

Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

405 410 415

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

420 425 430

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr

435 440 445

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln

450 455 460

Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp

465 470 475 480

Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro

485 490 495

Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro

500 505 510

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

515 520 525

Cys Val Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn

530 535 540

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

545 550 555 560

Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

565 570 575

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

580 585 590

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

595 600 605

Gly Gln Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp

610 615 620

Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

625 630 635 640

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

645 650 655

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

660 665 670

Ala Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly

675 680 685

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

690 695 700

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

705 710

<210> 113

<211> 237

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-1154-LC

<400> 113

Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser

1 5 10 15

Gly Ala Tyr Gly Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala

20 25 30

Ala Pro Gly Gln Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn

35 40 45

Ile Gly Asn Asn Tyr Val Ser Trp Tyr Gln Arg Leu Pro Gly Thr Ala

50 55 60

Pro Lys Leu Leu Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro

65 70 75 80

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile

85 90 95

Thr Gly Leu Gln Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Ala Trp

100 105 110

Asp Ser Ser Leu Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val

115 120 125

Thr Val Leu Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro

130 135 140

Pro Ser Ser Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu

145 150 155 160

Ile Ser Asp Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp

165 170 175

Ser Ser Pro Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln

180 185 190

Ser Asn Asn Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu

195 200 205

Gln Trp Lys Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly

210 215 220

Ser Thr Val Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

225 230 235

<210> 114

<211> 245

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of OAA-HC1-k deletion

<400> 114

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

20 25 30

Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

35 40 45

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ser

50 55 60

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

65 70 75 80

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

85 90 95

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

100 105 110

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

115 120 125

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

130 135 140

Pro Gln Val Tyr Val Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn

145 150 155 160

Gln Val Ser Leu Leu Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

165 170 175

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Leu Thr

180 185 190

Trp Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

195 200 205

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

210 215 220

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

225 230 235 240

Ser Leu Ser Pro Gly

245

<210> 115

<211> 2232

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0010-HC2-k deletion

<400> 115

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tgcactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggccgttt atcactgtgc gagagggagt 360

agtaatcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc ggctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattactact gcggagcatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gc 2232

<210> 116

<211> 2232

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0004-HC2-k deletion

<400> 116

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaagtgcagc tggtggaatc cggggggggc ctggtgcagc ctggggggag cctgagactg 120

agttgtgccg cctctggggt gacatttaac tactatggca tgtcttggat ccgccaggca 180

cctggaaagg gcctggagtg ggtggccagc atcactaggt ccggcgggcg aatctactat 240

cccgacagcg tcaagggcag gttcacaatt tcccgcgaga acacacagaa aactctgtac 300

ctgcagatga atagcctgag agccgaagat acagctgtgt actattgcac tctggacggc 360

agggatgggt gggtcgccta ttggggggcag ggaaccctgg tgacagtcag ctccggagga 420

ggaggatctg gcggaggagg cagtggggga ggcgggtcaa actttatgct cactcagccg 480

tcctctgttt ctggcgtacc tggccaacgg gtgaccatta gctgtacggg taataccggg 540

aatatcgggt ctaactacgt gaactggtat cagcagcttc cagggacagc tcccaagttg 600

ctgatctatc gcgacgacaa aagaccctca ggggtccctg accgatttag tggcagcaaa 660

agcggtactt ccgcttccct ggcgataacc ggctttcagg ccgaagatga ggcagactac 720

tattgccagt catattccag cggcttcatc ttcggaggcg gaactaagct gacagtgttg 780

gggggaggcg gttcagaggt gcagctggtg gagtctgggg gaggcgtggt ccagcctggg 840

aggtccctga caccctcctg ttcagcgtct ggattctcca tccgtagtta tgatatgcac 900

tgggtccgcc aggctccagg caaggggcta gagtgggtgg ccactatatc atatgatgga 960

agtcagaagt acttcgcaga ctccgtgaag ggccgattta ccatcttcag agacgaatcg 1020

gagaacatgg tgtatctgca aatgagcggc ctgagagttg aggacacggc cgtttatcac 1080

tgtgcgagag ggagtagtaa tcattatgag gcttttgata tctggggcca agggacaatg 1140

gtcaccgtct cttcaggtgg aggcggttca ggcggaggtg gcagcggcgg tggcgggagt 1200

cagtctgtgt tgacgcagcc gccctcagtg tctgcggccc caggacagaa ggtcaccatc 1260

tcctgctctg gaagcagctc caacattggg aataattatg tatcctggta ccagcggctc 1320

cccaggaacag cccccaaact cctcatttat gacaataata agcgaccctc agggattcct 1380

gaccgattct ctggctccaa gtctggcacg tcagccaccc tgggcatcac cggactccag 1440

actggggacg aggccgatta ctactgcgga gcatgggata gcagcctgag tgccccttgg 1500

gtgttcggcg gagggaccaa ggtcaccgtc ctagcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gc 2232

<210> 117

<211> 2232

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0011-HC2-k deletion

<400> 117

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agctatgata tacactgggt ccgcctggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaattg gtatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gc 2232

<210> 118

<211> 744

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0010-HC2-k deletion

<400> 118

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Asn His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Ala Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly

740

<210> 119

<211> 744

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0004-HC2-k deletion

<400> 119

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr

35 40 45

Phe Asn Tyr Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr

65 70 75 80

Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln

85 90 95

Lys Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr Tyr Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp

115 120 125

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

130 135 140

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro

145 150 155 160

Ser Ser Val Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr

165 170 175

Gly Asn Thr Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln

180 185 190

Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg

195 200 205

Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser

210 215 220

Ala Ser Leu Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr

225 230 235 240

Tyr Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys

245 250 255

Leu Thr Val Leu Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser

260 265 270

Gly Gly Gly Val Val Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser

275 280 285

Ala Ser Gly Phe Ser Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln

290 295 300

Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly

305 310 315 320

Ser Gln Lys Tyr Phe Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe

325 330 335

Arg Asp Glu Ser Glu Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg

340 345 350

Val Glu Asp Thr Ala Val Tyr His Cys Ala Arg Gly Ser Ser Asn His

355 360 365

Tyr Glu Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser

370 375 380

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

385 390 395 400

Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln

405 410 415

Lys Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn

420 425 430

Tyr Val Ser Trp Tyr Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu

435 440 445

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser

450 455 460

Gly Ser Lys Ser Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln

465 470 475 480

Thr Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Ala Trp Asp Ser Ser Leu

485 490 495

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly

740

<210> 120

<211> 744

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0011-HC2-k deletion

<400> 120

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Leu Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly

740

<210> 121

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 1F

<400> 121

Phe Leu Leu Met Trp Ile Thr Gln Cys

1 5

<210> 122

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 2M

<400> 122

Ser Met Leu Met Trp Ile Thr Gln Cys

1 5

<210> 123

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 3A

<400> 123

Ser Leu Ala Met Trp Ile Thr Gln Cys

1 5

<210> 124

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 4A

<400> 124

Ser Leu Leu Ala Trp Ile Thr Gln Cys

1 5

<210> 125

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 5A

<400> 125

Ser Leu Leu Met Ala Ile Thr Gln Cys

1 5

<210> 126

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 6L

<400> 126

Ser Leu Leu Met Trp Leu Thr Gln Cys

1 5

<210> 127

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 7F

<400> 127

Ser Leu Leu Met Trp Ile Phe Gln Cys

1 5

<210> 128

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 8A

<400> 128

Ser Leu Leu Met Trp Ile Thr Ala Cys

1 5

<210> 129

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of point mutated NY-ESO peptide 9A

<400> 129

Ser Leu Leu Met Trp Ile Thr Gln Ala

1 5

<210> 130

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of gp100 peptide

<400> 130

Ile Met Asp Gln Val Pro Phe Ser Val

1 5

<210> 131

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the homologous peptide DOLPP1

<400> 131

Ser Gln Phe Met Trp Phe Phe Ser Val

1 5

<210> 132

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the IL20RB homologous peptide

<400> 132

Ser Leu Phe Met Trp Phe Phe Tyr Ala

1 5

<210> 133

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the homologous peptide PRKD2

<400> 133

Ser Leu Asp Met Trp Ser Val Gly Val

1 5

<210> 134

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the homologous peptide CD163

<400> 134

Ser Ile Pro Met Trp Val Asp Asn Val

1 5

<210> 135

<211> 9

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of the homologous peptide P2RY8

<400> 135

Ser Val Ala Met Trp Ala Val Phe Leu

1 5

<210> 136

<211> 269

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence C3E-7034

<400> 136

Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

1 5 10 15

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr

20 25 30

Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

35 40 45

Val Ala Ser Ile Thr Asn Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser

50 55 60

Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu

65 70 75 80

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro His Ser Val

130 135 140

Ser Glu Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Lys Arg Asn Thr

145 150 155 160

Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln His Glu Gly

165 170 175

Ser Ser Pro Thr Thr Ile Ile Tyr Arg Asp Asp Lys Arg Pro Asp Gly

180 185 190

Val Ser Asp Arg Phe Ser Gly Ser Ile Asp Arg Ser Ser Lys Ser Ala

195 200 205

Ser Leu Thr Ile Ser Asn Leu Lys Thr Glu Asp Glu Ala Asp Tyr Phe

210 215 220

Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Thr Val Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp

245 250 255

Asp Asp Lys Gly Ala Ala Ala His His His His His

260 265

<210> 137

<211> 267

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence C3E-7036

<400> 137

Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

1 5 10 15

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr

20 25 30

Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

35 40 45

Val Ala Ser Ile Thr Asn Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser

50 55 60

Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu

65 70 75 80

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val

130 135 140

Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr

145 150 155 160

Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly

165 170 175

Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly

180 185 190

Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu

195 200 205

Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln

210 215 220

Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val

225 230 235 240

Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Asp

245 250 255

Lys Gly Ala Ala Ala His His His His His

260 265

<210> 138

<211> 267

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence C3E-7085

<400> 138

Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

1 5 10 15

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr

20 25 30

Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

35 40 45

Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser

50 55 60

Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu

65 70 75 80

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val

130 135 140

Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr

145 150 155 160

Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly

165 170 175

Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly

180 185 190

Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu

195 200 205

Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln

210 215 220

Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val

225 230 235 240

Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Asp

245 250 255

Lys Gly Ala Ala Ala His His His His His

260 265

<210> 139

<211> 269

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence C3E-7088

<400> 139

Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

1 5 10 15

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr

20 25 30

Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

35 40 45

Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser

50 55 60

Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu

65 70 75 80

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro His Ser Val

130 135 140

Ser Glu Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Lys Arg Asn Thr

145 150 155 160

Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln His Glu Gly

165 170 175

Ser Ser Pro Thr Thr Ile Ile Tyr Arg Asn Asp Lys Arg Pro Asp Gly

180 185 190

Val Ser Asp Arg Phe Ser Gly Ser Ile Asp Arg Ser Ser Lys Ser Ala

195 200 205

Ser Leu Thr Ile Ser Asn Leu Lys Thr Glu Asp Glu Ala Asp Tyr Phe

210 215 220

Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Thr Val Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp

245 250 255

Asp Asp Lys Gly Ala Ala Ala His His His His His

260 265

<210> 140

<211> 269

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence C3E-7093

<400> 140

Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

1 5 10 15

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr

20 25 30

Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

35 40 45

Val Ala Ser Ile Thr Ser Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser

50 55 60

Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu

65 70 75 80

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro His Ser Val

130 135 140

Ser Glu Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Lys Arg Asn Thr

145 150 155 160

Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln His Glu Gly

165 170 175

Ser Ser Pro Thr Thr Ile Ile Tyr Arg Asn Asp Lys Arg Pro Asp Gly

180 185 190

Val Ser Asp Arg Phe Ser Gly Ser Ile Asp Arg Ser Ser Lys Ser Ala

195 200 205

Ser Leu Thr Ile Ser Asn Leu Lys Thr Glu Asp Glu Ala Asp Tyr Phe

210 215 220

Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Thr Val Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp

245 250 255

Asp Asp Lys Gly Ala Ala Ala His His His His His

260 265

<210> 141

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of CDR1 of the heavy chain C3E-7085

<400> 141

Gly Val Thr Phe Asn Tyr Tyr Gly

1 5

<210> 142

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of CDR2 of the heavy chain C3E-7085

<400> 142

Ile Thr Arg Ser Gly Gly Arg Ile

1 5

<210> 143

<211> 11

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of CDR3 of the heavy chain C3E-7085

<400> 143

Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr

1 5 10

<210> 144

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> C3E-7085 light chain CDR1 amino acid sequence

<400> 144

Thr Gly Asn Ile Gly Ser Asn Tyr

1 5

<210> 145

<211> 3

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of CDR2 of the light chain of C3E-7085

<400> 145

Arg Asp Asp

1

<210> 146

<211> 8

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence of CDR3 light chain C3E-7085

<400> 146

Gln Ser Tyr Ser Ser Gly Phe Ile

1 5

<210> 147

<211> 269

<212> PROTEIN

<213> Artificial

<220>

<223> Amino acid sequence C3E-7078

<400> 147

Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

1 5 10 15

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr

20 25 30

Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp

35 40 45

Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser

50 55 60

Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu

65 70 75 80

Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

115 120 125

Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro His Ser Val

130 135 140

Ser Glu Ser Pro Gly Lys Thr Val Thr Ile Ser Cys Lys Arg Asn Thr

145 150 155 160

Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln His Glu Gly

165 170 175

Ser Ser Pro Thr Thr Ile Ile Tyr Arg Asp Asp Lys Arg Pro Asp Gly

180 185 190

Val Ser Asp Arg Phe Ser Gly Ser Ile Asp Arg Ser Ser Lys Ser Ala

195 200 205

Ser Leu Thr Ile Ser Asn Leu Lys Thr Glu Asp Glu Ala Asp Tyr Phe

210 215 220

Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu

225 230 235 240

Thr Val Leu Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp

245 250 255

Asp Asp Lys Gly Ala Ala Ala His His His His His

260 265

<210> 148

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYF-0014-HC2

<400> 148

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgga 60

gaggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgacaccc 120

tcctgttcag cgtctggatt ctccatccgt agttatgata tacactgggt ccgccaggct 180

ccaggcaagg ggctagagtg ggtggccact atatcatatg atggaagtca gaagtacttc 240

gcagactccg tgaagggccg atttaccatc ttcagagacg aatcggagaa catggtgtat 300

ctgcaaatga gcggcctgag agttgaggac acggctgttt atcactgtgc gagagggagt 360

agtggtcatt atgaggcttt tgatatctgg ggccaaggga caatggtcac cgtctcttca 420

ggtggaggcg gttcaggcgg aggtggcagc ggcggtggcg ggagtcagtc tgtgttgacg 480

cagccgccct cagtgtctgc ggccccagga cagaaggtca ccatctcctg ctctggaagc 540

agctccaaca ttgggaataa ttatgtatcc tggtaccagc agctcccagg aacagccccc 600

aaactcctca tttatgacaa taataagcga ccctcaggga ttcctgaccg attctctggc 660

tccaagtctg gcacgtcagc caccctgggc atcaccggac tccagactgg ggacgaggcc 720

gattattact gcggaacatg ggatagcagc ctgagtgccc cttgggtgtt cggcggaggg 780

accaaggtca ccgtcctagg gggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

cagggaaccc tggtgacagt cagctccgga ggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac agctcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 149

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0014-HC2

<400> 149

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 150

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYF-0082-HC2

<400> 150

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Thr Pro Ser Cys Ser Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Phe

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Arg Asp Glu Ser Glu

85 90 95

Asn Met Val Tyr Leu Gln Met Ser Gly Leu Arg Val Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn Tyr Val Ser Trp Tyr

180 185 190

Gln Arg Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Ser Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 151

<211> 207

<212> PROTEIN

<213> Homo sapiens

<400> 151

Met Gln Ser Gly Thr His Trp Arg Val Leu Gly Leu Cys Leu Leu Ser

1 5 10 15

Val Gly Val Trp Gly Gln Asp Gly Asn Glu Glu Met Gly Gly Ile Thr

20 25 30

Gln Thr Pro Tyr Lys Val Ser Ile Ser Gly Thr Thr Val Ile Leu Thr

35 40 45

Cys Pro Gln Tyr Pro Gly Ser Glu Ile Leu Trp Gln His Asn Asp Lys

50 55 60

Asn Ile Gly Gly Asp Glu Asp Asp Lys Asn Ile Gly Ser Asp Glu Asp

65 70 75 80

His Leu Ser Leu Lys Glu Phe Ser Glu Leu Glu Gln Ser Gly Tyr Tyr

85 90 95

Val Cys Tyr Pro Arg Gly Ser Lys Pro Glu Asp Ala Asn Phe Tyr Leu

100 105 110

Tyr Leu Arg Ala Arg Val Cys Glu Asn Cys Met Glu Met Asp Val Met

115 120 125

Ser Val Ala Thr Ile Val Ile Val Asp Ile Cys Ile Thr Gly Gly Leu

130 135 140

Leu Leu Leu Val Tyr Tyr Trp Ser Lys Asn Arg Lys Ala Lys Ala Lys

145 150 155 160

Pro Val Thr Arg Gly Ala Gly Ala Gly Gly Arg Gln Arg Gly Gln Asn

165 170 175

Lys Glu Arg Pro Pro Pro Val Pro Asn Pro Asp Tyr Glu Pro Ile Arg

180 185 190

Lys Gly Gln Arg Asp Leu Tyr Ser Gly Leu Asn Gln Arg Arg Ile

195 200 205

<210> 152

<211> 2235

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYZ-0038-HC2

<400> 152

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg aggaggcggt tcagaagtgc agctggtgga atccgggggg 840

ggcctggtgc agcctggggg gagcctgaga ctgagttgtg ccgcctctgg ggtgacattt 900

aactactatg gcatgtcttg gatccgccag gcacctggaa agggcctgga gtgggtggcc 960

agcatcacta ggtccggcgg gcgaatctac tatcccgaca gcgtcaaggg caggttcaca 1020

atttcccgcg agaacacaca gaaaactctg tacctgcaga tgaatagcct gagagccgaa 1080

gatacagctg tgtactattg cactctggac ggcagggatg ggtgggtcgc ctattggggg 1140

tgtggaaccc tggtgacagt cagctccgga gggaggaggat ctggcggagg aggcagtggg 1200

ggaggcgggt caaactttat gctcactcag ccgtcctctg tttctggcgt acctggccaa 1260

cgggtgacca ttagctgtac gggtaatacc gggaatatcg ggtctaacta cgtgaactgg 1320

tatcagcagc ttccagggac atgtcccaag ttgctgatct atcgcgacga caaaagaccc 1380

tcaggggtcc ctgaccgatt tagtggcagc aaaagcggta cttccgcttc cctggcgata 1440

accggctttc aggccgaaga tgaggcagac tactattgcc agtcatattc cagcggcttc 1500

atcttcggag gcggaactaa gctgacagtg ttggcagccg agcccaaatc ttctgacaaa 1560

actcacacat gcccaccctg cccagcacct gaagccgcag ggggaccctc agtcttcctc 1620

ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 1680

gtggtgtcag tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 1740

gaggtgcata atgccaagac aaagccccgg gaggagcagt acaacagcac gtaccgggtg 1800

gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 1860

gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaaggccag 1920

ccccgggaac cacaggtgta cgtgtacccc ccatcccggg acgagctgac caagaaccag 1980

gtcagcctga cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 2040

agcaatggcc agcccgagaa caactacaag accacccctc ccgtgctgga ctccgacggc 2100

tccttcgccc tcgtgagcaa gctcaccgtg gacaagagca ggtggcagca gggcaacgtc 2160

ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacccagaa gagcctctcc 2220

ctgtctcccg gcaag 2235

<210> 153

<211> 2250

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYZ-0082-HC2

<400> 153

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtggagg aggtggatct 480

caatccgttc tgactcagcc accctccgcc tctggcaccc cgggccaacg ggtcacaata 540

tcatgttctg gctcttcaag caacatcgga aactggtacg tgagctggta ccagcagctc 600

cccggcacgg cgcccaagct tctgatatat gacaacaaca aacggcccag tggagttcct 660

gacagattca gtgggtctaa aagtggtaca agcgctagcc tggccataag tggtctgcag 720

agtgaagatg aggcggacta ttattgcgga acctgggact ccagcctgag cgctccctgg 780

gttttcggcg gaggtactaa gctgaccgtc ttgggaggag gcggttcaga agtgcagctg 840

gtggaatccg gggggggcct ggtgcagcct ggggggagcc tgagactgag ttgtgccgcc 900

tctggggtga catttaacta ctatggcatg tcttggatcc gccaggcacc tggaaagggc 960

ctggagtggg tggccagcat cactaggtcc ggcgggcgaa tctactatcc cgacagcgtc 1020

aagggcaggt tcacaatttc ccgcgagaac acacagaaaa ctctgtacct gcagatgaat 1080

agcctgagag ccgaagatac agctgtgtac tattgcactc tggacggcag ggatgggtgg 1140

gtcgcctatt gggggtgtgg aaccctggtg acagtcagct ccggaggagg aggatctggc 1200

ggaggaggca gtgggggagg cgggtcaaac tttatgctca ctcagccgtc ctctgtttct 1260

ggcgtacctg gccaacgggt gaccattagc tgtacgggta ataccgggaa tatcgggtct 1320

aactacgtga actggtatca gcagcttcca gggacatgtc ccaagttgct gatctatcgc 1380

gacgacaaaa gaccctcagg ggtccctgac cgatttagtg gcagcaaaag cggtacttcc 1440

gcttccctgg cgataaccgg ctttcaggcc gaagatgagg cagactacta ttgccagtca 1500

tattccagcg gcttcatctt cggaggcgga actaagctga cagtgttggc agccgagccc 1560

aaatcttctg acaaaactca cacatgccca ccctgcccag cacctgaagc cgcaggggga 1620

ccctcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 1680

gaggtcacat gcgtggtggt gtcagtgagc cacgaagacc ctgaggtcaa gttcaactgg 1740

tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cccgggagga gcagtacaac 1800

agcacgtacc gggtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 1860

gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 1920

aaagccaaag gccagccccg ggaaccacag gtgtacgtgt accccccatc ccgggacgag 1980

ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 2040

gccgtggagt gggagagcaa tggccagccc gagaacaact acaagaccac ccctcccgtg 2100

ctggactccg acggctcctt cgccctcgtg agcaagctca ccgtggacaa gagcaggtgg 2160

cagcagggca acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacc 2220

cagaagagcc tctccctgtc tcccggcaag 2250

<210> 154

<211> 2250

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYZ-0083-HC2

<400> 154

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtggagg aggtggatct 480

caatccgttc tgactcagcc accctccgcc tctggcaccc cgggccaacg ggtcacaata 540

tcatgttctg gctcttcaag caacatcgga aactggtacg tgagctggta ccagcagctc 600

cccggcacgg cgcccaagct tctgatatat gacaacaaca aacggcccag tggagttcct 660

gacagattca gtgggtctaa aagtggtaca agcgctagcc tggccataag tggtctgcag 720

agtgaagatg aggcggacta ttattgcgga acctgggact ccagcctgag cgctccctgg 780

gttttcggcg gaggtactaa gctgaccgtc ttgggaggag gcggttcaga agtgcagctg 840

gtggaatccg gggggggcct ggtgcagcct ggggggagcc tgagactgag ttgtgccgcc 900

tctggggtga catttaacta ctatggcatg tcttggatcc gccaggcacc tggaaagtgt 960

ctggagtggg tggccagcat cactaggtcc ggcgggcgaa tctactatcc cgacagcgtc 1020

aagggcaggt tcacaatttc ccgcgagaac acacagaaaa ctctgtacct gcagatgaat 1080

agcctgagag ccgaagatac agctgtgtac tattgcactc tggacggcag ggatgggtgg 1140

gtcgcctatt gggggcaggg aaccctggtg acagtcagct ccggaggagg aggatctggc 1200

ggaggaggca gtgggggagg cgggtcaaac tttatgctca ctcagccgtc ctctgtttct 1260

ggcgtacctg gccaacgggt gaccattagc tgtacgggta ataccgggaa tatcgggtct 1320

aactacgtga actggtatca gcagcttcca gggacagctc ccaagttgct gatctatcgc 1380

gacgacaaaa gaccctcagg ggtccctgac cgatttagtg gcagcaaaag cggtacttcc 1440

gcttccctgg cgataaccgg ctttcaggcc gaagatgagg cagactacta ttgccagtca 1500

tattccagcg gcttcatctt cggatgtgga actaagctga cagtgttggc agccgagccc 1560

aaatcttctg acaaaactca cacatgccca ccctgcccag cacctgaagc cgcaggggga 1620

ccctcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 1680

gaggtcacat gcgtggtggt gtcagtgagc cacgaagacc ctgaggtcaa gttcaactgg 1740

tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cccgggagga gcagtacaac 1800

agcacgtacc gggtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 1860

gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 1920

aaagccaaag gccagccccg ggaaccacag gtgtacgtgt accccccatc ccgggacgag 1980

ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 2040

gccgtggagt gggagagcaa tggccagccc gagaacaact acaagaccac ccctcccgtg 2100

ctggactccg acggctcctt cgccctcgtg agcaagctca ccgtggacaa gagcaggtgg 2160

cagcagggca acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacc 2220

cagaagagcc tctccctgtc tcccggcaag 2250

<210> 155

<211> 745

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYZ-0038-HC2

<400> 155

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Cys Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

405 410 415

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

420 425 430

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Cys

435 440 445

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

450 455 460

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

465 470 475 480

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

485 490 495

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Ala

500 505 510

Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

515 520 525

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

530 535 540

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

545 550 555 560

Val Val Ser Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

565 570 575

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

580 585 590

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

595 600 605

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

610 615 620

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

625 630 635 640

Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu

645 650 655

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

660 665 670

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

675 680 685

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu

690 695 700

Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

705 710 715 720

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

725 730 735

Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745

<210> 156

<211> 750

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYZ-0082-HC2

<400> 156

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

145 150 155 160

Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

165 170 175

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

180 185 190

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

210 215 220

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

225 230 235 240

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

245 250 255

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

260 265 270

Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

275 280 285

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr

290 295 300

Phe Asn Tyr Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly

305 310 315 320

Leu Glu Trp Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr

325 330 335

Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln

340 345 350

Lys Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

355 360 365

Val Tyr Tyr Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp

370 375 380

Gly Cys Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro

405 410 415

Ser Ser Val Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr

420 425 430

Gly Asn Thr Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln

435 440 445

Leu Pro Gly Thr Cys Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg

450 455 460

Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser

465 470 475 480

Ala Ser Leu Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr

485 490 495

Tyr Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys

500 505 510

Leu Thr Val Leu Ala Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr

515 520 525

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

530 535 540

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

545 550 555 560

Glu Val Thr Cys Val Val Val Ser Val Ser His Glu Asp Pro Glu Val

565 570 575

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

580 585 590

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

595 600 605

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

610 615 620

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

625 630 635 640

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro

645 650 655

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

660 665 670

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

675 680 685

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

690 695 700

Gly Ser Phe Ala Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

705 710 715 720

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

725 730 735

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745 750

<210> 157

<211> 750

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYZ-0083-HC2

<400> 157

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

145 150 155 160

Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

165 170 175

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

180 185 190

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

210 215 220

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

225 230 235 240

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

245 250 255

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

260 265 270

Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

275 280 285

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr

290 295 300

Phe Asn Tyr Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Cys

305 310 315 320

Leu Glu Trp Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr

325 330 335

Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln

340 345 350

Lys Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

355 360 365

Val Tyr Tyr Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp

370 375 380

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly

385 390 395 400

Gly Gly Gly Ser Gly Gly Gly Gly Ser Asn Phe Met Leu Thr Gln Pro

405 410 415

Ser Ser Val Ser Gly Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr

420 425 430

Gly Asn Thr Gly Asn Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln

435 440 445

Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg

450 455 460

Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser

465 470 475 480

Ala Ser Leu Ala Ile Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr

485 490 495

Tyr Cys Gln Ser Tyr Ser Ser Gly Phe Ile Phe Gly Cys Gly Thr Lys

500 505 510

Leu Thr Val Leu Ala Ala Glu Pro Lys Ser Ser Asp Lys Thr His Thr

515 520 525

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

530 535 540

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

545 550 555 560

Glu Val Thr Cys Val Val Val Ser Val Ser His Glu Asp Pro Glu Val

565 570 575

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

580 585 590

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

595 600 605

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

610 615 620

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

625 630 635 640

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro Pro

645 650 655

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

660 665 670

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

675 680 685

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

690 695 700

Gly Ser Phe Ala Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

705 710 715 720

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

725 730 735

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

740 745 750

<210> 158

<211> 2172

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYZ-1010-HC2

<400> 158

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtggagg aggtggatct 480

caatccgttc tgactcagcc accctccgcc tctggcaccc cgggccaacg ggtcacaata 540

tcatgttctg gctcttcaag caacatcgga aactggtacg tgagctggta ccagcagctc 600

cccggcacgg cgcccaagct tctgatatat gacaacaaca aacggcccag tggagttcct 660

gacagattca gtgggtctaa aagtggtaca agcgctagcc tggccataag tggtctgcag 720

agtgaagatg aggcggacta ttattgcgga acctgggact ccagcctgag cgctccctgg 780

gttttcggcg gaggtactaa gctgaccgtc ttgggaggag gcggttcaga agtgcagctg 840

gtggaatccg gggggggcct ggtgcagcct ggggggagcc tgagactgag ttgtgccgcc 900

tctggggtga catttaacta ctatggcatg tcttggatcc gccaggcacc tggaaagggc 960

ctggagtggg tggccagcat cactaggtcc ggcgggcgaa tctactatcc cgacagcgtc 1020

aagggcaggt tcacaatttc ccgcgagaac acacagaaaa ctctgtacct gcagatgaat 1080

agcctgagag ccgaagatac agctgtgtac tattgcactc tggacggcag ggatgggtgg 1140

gtcgcctatt ggggtcaggg aaccctggtg acagtcagct ccgcctccac caagggccca 1200

agcgtcttcc ccctggcacc ctcctccaag agcacctctg gcggcacagc cgccctgggc 1260

tgcctggtca aggactactt ccccgaaccc gtgaccgtga gctggaactc aggcgccctg 1320

accagcggcg tgcacacctt ccccgctgtc ctgcagtcct caggactcta ctccctcagc 1380

agcgtggtga ccgtgccctc cagcagcttg ggcacccaga cctacatctg caacgtgaat 1440

cacaagccca gcaacaccaa ggtggacaag agagttgagc ccaagtcttg cgacaaaact 1500

cacacatgcc caccctgccc agcacctgaa gccgcagggg gaccctcagt cttcctcttc 1560

cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg 1620

gtgtcagtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga cggcgtggag 1680

gtgcataatg ccaagacaaa gccccgggag gagcagtaca acagcacgta ccgggtggtc 1740

agcgtcctca ctgtcctgca ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc 1800

tccaacaaag ccctcccagc ccccatcgag aaaaccatct ccaaagccaa aggccagccc 1860

cgggaaccac aggtgtacgt gtacccccca tcccgggacg agctgaccaa gaaccaggtc 1920

agcctgacct gcctggtcaa aggcttctat cccagcgaca tcgccgtgga gtggggagagc 1980

aatggccagc ccgagaacaa ctacaagacc acccctcccg tgctggactc cgacggctcc 2040

ttcgccctcg tgagcaagct caccgtggac aagagcaggt ggcagcaggg caacgtcttc 2100

tcatgctccg tgatgcatga ggctctgcac aaccactaca cccagaagag cctctccctg 2160

tctcccggca ag 2172

<210> 159

<211> 699

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of C3E-7085-LC

<400> 159

atggtgctgc agacccaggt gttcatctcc ctgctgctgt ggatctccgg cgcgtacggc 60

aactttatgc tcactcagcc gtcctctgtt tctggcgtac ctggccaacg ggtgaccatt 120

agctgtacgg gtaataccgg gaatatcggg tctaactacg tgaactggta tcagcagctt 180

ccagggacag ctcccaagtt gctgatctat cgcgacgaca aaagaccctc aggggtccct 240

gaccgattta gtggcagcaa aagcggtact tccgcttccc tggcgataac cggctttcag 300

gccgaagatg aggcagacta ctattgccag tcatattcca gcggcttcat cttcggaggc 360

ggaactaagc tgacagtgct gggccagcct aaggctgccc ctagcgtgac cctgttccct 420

ccttccagcg aggagcttca agctaacaag gccaccctgg tgtgtcttat ctctgacttc 480

taccctggcg ctgtgaccgt ggcctggaag gctgacagct cccctgtgaa ggccggagtg 540

gagaccacca cacctagcaa gcagtctaac aacaagtacg ctgccagctc ctacctgagc 600

cttacccctg agcagtggaa gtctcacaga agctactcct gtcaagtgac ccacgaggc 660

agcaccgtgg agaagaccgt ggctcctacc gagtgttcc 699

<210> 160

<211> 724

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYZ-1010-HC2

<400> 160

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

145 150 155 160

Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

165 170 175

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

180 185 190

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

210 215 220

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

225 230 235 240

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

245 250 255

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

260 265 270

Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

275 280 285

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr

290 295 300

Phe Asn Tyr Tyr Gly Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly

305 310 315 320

Leu Glu Trp Val Ala Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr

325 330 335

Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln

340 345 350

Lys Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

355 360 365

Val Tyr Tyr Cys Thr Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp

370 375 380

Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

385 390 395 400

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

405 410 415

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

420 425 430

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

435 440 445

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

450 455 460

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

465 470 475 480

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

485 490 495

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

500 505 510

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

515 520 525

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ser Val Ser

530 535 540

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

545 550 555 560

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

565 570 575

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

580 585 590

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

595 600 605

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

610 615 620

Val Tyr Val Tyr Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val

625 630 635 640

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

645 650 655

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

660 665 670

Pro Val Leu Asp Ser Asp Gly Ser Phe Ala Leu Val Ser Lys Leu Thr

675 680 685

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

690 695 700

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

705 710 715 720

Ser Pro Gly Lys

<210> 161

<211> 233

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of C3E-7085-LC

<400> 161

Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser

1 5 10 15

Gly Ala Tyr Gly Asn Phe Met Leu Thr Gln Pro Ser Ser Val Ser Gly

20 25 30

Val Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Asn Thr Gly Asn

35 40 45

Ile Gly Ser Asn Tyr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala

50 55 60

Pro Lys Leu Leu Ile Tyr Arg Asp Asp Lys Arg Pro Ser Gly Val Pro

65 70 75 80

Asp Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile

85 90 95

Thr Gly Phe Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr

100 105 110

Ser Ser Gly Phe Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

115 120 125

Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu

130 135 140

Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe

145 150 155 160

Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val

165 170 175

Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys

180 185 190

Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser

195 200 205

His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu

210 215 220

Lys Thr Val Ala Pro Thr Glu Cys Ser

225 230

<210> 162

<211> 5

<212> PROTEIN

<213> Artificial

<220>

<223> Peptide linker

<400> 162

Gly Gly Gly Gly Ser

1 5

<210> 163

<211> 891

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYA-3061

<400> 163

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtggagg aggtggatct 480

caatccgttc tgactcagcc accctccgcc tctggcaccc cgggccaacg ggtcacaata 540

tcatgttctg gctcttcaag caacatcgga aactggtacg tgagctggta ccagcagctc 600

cccggcacgg cgcccaagct tctgatatat gacaacaaca aacggcccag tggagttcct 660

gacagattca gtgggtctaa aagtggtaca agcgctagcc tggccataag tggtctgcag 720

agtgaagatg aggcggacta ttattgcgga acctgggact ccagcctgag cgctccctgg 780

gttttcggcg gaggtactaa gctgaccgtc ttgggcgcgg ccgcaggtgc aggtggtgat 840

tacaaagatg atgacgataa aggtgcagcg gcgcatcacc atcatcacca c 891

<210> 164

<211> 297

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYA-3061

<400> 164

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

145 150 155 160

Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

165 170 175

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

180 185 190

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

210 215 220

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

225 230 235 240

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

245 250 255

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

260 265 270

Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly

275 280 285

Ala Ala Ala His His His His His

290 295

<210> 165

<211> 903

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0005

<400> 165

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gttgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

agctgtgtgg ccagcggctt caccttcgac gactacgcca tgtactgggt ccgacaggtg 180

ccaggcaagg acctggaatg ggtgtccctg atttctggcg acggcgacat cacctactac 240

gtggactctg tgaagggcag attcaccgtg tccagagaca acaacaagaa cagcctgtac 300

ctgcagatga agtccctgcg cgtggaagat acagccctgt actactgcgc caaggacatg 360

atctactacg cctcttggag cggctacggc agcagcgatt actactacta tgtgatggac 420

gtgtggggcc agggcaccac cgttacagtt tctagcggag gcggaggaag tggcggcgga 480

ggatctggcg gtggtggttc tgatatccag atgacacaga gccccagcag cctgtctgcc 540

tctgtgggag acagagtgac catcacctgt agagccagcc agagcatcag cagctacctg 600

aactggtatc agcagaagcc cggcaaggcc cctaaactgc tgatctatgc cgcctccagt 660

ctgcagagcg gagtgcctag cagattttct ggcagcggct ccggcaccga tttcaccctg 720

accatatcta gcctgcagcc tgaggacttc gccacctact attgccagca gagctacagc 780

acccctccta tcacctttgg ccagggaacc agactggaaa tcaaaggcgc tgctgcaggc 840

gctggcggcg actacaaaga cgatgatgat aagggcgctg ccgctcacca cccacatcac 900

cat 903

<210> 166

<211> 301

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0005

<400> 166

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met Tyr Trp Val Arg Gln Val Pro Gly Lys Asp

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Asp Ile Thr Tyr Tyr

65 70 75 80

Val Asp Ser Val Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Asn Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Lys Ser Leu Arg Val Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Ile Tyr Tyr Ala Ser Trp Ser Gly

115 120 125

Tyr Gly Ser Ser Asp Tyr Tyr Tyr Tyr Val Met Asp Val Trp Gly Gln

130 135 140

Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

165 170 175

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

180 185 190

Ser Gln Ser Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

195 200 205

Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly

210 215 220

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

225 230 235 240

Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln

245 250 255

Gln Ser Tyr Ser Thr Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu

260 265 270

Glu Ile Lys Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp

275 280 285

Asp Asp Lys Gly Ala Ala Ala His His His His His

290 295 300

<210> 167

<211> 903

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0006

<400> 167

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gttgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

tcttgtgccg ccagcggctt caccttcgac gactacgcca tgtactgggt ccgacaggcc 180

cctggcaaag gccttgaatg ggtgtccctg atcagcggag atggcggcag catgtactac 240

gccgatagcg tgaagggcag attcaccatc agccgggaca acagcaagaa cagcctgtac 300

ctgcagatga actccctgcg gaccgaggac acagccctgt actactgcgc caaggatatg 360

atcttctacg ccttttggag cggctacggc agcagcgatt actactacta cgtgatggac 420

gtgtggggcc agggcaccac agtgacagtt tattctggcg gcggaggatc tggcggaggt 480

ggaagcggag gcggtggatc tgatatccag atgacacaga gccccagcag cctgtctgcc 540

tctgtgggag acagagtgac catcacctgt cgggccagcc agagaatcag cacctacctg 600

aactggtatc agcagaagcc cggcaaggcc cctaagctgc tgatctatgc tgcctccagt 660

ctgcagagcg gcgtgccaag cagattttct ggcagcggct ctggcaccga cttcaccctg 720

accatatcta gcctgcagcc tgaggacttc gccacctact actgccagca gagctacagc 780

acccctccta tcacctttgg ccagggaacc agactggaaa tcaaaggcgc tgctgcaggc 840

gcaggcggcg actacaaaga cgatgatgat aagggcgctg ccgctcacca cccacatcac 900

cat 903

<210> 168

<211> 301

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0006

<400> 168

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Gly Ser Met Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Ile Phe Tyr Ala Phe Trp Ser Gly

115 120 125

Tyr Gly Ser Ser Asp Tyr Tyr Tyr Tyr Val Met Asp Val Trp Gly Gln

130 135 140

Gly Thr Thr Val Thr Val Tyr Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

165 170 175

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

180 185 190

Ser Gln Arg Ile Ser Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

195 200 205

Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly

210 215 220

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

225 230 235 240

Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln

245 250 255

Gln Ser Tyr Ser Thr Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu

260 265 270

Glu Ile Lys Gly Ala Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp

275 280 285

Asp Asp Lys Gly Ala Ala Ala His His His His His

290 295 300

<210> 169

<211> 888

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0007

<400> 169

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gttgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

tcttgtgccg ccagcggctt caccttcgac gattatgcca tgcactgggt ccgacaggcc 180

cctggcaaag gacttgaatg ggtgtccctg atcagcggcg acggcgacaa tacctactac 240

gccgatagcg tgaagggcag attcaccatc agccgggaca acaacaagaa cagcctgtac 300

ctgcagatga actccctgcg gaccgaggac accgccttct actactgtgc caaagagctg 360

atcttcggca aggtgctgca cgacttttac tactacgtga tggacgtgtg gggccagggc 420

acccacagtga cagtttctag cggaggcgga ggaagtggcg gcggaggatc tggcggtggt 480

ggttctgata tccagatgac acagagcccc agcagcctgt ctgcctctgt ggggacaga 540

gtgaccatca cctgtagagc cagccagagc atcagcagct acctgaactg gtatcagcag 600

aagcccggca aggcccctaa actgctgatc tatgccgcct ccagtctgca gagcggagtg 660

cctagcagat tttctggcag cggctccggc accgatttca ccctgaccat atctagcctg 720

cagcctgagg acttcgccac ctactactgc cagcagagct acagcacccc tcctatcacc 780

tttggccagg gaaccagact ggaaatcaaa ggcgctgctg caggcgcagg cggcgactac 840

aaagacgatg atgataaggg cgctgccgct caccaccacc atcaccat 888

<210> 170

<211> 296

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0007

<400> 170

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Asp Asn Thr Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Asn Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala

100 105 110

Phe Tyr Tyr Cys Ala Lys Glu Leu Ile Phe Gly Lys Val Leu His Asp

115 120 125

Phe Tyr Tyr Tyr Val Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr

130 135 140

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

165 170 175

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

180 185 190

Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

195 200 205

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

225 230 235 240

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

245 250 255

Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Ala

260 265 270

Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Lys Gly Ala

275 280 285

Ala Ala His His His His His

290 295

<210> 171

<211> 888

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0008

<400> 171

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gttgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

agctgtgaag cctccggctt catcttcgac gactacgcca tgcactgggt ccgacaggca 180

cctggcaaag gccttgaatg ggtgtccctg atctctggcg acggcgacat catctactac 240

gccgactctg tgaagggcag attcaccatc agccgggaca acagcaagaa cagcctgtac 300

ctgcagatga actccctgat catcgaggac acagccctgt actactgcgc caaggattgg 360

gtgttcggcg tcgtgatgac ccactactgg tacttcggcc tggatgtgtg gggccaggga 420

acccacagtga cagtttctag cggaggcgga ggaagtggcg gcggaggatc tggcggtggt 480

ggttctgata tccagatgac acagagcccc agcagcctgt ctgcctctga aggcgacaga 540

gtgaccatca cctgtagagc cagccagagc atcagcacct acctgaactg gtatcagcag 600

aagcccggca aggcccctaa gctgctgatc tatggtgcct ccagtctgca gagcggcgtg 660

ccaagcagat tttctggcag cggctctggc accgacttca ccctgaccat atctagcctg 720

cagcctgagg acttcgccac ctactactgc cagcagtcct acagcacccc tcctatcaca 780

tttggccagg gcaccaaggt ggaaatcaaa ggcgctgctg caggcgctgg cggcgactac 840

aaagacgatg atgataaggg cgctgccgct caccaccacc atcaccat 888

<210> 172

<211> 296

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0008

<400> 172

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Ile

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Asp Ile Ile Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Ile Ile Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Trp Val Phe Gly Val Val Met Thr His

115 120 125

Tyr Trp Tyr Phe Gly Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr

130 135 140

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

165 170 175

Glu Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

180 185 190

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

195 200 205

Leu Ile Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

225 230 235 240

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

245 250 255

Pro Pro Ile Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Ala

260 265 270

Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Lys Gly Ala

275 280 285

Ala Ala His His His His His

290 295

<210> 173

<211> 885

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0009

<400> 173

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gttgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

tcttgtgccg ccagcggctt caccttcgac gattatgcca tgcactgggt ccgacaggcc 180

cctggcaaag gacttgaatg ggtgtccctg atctctggcg gaggcggcgg aacctactac 240

agcgattctg tgaagggcag attcaccatc agccgggaca ccagcaagga cagcctgtac 300

ctgcagatga acagcctgcg gaccgaggac acagccctgt actactgcgc caaggacatg 360

gtgttcggcg tggtcacccc ttactactac ttcgccctgg atgtgtgggg ccagggcaca 420

acagtgacag tctcttctgg cggcggagga agcggaggcg gaggatccgg tggtggtgga 480

tctgacatcc agatgacaca gagccccagc agcctgtctg cctctgtggg agacagagtg 540

accatcacct gtagagccag ccagagcatc aacagctacc tgaactggta tcagcagaag 600

cccggcaagg cccctaagct gctgatctat gctgcctcca gtctgcagag cggcgtgcca 660

agcagatttt ctggcagcgg ctctggcacc gacttcaccc tgaccatatc tagcctgcag 720

cctgaggact tcgccaccta ctactgccag cagagctaca gcgcccctcc tatcacattt 780

ggccagggaa ccagactgga aatcaaaggc gctgctgcag gcgcaggcgg cgactacaaa 840

gacgatgatg ataagggcgc tgccgctcac caccatcacc atcat 885

<210> 174

<211> 295

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0009

<400> 174

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Gly Gly Gly Gly Thr Tyr Tyr

65 70 75 80

Ser Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys

85 90 95

Asp Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Val Phe Gly Val Val Thr Pro Tyr

115 120 125

Tyr Tyr Phe Ala Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val

130 135 140

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val

165 170 175

Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Ser

180 185 190

Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser

210 215 220

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln

225 230 235 240

Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Ala Pro

245 250 255

Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Ala Ala

260 265 270

Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala

275 280 285

Ala His His His His His

290 295

<210> 175

<211> 888

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0010

<400> 175

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gttgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

agctgtgctg tgtccggctt caccttcgac gactacgcca tgcactgggt ccgacaggca 180

cctggcaaag gccttgaatg ggtgtccctg atcagcggag atggcggcag cacacactac 240

gccgattctg tgaagggcag attcaccatc agccgggaca acagcaagaa cagcctgtac 300

ctgcagatga actccctgcg gacaggcgac acagccctgt actactgcgc caaggacatg 360

atcttcgccg tggtcatcac cgactaccac tactacggca tggacgtgtg gggccaggga 420

acccacagtga cagtttctag cggaggcgga ggaagtggcg gcggaggatc tggcggtggt 480

ggttctgata tccagatgac acagagcccc agcagcctgt ctgcctctgt ggggacaga 540

gtgaccatca cctgtagagc cagccagagc atcagcagct acctgaactg gtatcagcag 600

aagcccggca aggcccctaa actgctgatc tatgccgcct ccagtctgca gagcggagtg 660

cctagcagat tttctggcag cggctccggc accgatttca ccctgaccat atctagcctg 720

cagcctgagg acttcgccac ctactactgc cagcagagct acagcacccc tcctatcacc 780

tttggccagg gcaccagact ggaaatcaaa ggcgctgctg caggcgctgg cggcgactac 840

aaagacgatg atgataaggg cgctgccgct caccaccacc atcaccat 888

<210> 176

<211> 296

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0010

<400> 176

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Gly Ser Thr His Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Gly Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Ile Phe Ala Val Val Ile Thr Asp

115 120 125

Tyr His Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr

130 135 140

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

165 170 175

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

180 185 190

Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

195 200 205

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

225 230 235 240

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

245 250 255

Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Ala

260 265 270

Ala Ala Gly Ala Gly Gly Asp Tyr Lys Asp Asp Asp Lys Gly Ala

275 280 285

Ala Ala His His His His His

290 295

<210> 177

<211> 738

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of HC-h

<400> 177

atgaaacacc tgtggttctt cctcctgctg gtggcagctc ccagatgggt gctgagcgac 60

aaaactcaca catgcccacc ctgcccagca cctgaagccg ccggcggacc ctcagtcttc 120

ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 180

gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 240

gtggaggtgc ataatgccaa gaccaagcct agagaggaac agtacaacag cacctaccgg 300

gtggtgtccg tgctgacagt gctgcaccag gactggctga acggcaaaga gtacaagtgc 360

aaggtgtcca acaaggccct gcctgccccc atcgagaaaa ccatcagcaa ggccaagggc 420

cagccccgcg aacctcaagt gtgcaccctg ccaccctccc gggatgagct gaccaagaac 480

caggtgtccc tgagctgtgc cgtgaagggc ttctacccct ccgatatcgc cgtggaatgg 540

gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggacagcgac 600

ggctcattct tcctggtgtc caagctgacc gtggacaagt ccagatggca gcagggcaac 660

gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacaccca gaagagcctc 720

tccctgtctc ccggcaaa 738

<210> 178

<211> 246

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of HC-h

<400> 178

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

20 25 30

Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

35 40 45

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

50 55 60

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

65 70 75 80

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

85 90 95

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

100 105 110

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

115 120 125

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

130 135 140

Pro Gln Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn

145 150 155 160

Gln Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile

165 170 175

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

180 185 190

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys

195 200 205

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

210 215 220

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

225 230 235 240

Ser Leu Ser Pro Gly Lys

245

<210> 179

<211> 1500

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYD-2047-HC-k

<400> 179

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggt 60

caagtacagc tggtggaatc cggtggaggc gtggtccagc cgggacgcag cttgagactg 120

tcctgcgctg catctggctt ttccatacga tcctacgata tgcactgggt tcgccaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattac 240

gcggattcag tgaaggggcg gttcacaatt tcacgggaca attcaaagaa taccttgtat 300

ctccagatgt cttcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgtgagtgc agcgcctgga cagaaggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggaa tccctgacag attcagtggg 660

tctaaaagtg gtacaagcgc taccctgggt atcaccggat tgcagaccgg agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaaggtta ccgtcttggg cggagaaccc aagagcgcag acaagaccca cacctgtcct 840

ccatgtcctg ctccagaagc tgcaggcggc ccttccgtgt ttctgttccc tccaaagcct 900

aaggacaccc tgatgatcag ccggacacct gaagtgacct gcgtggtggt ggatgtgtcc 960

cacgaggatc ccgaagtgaa gttcaattgg tacgtggacg gcgtggaagt gcacaacgcc 1020

aagaccaagc ctagagagga acagtacaac agcacctaca gagtggtgtc tgtgctgaca 1080

gtgctgcacc aggactggct gaacggcaaa gagtacaagt gcaaggtgtc caacaaggcc 1140

ctgcctgctc ctatcgagaa aaccatcagc aaggccaagg gccagcctag ggaaccccag 1200

gtttacaccc tgcctccatg cagggatgag ctgaccaaga accaggtgtc cctgtggtgc 1260

ctggttaagg gcttctaccc ctccgatatc gccgtggaat ggggagagcaa tggccagcca 1320

gagaacaact acaagacaac ccctcctgtg ctggactccg acggctcatt cttcctgtac 1380

agcaagctga ccgtggacaa gagcagatgg cagcagggca acgtgttcag ctgcagcgtg 1440

atgcacgagg ccctgcacaa cactacaca cagaagtccc tgtctctgag ccccggcaaa 1500

<210> 180

<211> 500

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYD-2047-HC-k

<400> 180

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Glu Pro Lys Ser

260 265 270

Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

275 280 285

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

290 295 300

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

305 310 315 320

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

325 330 335

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

340 345 350

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

355 360 365

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

370 375 380

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

385 390 395 400

Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val

405 410 415

Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

420 425 430

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

435 440 445

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

450 455 460

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

465 470 475 480

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

485 490 495

Ser Pro Gly Lys

500

<210> 181

<211> 1500

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYD-2061-HC-k

<400> 181

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtcaatc cgttctgact 480

cagccaccct ccgcctctgg caccccgggc caacgggtca caatatcatg ttctggctct 540

tcaagcaaca tcggaaactg gtacgtgagc tggtaccagc agctccccgg cacggcgccc 600

aagcttctga tatatgacaa caacaaacgg cccagtggag ttcctgacag attcagtggg 660

tctaaaagtg gtacaagcgc tagcctggcc ataagtggtc tgcagagtga agatgaggcg 720

gactattatt gcggaacctg ggactccagc ctgagcgctc cctgggtttt cggcggaggt 780

actaagctga ccgtcttggg cggagaaccc aagagcgcag acaagaccca cacctgtcct 840

ccatgtcctg ctccagaagc tgcaggcggc ccttccgtgt ttctgttccc tccaaagcct 900

aaggacaccc tgatgatcag ccggacacct gaagtgacct gcgtggtggt ggatgtgtcc 960

cacgaggatc ccgaagtgaa gttcaattgg tacgtggacg gcgtggaagt gcacaacgcc 1020

aagaccaagc ctagagagga acagtacaac agcacctaca gagtggtgtc tgtgctgaca 1080

gtgctgcacc aggactggct gaacggcaaa gagtacaagt gcaaggtgtc caacaaggcc 1140

ctgcctgctc ctatcgagaa aaccatcagc aaggccaagg gccagcctag ggaaccccag 1200

gtttacaccc tgcctccatg cagggatgag ctgaccaaga accaggtgtc cctgtggtgc 1260

ctggttaagg gcttctaccc ctccgatatc gccgtggaat ggggagagcaa tggccagcca 1320

gagaacaact acaagacaac ccctcctgtg ctggactccg acggctcatt cttcctgtac 1380

agcaagctga ccgtggacaa gagcagatgg cagcagggca acgtgttcag ctgcagcgtg 1440

atgcacgagg ccctgcacaa cactacaca cagaagtccc tgtctctgag ccccggcaaa 1500

<210> 182

<211> 500

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYD-2061-HC-k

<400> 182

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Glu Pro Lys Ser

260 265 270

Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala

275 280 285

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

290 295 300

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

305 310 315 320

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

325 330 335

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

340 345 350

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

355 360 365

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

370 375 380

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

385 390 395 400

Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val

405 410 415

Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

420 425 430

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

435 440 445

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

450 455 460

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

465 470 475 480

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

485 490 495

Ser Pro Gly Lys

500

<210> 183

<211> 1515

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYD-3061-HC-k

<400> 183

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggt 60

gaggtacagc tggtggaatc cggtggaggc cttgtgcaac cgggaggctc cctgcgactc 120

tcttgcgccg catctggctt ttccatacga tcctacgata ttcattgggt ccgacaagcc 180

cctggcaaag gtcttgaatg ggttgccact atctcctacg acgggtctca gaaatattat 240

gcggattcag tgaaggggcg gttcacaatt tcacgagacg agtcaaagaa tacactgtac 300

ctccaaatga attcactgag agccgaggat actgcagtct atcattgtgc aagagggtcc 360

tcaggccact acgaggcctt tgatatatgg ggccaaggca ccttggtaac cgttagtagc 420

ggaggtggag gaagcggagg cggcggttcc ggaggaggtg gaagtggagg aggtggatct 480

caatccgttc tgactcagcc accctccgcc tctggcaccc cgggccaacg ggtcacaata 540

tcatgttctg gctcttcaag caacatcgga aactggtacg tgagctggta ccagcagctc 600

cccggcacgg cgcccaagct tctgatatat gacaacaaca aacggcccag tggagttcct 660

gacagattca gtgggtctaa aagtggtaca agcgctagcc tggccataag tggtctgcag 720

agtgaagatg aggcggacta ttattgcgga acctgggact ccagcctgag cgctccctgg 780

gttttcggcg gaggtactaa gctgaccgtc ttgggcggag aacccaagag cgcagacaag 840

acccacacct gtcctccatg tcctgctcca gaagctgcag gcggcccttc cgtgtttctg 900

ttccctccaa agcctaagga caccctgatg atcagccgga cacctgaagt gacctgcgtg 960

gtggtggatg tgtcccacga ggatcccgaa gtgaagttca attggtacgt ggacggcgtg 1020

gaagtgcaca acgccaagac caagcctaga gaggaacagt acaacagcac ctacagagtg 1080

gtgtctgtgc tgacagtgct gcaccaggac tggctgaacg gcaaagagta caagtgcaag 1140

gtgtccaaca aggccctgcc tgctcctatc gagaaaacca tcagcaaggc caagggccag 1200

cctagggaac cccaggttta caccctgcct ccatgcaggg atgagctgac caagaaccag 1260

gtgtccctgt ggtgcctggt taagggcttc tacccctccg atatcgccgt ggaatgggag 1320

agcaatggcc agccagagaa caactacaag acaacccctc ctgtgctgga ctccgacggc 1380

tcattcttcc tgtacagcaa gctgaccgtg gacaagagca gatggcagca gggcaacgtg 1440

ttcagctgca gcgtgatgca cgaggccctg cacaaccact acacacagaa gtccctgtct 1500

ctgagccccg gcaaa 1515

<210> 184

<211> 505

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYD-3061-HC-k

<400> 184

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

145 150 155 160

Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

165 170 175

Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp

180 185 190

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Asp Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

210 215 220

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

225 230 235 240

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu

245 250 255

Ser Ala Pro Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly

260 265 270

Gly Glu Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro

275 280 285

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

290 295 300

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

305 310 315 320

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

325 330 335

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

340 345 350

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

355 360 365

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

370 375 380

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

385 390 395 400

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu

405 410 415

Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro

420 425 430

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

435 440 445

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

450 455 460

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

465 470 475 480

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

485 490 495

Lys Ser Leu Ser Leu Ser Pro Gly Lys

500 505

<210> 185

<211> 1527

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0011-HC-k

<400> 185

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

agctgtgtgg ccagcggctt caccttcgac gactacgcca tgtactgggt ccgacaggtg 180

ccaggcaagg acctggaatg ggtgtccctg atttctggcg acggcgacat cacctactac 240

gtggactctg tgaagggcag attcaccgtg tccagagaca acaacaagaa cagcctgtac 300

ctgcagatga agtccctgcg cgtggaagat acagccctgt actactgcgc caaggacatg 360

atctactacg cctcttggag cggctacggc agcagcgatt actactacta tgtgatggac 420

gtgtggggcc agggcaccac cgttacagtt tctagcggag gcggaggaag tggcggcgga 480

ggatctggcg gtggtggttc tgatatccag atgacacaga gccccagcag cctgtctgcc 540

tctgtgggag acagagtgac catcacctgt agagccagcc agagcatcag cagctacctg 600

aactggtatc agcagaagcc cggcaaggcc cctaaactgc tgatctatgc cgcctccagt 660

ctgcagagcg gagtgcctag cagattttct ggcagcggct ccggcaccga tttcaccctg 720

accatatcta gcctgcagcc tgaggacttc gccacctact attgccagca gagctacagc 780

acccctccta tcacctttgg ccagggaacc agactggaaa tcaaaggcgg agaacccaag 840

agcgcagaca agacccacac ctgtcctcca tgtcctgctc cagaagctgc aggcggccct 900

tccgtgtttc tgttccctcc aaagcctaag gacaccctga tgatcagccg gacacctgaa 960

gtgacctgcg tggtggtgga tgtgtcccac gaggatcccg aagtgaagtt caattggtac 1020

gtggacggcg tggaagtgca caacgccaag accaagccta gagaggaaca gtacaacagc 1080

acctacagag tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaagag 1140

tacaagtgca aggtgtccaa caaggccctg cctgctccta tcgagaaaac catcagcaag 1200

gccaagggcc agcctaggga accccaggtt tacaccctgc ctccatgcag ggatgagctg 1260

accaagaacc aggtgtccct gtggtgcctg gttaagggct tctacccctc cgatatcgcc 1320

gtggaatggg agagcaatgg ccagccagag aacaactaca agacaacccc tcctgtgctg 1380

gactccgacg gctcattctt cctgtacagc aagctgaccg tggacaagag cagatggcag 1440

cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacacag 1500

aagtccctgt ctctgagccc cggcaaa 1527

<210> 186

<211> 509

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0011-HC-k

<400> 186

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met Tyr Trp Val Arg Gln Val Pro Gly Lys Asp

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Asp Ile Thr Tyr Tyr

65 70 75 80

Val Asp Ser Val Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Asn Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Lys Ser Leu Arg Val Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Ile Tyr Tyr Ala Ser Trp Ser Gly

115 120 125

Tyr Gly Ser Ser Asp Tyr Tyr Tyr Tyr Val Met Asp Val Trp Gly Gln

130 135 140

Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

165 170 175

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

180 185 190

Ser Gln Ser Ile Ser Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

195 200 205

Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly

210 215 220

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

225 230 235 240

Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln

245 250 255

Gln Ser Tyr Ser Thr Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu

260 265 270

Glu Ile Lys Gly Gly Glu Pro Lys Ser Ala Asp Lys Thr His Thr Cys

275 280 285

Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu

290 295 300

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu

305 310 315 320

Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys

325 330 335

Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys

340 345 350

Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu

355 360 365

Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys

370 375 380

Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys

385 390 395 400

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys

405 410 415

Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys

420 425 430

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln

435 440 445

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly

450 455 460

Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln

465 470 475 480

Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn

485 490 495

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

500 505

<210> 187

<211> 1527

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0012-HC-k

<400> 187

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

tcttgtgccg ccagcggctt caccttcgac gactacgcca tgtactgggt ccgacaggcc 180

cctggcaaag gccttgaatg ggtgtccctg atcagcggag atggcggcag catgtactac 240

gccgatagcg tgaagggcag attcaccatc agccgggaca acagcaagaa cagcctgtac 300

ctgcagatga actccctgcg gaccgaggac acagccctgt actactgcgc caaggatatg 360

atcttctacg ccttttggag cggctacggc agcagcgatt actactacta cgtgatggac 420

gtgtggggcc agggcaccac agtgacagtt tattctggcg gcggaggatc tggcggaggt 480

ggaagcggag gcggtggatc tgatatccag atgacacaga gccccagcag cctgtctgcc 540

tctgtgggag acagagtgac catcacctgt cgggccagcc agagaatcag cacctacctg 600

aactggtatc agcagaagcc cggcaaggcc cctaagctgc tgatctatgc tgcctccagt 660

ctgcagagcg gcgtgccaag cagattttct ggcagcggct ctggcaccga cttcaccctg 720

accatatcta gcctgcagcc tgaggacttc gccacctact actgccagca gagctacagc 780

acccctccta tcacctttgg ccagggaacc agactggaaa tcaaaggcgg agaacccaag 840

agcgcagaca agacccacac ctgtcctcca tgtcctgctc cagaagctgc aggcggccct 900

tccgtgtttc tgttccctcc aaagcctaag gacaccctga tgatcagccg gacacctgaa 960

gtgacctgcg tggtggtgga tgtgtcccac gaggatcccg aagtgaagtt caattggtac 1020

gtggacggcg tggaagtgca caacgccaag accaagccta gagaggaaca gtacaacagc 1080

acctacagag tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaagag 1140

tacaagtgca aggtgtccaa caaggccctg cctgctccta tcgagaaaac catcagcaag 1200

gccaagggcc agcctaggga accccaggtt tacaccctgc ctccatgcag ggatgagctg 1260

accaagaacc aggtgtccct gtggtgcctg gttaagggct tctacccctc cgatatcgcc 1320

gtggaatggg agagcaatgg ccagccagag aacaactaca agacaacccc tcctgtgctg 1380

gactccgacg gctcattctt cctgtacagc aagctgaccg tggacaagag cagatggcag 1440

cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacacag 1500

aagtccctgt ctctgagccc cggcaaa 1527

<210> 188

<211> 509

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0012-HC-k

<400> 188

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Gly Ser Met Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Ile Phe Tyr Ala Phe Trp Ser Gly

115 120 125

Tyr Gly Ser Ser Asp Tyr Tyr Tyr Tyr Val Met Asp Val Trp Gly Gln

130 135 140

Gly Thr Thr Val Thr Val Tyr Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser

165 170 175

Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala

180 185 190

Ser Gln Arg Ile Ser Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly

195 200 205

Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly

210 215 220

Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu

225 230 235 240

Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln

245 250 255

Gln Ser Tyr Ser Thr Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu

260 265 270

Glu Ile Lys Gly Gly Glu Pro Lys Ser Ala Asp Lys Thr His Thr Cys

275 280 285

Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu

290 295 300

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu

305 310 315 320

Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys

325 330 335

Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys

340 345 350

Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu

355 360 365

Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys

370 375 380

Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys

385 390 395 400

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys

405 410 415

Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys

420 425 430

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln

435 440 445

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly

450 455 460

Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln

465 470 475 480

Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn

485 490 495

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

500 505

<210> 189

<211> 1512

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0013-HC-k

<400> 189

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

tcttgtgccg ccagcggctt caccttcgac gattatgcca tgcactgggt ccgacaggcc 180

cctggcaaag gacttgaatg ggtgtccctg atcagcggcg acggcgacaa tacctactac 240

gccgatagcg tgaagggcag attcaccatc agccgggaca acaacaagaa cagcctgtac 300

ctgcagatga actccctgcg gaccgaggac accgccttct actactgtgc caaagagctg 360

atcttcggca aggtgctgca cgacttttac tactacgtga tggacgtgtg gggccagggc 420

acccacagtga cagtttctag cggaggcgga ggaagtggcg gcggaggatc tggcggtggt 480

ggttctgata tccagatgac acagagcccc agcagcctgt ctgcctctgt ggggacaga 540

gtgaccatca cctgtagagc cagccagagc atcagcagct acctgaactg gtatcagcag 600

aagcccggca aggcccctaa actgctgatc tatgccgcct ccagtctgca gagcggagtg 660

cctagcagat tttctggcag cggctccggc accgatttca ccctgaccat atctagcctg 720

cagcctgagg acttcgccac ctactactgc cagcagagct acagcacccc tcctatcacc 780

tttggccagg gaaccagact ggaaatcaaa ggcggagaac ccaagagcgc agacaagacc 840

cacacctgtc ctccatgtcc tgctccagaa gctgcaggcg gcccttccgt gtttctgttc 900

cctccaaagc ctaaggacac cctgatgatc agccggacac ctgaagtgac ctgcgtggtg 960

gtggatgtgt cccacgagga tcccgaagtg aagttcaatt ggtacgtgga cggcgtggaa 1020

gtgcacaacg ccaagaccaa gcctagagag gaacagtaca acagcaccta cagagtggtg 1080

tctgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140

tccaacaagg ccctgcctgc tcctatcgag aaaaccatca gcaaggccaa gggccagcct 1200

agggaacccc aggtttacac cctgcctcca tgcagggatg agctgaccaa gaaccaggtg 1260

tccctgtggt gcctggttaa gggcttctac ccctccgata tcgccgtgga atggggagagc 1320

aatggccagc cagagaacaa ctacaagaca acccctcctg tgctggactc cgacggctca 1380

ttcttcctgt acagcaagct gaccgtggac aagagcagat ggcagcaggg caacgtgttc 1440

agctgcagcg tgatgcacga ggccctgcac aaccactaca cacagaagtc cctgtctctg 1500

agccccggca aa 1512

<210> 190

<211> 504

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0013-HC-k

<400> 190

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Asp Asn Thr Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Asn Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala

100 105 110

Phe Tyr Tyr Cys Ala Lys Glu Leu Ile Phe Gly Lys Val Leu His Asp

115 120 125

Phe Tyr Tyr Tyr Val Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr

130 135 140

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

165 170 175

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

180 185 190

Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

195 200 205

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

225 230 235 240

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

245 250 255

Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Gly

260 265 270

Glu Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

275 280 285

Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

290 295 300

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

305 310 315 320

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

325 330 335

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

340 345 350

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

355 360 365

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

370 375 380

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

385 390 395 400

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr

405 410 415

Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser

420 425 430

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

435 440 445

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

450 455 460

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

465 470 475 480

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

485 490 495

Ser Leu Ser Leu Ser Pro Gly Lys

500

<210> 191

<211> 1512

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0014-HC-k

<400> 191

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

agctgtgaag cctccggctt catcttcgac gactacgcca tgcactgggt ccgacaggca 180

cctggcaaag gccttgaatg ggtgtccctg atctctggcg acggcgacat catctactac 240

gccgactctg tgaagggcag attcaccatc agccgggaca acagcaagaa cagcctgtac 300

ctgcagatga actccctgat catcgaggac acagccctgt actactgcgc caaggattgg 360

gtgttcggcg tcgtgatgac ccactactgg tacttcggcc tggatgtgtg gggccaggga 420

acccacagtga cagtttctag cggaggcgga ggaagtggcg gcggaggatc tggcggtggt 480

ggttctgata tccagatgac acagagcccc agcagcctgt ctgcctctga aggcgacaga 540

gtgaccatca cctgtagagc cagccagagc atcagcacct acctgaactg gtatcagcag 600

aagcccggca aggcccctaa gctgctgatc tatggtgcct ccagtctgca gagcggcgtg 660

ccaagcagat tttctggcag cggctctggc accgacttca ccctgaccat atctagcctg 720

cagcctgagg acttcgccac ctactactgc cagcagtcct acagcacccc tcctatcaca 780

tttggccagg gcaccaaggt ggaaatcaaa ggcggagaac ccaagagcgc agacaagacc 840

cacacctgtc ctccatgtcc tgctccagaa gctgcaggcg gcccttccgt gtttctgttc 900

cctccaaagc ctaaggacac cctgatgatc agccggacac ctgaagtgac ctgcgtggtg 960

gtggatgtgt cccacgagga tcccgaagtg aagttcaatt ggtacgtgga cggcgtggaa 1020

gtgcacaacg ccaagaccaa gcctagagag gaacagtaca acagcaccta cagagtggtg 1080

tctgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140

tccaacaagg ccctgcctgc tcctatcgag aaaaccatca gcaaggccaa gggccagcct 1200

agggaacccc aggtttacac cctgcctcca tgcagggatg agctgaccaa gaaccaggtg 1260

tccctgtggt gcctggttaa gggcttctac ccctccgata tcgccgtgga atggggagagc 1320

aatggccagc cagagaacaa ctacaagaca acccctcctg tgctggactc cgacggctca 1380

ttcttcctgt acagcaagct gaccgtggac aagagcagat ggcagcaggg caacgtgttc 1440

agctgcagcg tgatgcacga ggccctgcac aaccactaca cacagaagtc cctgtctctg 1500

agccccggca aa 1512

<210> 192

<211> 504

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0014-HC-k

<400> 192

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Ile

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Asp Ile Ile Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Ile Ile Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Trp Val Phe Gly Val Val Met Thr His

115 120 125

Tyr Trp Tyr Phe Gly Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr

130 135 140

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

165 170 175

Glu Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

180 185 190

Thr Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

195 200 205

Leu Ile Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

225 230 235 240

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

245 250 255

Pro Pro Ile Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Gly

260 265 270

Glu Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

275 280 285

Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

290 295 300

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

305 310 315 320

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

325 330 335

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

340 345 350

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

355 360 365

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

370 375 380

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

385 390 395 400

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr

405 410 415

Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser

420 425 430

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

435 440 445

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

450 455 460

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

465 470 475 480

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

485 490 495

Ser Leu Ser Leu Ser Pro Gly Lys

500

<210> 193

<211> 1509

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0015-HC-k

<400> 193

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

tcttgtgccg ccagcggctt caccttcgac gattatgcca tgcactgggt ccgacaggcc 180

cctggcaaag gacttgaatg ggtgtccctg atctctggcg gaggcggcgg aacctactac 240

agcgattctg tgaagggcag attcaccatc agccgggaca ccagcaagga cagcctgtac 300

ctgcagatga acagcctgcg gaccgaggac acagccctgt actactgcgc caaggacatg 360

gtgttcggcg tggtcacccc ttactactac ttcgccctgg atgtgtgggg ccagggcaca 420

acagtgacag tctcttctgg cggcggagga agcggaggcg gaggatccgg tggtggtgga 480

tctgacatcc agatgacaca gagccccagc agcctgtctg cctctgtggg agacagagtg 540

accatcacct gtagagccag ccagagcatc aacagctacc tgaactggta tcagcagaag 600

cccggcaagg cccctaagct gctgatctat gctgcctcca gtctgcagag cggcgtgcca 660

agcagatttt ctggcagcgg ctctggcacc gacttcaccc tgaccatatc tagcctgcag 720

cctgaggact tcgccaccta ctactgccag cagagctaca gcgcccctcc tatcacattt 780

ggccagggaa ccagactgga aatcaaaggc ggagaaccca agagcgcaga caagacccac 840

acctgtcctc catgtcctgc tccagaagct gcaggcggcc cttccgtgtt tctgttccct 900

ccaaagccta aggacaccct gatgatcagc cggacacctg aagtgacctg cgtggtggtg 960

gatgtgtccc acgaggatcc cgaagtgaag ttcaattggt acgtggacgg cgtggaagtg 1020

cacaacgcca agaccaagcc tagagaggaa cagtacaaca gcacctacag agtggtgtct 1080

gtgctgacag tgctgcacca ggactggctg aacggcaaag agtacaagtg caaggtgtcc 1140

aacaaggccc tgcctgctcc tatcgagaaa accatcagca aggccaaggg ccagcctagg 1200

gaaccccagg tttacaccct gcctccatgc agggatgagc tgaccaagaa ccaggtgtcc 1260

ctgtggtgcc tggttaaggg cttctacccc tccgatatcg ccgtggaatg ggagagcaat 1320

ggccagccag agaacaacta caagacaacc cctcctgtgc tggactccga cggctcattc 1380

ttcctgtaca gcaagctgac cgtggacaag agcagatggc agcagggcaa cgtgttcagc 1440

tgcagcgtga tgcacgaggc cctgcacaac cactacacac agaagtccct gtctctgagc 1500

cccggcaaa 1509

<210> 194

<211> 503

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0015-HC-k

<400> 194

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Gly Gly Gly Gly Thr Tyr Tyr

65 70 75 80

Ser Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys

85 90 95

Asp Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Val Phe Gly Val Val Thr Pro Tyr

115 120 125

Tyr Tyr Phe Ala Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val

130 135 140

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

145 150 155 160

Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val

165 170 175

Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Asn Ser

180 185 190

Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu

195 200 205

Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser

210 215 220

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln

225 230 235 240

Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Ala Pro

245 250 255

Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Gly Glu

260 265 270

Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

275 280 285

Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

290 295 300

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

305 310 315 320

Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp

325 330 335

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr

340 345 350

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

355 360 365

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu

370 375 380

Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

385 390 395 400

Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys

405 410 415

Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

420 425 430

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

435 440 445

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

450 455 460

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

465 470 475 480

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

485 490 495

Leu Ser Leu Ser Pro Gly Lys

500

<210> 195

<211> 1512

<212> DNA

<213> Artificial

<220>

<223> Full-length nucleotide sequence of NYC-0016-HC-k

<400> 195

atgaagcacc tgtggttctt tctgctgctg gtggccgctc ctagatgggt gctgtctggc 60

gaagtgcagc tggtggaatc tggtggcgga gttgttcagc ctggcggctc tctgagactg 120

agctgtgctg tgtccggctt caccttcgac gactacgcca tgcactgggt ccgacaggca 180

cctggcaaag gccttgaatg ggtgtccctg atcagcggag atggcggcag cacacactac 240

gccgattctg tgaagggcag attcaccatc agccgggaca acagcaagaa cagcctgtac 300

ctgcagatga actccctgcg gacaggcgac acagccctgt actactgcgc caaggacatg 360

atcttcgccg tggtcatcac cgactaccac tactacggca tggacgtgtg gggccaggga 420

acccacagtga cagtttctag cggaggcgga ggaagtggcg gcggaggatc tggcggtggt 480

ggttctgata tccagatgac acagagcccc agcagcctgt ctgcctctgt ggggacaga 540

gtgaccatca cctgtagagc cagccagagc atcagcagct acctgaactg gtatcagcag 600

aagcccggca aggcccctaa actgctgatc tatgccgcct ccagtctgca gagcggagtg 660

cctagcagat tttctggcag cggctccggc accgatttca ccctgaccat atctagcctg 720

cagcctgagg acttcgccac ctactactgc cagcagagct acagcacccc tcctatcacc 780

tttggccagg gcaccagact ggaaatcaaa ggcggagaac ccaagagcgc agacaagacc 840

cacacctgtc ctccatgtcc tgctccagaa gctgcaggcg gcccttccgt gtttctgttc 900

cctccaaagc ctaaggacac cctgatgatc agccggacac ctgaagtgac ctgcgtggtg 960

gtggatgtgt cccacgagga tcccgaagtg aagttcaatt ggtacgtgga cggcgtggaa 1020

gtgcacaacg ccaagaccaa gcctagagag gaacagtaca acagcaccta cagagtggtg 1080

tctgtgctga cagtgctgca ccaggactgg ctgaacggca aagagtacaa gtgcaaggtg 1140

tccaacaagg ccctgcctgc tcctatcgag aaaaccatca gcaaggccaa gggccagcct 1200

agggaacccc aggtttacac cctgcctcca tgcagggatg agctgaccaa gaaccaggtg 1260

tccctgtggt gcctggttaa gggcttctac ccctccgata tcgccgtgga atggggagagc 1320

aatggccagc cagagaacaa ctacaagaca acccctcctg tgctggactc cgacggctca 1380

ttcttcctgt acagcaagct gaccgtggac aagagcagat ggcagcaggg caacgtgttc 1440

agctgcagcg tgatgcacga ggccctgcac aaccactaca cacagaagtc cctgtctctg 1500

agccccggca aa 1512

<210> 196

<211> 504

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYC-0016-HC-k

<400> 196

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr

35 40 45

Phe Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ser Leu Ile Ser Gly Asp Gly Gly Ser Thr His Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Thr Gly Asp Thr Ala

100 105 110

Leu Tyr Tyr Cys Ala Lys Asp Met Ile Phe Ala Val Val Ile Thr Asp

115 120 125

Tyr His Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr

130 135 140

Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser

165 170 175

Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser

180 185 190

Ser Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu

195 200 205

Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe

210 215 220

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

225 230 235 240

Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr

245 250 255

Pro Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Gly

260 265 270

Glu Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala

275 280 285

Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

290 295 300

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

305 310 315 320

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val

325 330 335

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

340 345 350

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

355 360 365

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala

370 375 380

Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

385 390 395 400

Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr

405 410 415

Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser

420 425 430

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

435 440 445

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

450 455 460

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

465 470 475 480

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

485 490 495

Ser Leu Ser Leu Ser Pro Gly Lys

500

<210> 197

<211> 719

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYZ-1007-HC2

<400> 197

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val

20 25 30

Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Glu Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

385 390 395 400

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

405 410 415

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

420 425 430

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

435 440 445

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

450 455 460

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

465 470 475 480

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

485 490 495

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

500 505 510

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

515 520 525

Pro Glu Val Thr Cys Val Val Val Ser Val Ser His Glu Asp Pro Glu

530 535 540

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

545 550 555 560

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

565 570 575

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

580 585 590

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

595 600 605

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro

610 615 620

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

625 630 635 640

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

645 650 655

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

660 665 670

Asp Gly Ser Phe Ala Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg

675 680 685

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

690 695 700

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

705 710 715

<210> 198

<211> 719

<212> PROTEIN

<213> Artificial

<220>

<223> Full-length amino acid sequence of NYZ-1017-HC2

<400> 198

Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp

1 5 10 15

Val Leu Ser Gly Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val

20 25 30

Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser

35 40 45

Ile Arg Ser Tyr Asp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly

50 55 60

Leu Glu Trp Val Ala Thr Ile Ser Tyr Asp Gly Ser Gln Lys Tyr Tyr

65 70 75 80

Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys

85 90 95

Asn Thr Leu Tyr Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala

100 105 110

Val Tyr His Cys Ala Arg Gly Ser Ser Gly His Tyr Glu Ala Phe Asp

115 120 125

Ile Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Ser Val Leu Thr

145 150 155 160

Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys Val Thr Ile Ser

165 170 175

Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Trp Tyr Val Ser Trp Tyr

180 185 190

Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile Tyr Asp Asn Asn

195 200 205

Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Lys Ser Gly

210 215 220

Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr Gly Asp Glu Ala

225 230 235 240

Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser Ala Pro Trp Val

245 250 255

Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly Gly Gly Gly Ser Glu

260 265 270

Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser

275 280 285

Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Thr Phe Asn Tyr Tyr Gly

290 295 300

Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala

305 310 315 320

Ser Ile Thr Arg Ser Gly Gly Arg Ile Tyr Tyr Pro Asp Ser Val Lys

325 330 335

Gly Arg Phe Thr Ile Ser Arg Glu Asn Thr Gln Lys Thr Leu Tyr Leu

340 345 350

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr

355 360 365

Leu Asp Gly Arg Asp Gly Trp Val Ala Tyr Trp Gly Gln Gly Thr Leu

370 375 380

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

385 390 395 400

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

405 410 415

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

420 425 430

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

435 440 445

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

450 455 460

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

465 470 475 480

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

485 490 495

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

500 505 510

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

515 520 525

Pro Glu Val Thr Cys Val Val Val Ser Val Ser His Glu Asp Pro Glu

530 535 540

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

545 550 555 560

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

565 570 575

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

580 585 590

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

595 600 605

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Val Tyr Pro

610 615 620

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

625 630 635 640

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

645 650 655

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

660 665 670

Asp Gly Ser Phe Ala Leu Val Ser Lys Leu Thr Val Asp Lys Ser Arg

675 680 685

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

690 695 700

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

705 710 715

230

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Claims (132)

1. An antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof, comprising: CDRH1 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:54, CDRH2 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:55, CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:56, a CDRL1 light chain consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:57, wherein amino acid 7 is W, CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:58, and CDRL3 light chain consisting of the amino acid sequence of SEQ ID NO:59. 2. An antibody, or a binding fragment thereof, according to claim 1, comprising a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:53, and a light chain variable region consisting of the amino acid sequence of amino acids 156-266 of the amino acid sequence of SEQ ID NO:53, or a heavy chain variable region consisting of the amino acid sequence of amino acids 21-140 of the amino acid sequence of SEQ ID NO:156, and a light chain variable region consisting of the amino acid sequence of amino acids 161-271 of the amino acid sequence SEQ ID NO:156. 3. An antibody, or a binding fragment thereof, according to claim 1 or 2, which is an scFv; scFv consists of the amino acid sequence of amino acids 21-266 of the amino acid sequence of SEQ ID NO:53, or scFv consists of the amino acid sequence of amino acids 21-271 of the amino acid sequence SEQ ID NO:156. 4. A polynucleotide encoding an antibody or a binding fragment thereof according to any one of claims 1-3. 5. An expression vector containing a polynucleotide according to item 4. 6. A host cell for producing an antibody or a binding fragment thereof according to any one of claims 1 to 3, wherein the cell contains the polynucleotide according to item 4 or the vector according to item 5. 7. A method for producing an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof, according to claim 1, comprising: (i) a step of culturing the host cell according to claim 6; and (ii) a step of purifying the antibody or its binding fragment from the culture product obtained in step (i). 8. An antibody or a binding fragment thereof according to claim 1, which is obtained by the method according to claim 7. 9. A pharmaceutical composition for the prevention or treatment of cancer with cells expressing HLA-A2/NY-ESO, containing as an active ingredient an antibody or a binding fragment thereof, according to any one of claims 1-3 and 8, a polynucleotide according to claim 4, a vector according to claim 5, or a cell according to claim 6. 10. A bispecific antibody that specifically binds to human HLA/NY-ESO and comprises an antibody, or a binding fragment thereof, according to any one of claims 1-3 and 8, and specifically binds to CD3 and comprises an antibody that specifically binds to CD3 or a binding fragment thereof, wherein the antibody that specifically binds to CD3 or a binding fragment thereof comprises: (CCH1) CDRH1 heavy chain consisting of the amino acid sequence of SEQ ID NO:141; (CCH2) CDRH2 of the heavy chain consisting of the amino acid sequence of SEQ ID NO:142, or a heavy chain CDRH2 consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:142, wherein amino acid 3 is N or S; (CCH3) CDRH3 of the heavy chain, consisting of the amino acid sequence of SEQ ID NO:143; (CCL1) CDRL1 light chain consisting of the amino acid sequence of SEQ ID NO:144; (CCL2) CDRL2 light chain consisting of the amino acid sequence of SEQ ID NO:145, or a CDRL2 light chain consisting of an amino acid sequence derived from the amino acid sequence of SEQ ID NO:145, wherein amino acid 2 is N; and (CCL3) CDRL3 light chain consisting of the amino acid sequence of SEQ ID NO:146. 11. The bispecific antibody of claim 10, wherein the antibody that specifically binds to CD3, or a binding fragment thereof, comprises: (CH1CL1) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:136, and a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:136; (CH2CL2) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:137, and a light chain variable region consisting of the amino acid sequence of amino acids 135-241 of the amino acid sequence SEQ ID NO:137; (CH3CL3) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:147, and a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:147; (CH4CL4) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:138, and a light chain variable region consisting of the amino acid sequence of amino acids 135-241 of the amino acid sequence SEQ ID NO:138; (CH5CL5) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:139, and a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:139; (CH6CL6) a heavy chain variable region consisting of the amino acid sequence of amino acids 2-119 of the amino acid sequence of SEQ ID NO:140, and a light chain variable region consisting of the amino acid sequence of amino acids 135-243 of the amino acid sequence SEQ ID NO:140; (CH7CL7) a heavy chain variable region consisting of the amino acid sequence of amino acids 272-389 of the amino acid sequence of SEQ ID NO:155, and a light chain variable region consisting of the amino acid sequence of amino acids 405-511 of the amino acid sequence SEQ ID NO:155; (CH8CL8) a heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:156, and a light chain variable region consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence of SEQ ID NO:156; or (CH9CL9) a heavy chain variable region consisting of the amino acid sequence of amino acids 277-394 of the amino acid sequence of SEQ ID NO:157, and a light chain variable region consisting of the amino acid sequence of amino acids 410-516 of the amino acid sequence SEQ ID NO:157. 12. The bispecific antibody of claim 11, wherein the antibody that specifically binds to CD3, or a binding fragment thereof, is an scFv; preferably, wherein the scFv comprises: (CS1) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:136; (CS2) an scFv consisting of the amino acid sequence of amino acids 2-241 of the amino acid sequence of SEQ ID NO:137; (CS3) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:147; (CS4) an scFv consisting of the amino acid sequence of amino acids 2-241 of the amino acid sequence of SEQ ID NO:138; (CS5) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:139; (CS6) an scFv consisting of the amino acid sequence of amino acids 2-243 of the amino acid sequence of SEQ ID NO:140; (CS7) an scFv consisting of the amino acid sequence of amino acids 272-511 of the amino acid sequence of SEQ ID NO:155; (CS8) an scFv consisting of the amino acid sequence of amino acids 277-516 of the amino acid sequence of SEQ ID NO:156; or (CS9) scFv consisting of the amino acid sequence of amino acids 277-516 of the amino acid sequence SEQ ID NO:157. 13. A bispecific antibody according to claim 11 or 12, comprising a first polypeptide, wherein the first polypeptide comprises an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof, an antibody that specifically binds to CD3, or an antigen-binding fragment thereof; and an Fc region (i) in order from the N terminus to the C terminus; wherein the antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof, is an scFc, wherein the antibody that specifically binds to CD3, or a binding fragment thereof, is an scFc; and a second polypeptide comprising an Fc region (ii), where the first polypeptide is linked to the second polypeptide in Fc region (i) and Fc region (ii). 14. The bispecific antibody of claim 13, wherein the scFv that specifically binds to human HLA/NY-ESO comprises an amino acid sequence consisting of the amino acid sequence of amino acids 21-266 of SEQ ID NO:53, or an amino acid sequence consisting of the amino acid sequence of amino acids 21-271 of SEQ ID NO:156. 15. A bispecific antibody according to claim 13 or 14, which comprises an amino acid sequence selected from the group consisting of amino acid sequence from amino acids 21-511 of the amino acid sequence SEQ ID NO:96, the amino acid sequence of amino acids 21-516 of the amino acid sequence SEQ ID NO:156, and amino acid sequence from amino acids 21-516 of the amino acid sequence SEQ ID NO:157. 16. A bispecific antibody according to any one of claims 13-15, wherein the first polypeptide comprises an amino acid sequence selected from the group consisting of amino acid sequence from amino acids 529-745 of the amino acid sequence SEQ ID NO:96, the amino acid sequence of amino acids 534-750 of the amino acid sequence of SEQ ID NO:156, and an amino acid sequence of amino acids 534-750 of the amino acid sequence of SEQ ID NO:157; or amino acid sequence from amino acids 20-745 of the amino acid sequence SEQ ID NO:96, an amino acid sequence of amino acids 20-750 of the amino acid sequence SEQ ID NO:156, and amino acid sequence from amino acids 20-750 of the amino acid sequence SEQ ID NO:157. 17. A bispecific antibody according to any one of claims 13-16, wherein the second polypeptide comprises an amino acid sequence of amino acids 20-246 of the amino acid sequence SEQ ID NO:84. 18. A bispecific antibody according to claim 11 or 12, comprising first polypeptide, wherein the first polypeptide comprises an antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof, an antibody that specifically binds to CD3, or a binding fragment thereof, and an Fc region (i) in order from the N terminus to the C terminus, wherein the antibody that specifically binds to CD3, or a binding fragment thereof, comprises a heavy chain variable region and a CH1 constant region, the second polypeptide and the third polypeptide, wherein the antibody that specifically binds to human HLA/NY-ESO, or a binding fragment thereof, is an scFv, and an antibody that specifically binds to CD3, or a binding fragment thereof, is an scFv, the second polypeptide comprises an immunoglobulin heavy chain containing an Fc region (ii), and the third polypeptide comprises an immunoglobulin light chain; and the second polypeptide is associated with the third polypeptide, and the first polypeptide is associated with the second polypeptide in the Fc region, the second polypeptide comprises an immunoglobulin hinge region and an immunoglobulin Fc region (ii); and the third polypeptide comprises an antibody light chain consisting of a variable region and a constant region, wherein the first polypeptide is linked to a second polypeptide in Fc region (i) and Fc region (ii), and the first polypeptide is linked to a third polypeptide in the variable region of the heavy chain and the constant region of the CH1 antibody. 19. The bispecific antibody of claim 17, wherein the antibody that specifically binds to CD3 is a Fab. 20. The bispecific antibody of claim 18 or 19, wherein the first polypeptide comprises an amino acid sequence of amino acids 21-394 of the amino acid sequence SEQ ID NO:160, an amino acid sequence of amino acids 20-724 of the amino acid sequence SEQ ID NO:160; or amino acid sequence from amino acids 20-719 of the amino acid sequence SEQ ID NO:197. 21. A bispecific antibody according to any one of claims 18-20, wherein the second polypeptide comprises an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84 and/or wherein the third polypeptide comprises an amino acid sequence of amino acids 21-127 of the amino acid sequence SEQ ID NO:161; preferably, wherein the second polypeptide comprises an amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO: 84, and wherein the third polypeptide comprises an amino acid sequence of amino acids 21-233 of the amino acid sequence of SEQ ID NO: 161, more preferably, wherein the first polypeptide comprises an amino acid sequence of amino acids 20-724 of the amino acid sequence of SEQ ID NO:160; or an amino acid sequence of amino acids 20-719 of the amino acid sequence of SEQ ID NO:197; and wherein the second polypeptide comprises the amino acid sequence of amino acids 20-246 of the amino acid sequence of SEQ ID NO:84, and wherein the third polypeptide comprises an amino acid sequence of amino acids 21-233 of the amino acid sequence SEQ ID NO:161. 22. A bispecific antibody according to any one of claims 10-21, wherein 1 or 2 amino acids are removed from the carboxyl terminus of the amino acid sequence of at least one polypeptide included in the molecule. 23. A nucleic acid molecule encoding a bispecific antibody according to any one of claims 10 to 22. 24. An expression vector containing a nucleic acid molecule according to claim 23. 25. A host cell for producing a bispecific antibody according to any one of claims 10-21, wherein the cell contains the vector according to claim 24. 26. A method for producing a bispecific antibody that specifically binds to human HLA/NY-ESO and human CD3, according to claim 10, comprising: (i) a step of culturing the host cell according to claim 25; and (ii) a step of purifying the antibody, or a binding fragment thereof, from the culture product obtained in step (i). 27. A bispecific antibody that specifically binds to human HLA/NY-ESO and human CD3, according to claim 10, which is obtained by the method according to claim 26. 28. A pharmaceutical composition for the prevention or treatment of cancer with cells expressing HLA-A2/NY-ESO, containing as an active ingredient a bispecific antibody according to any one of claims 10-22 and 27, a polynucleotide according to claim 23, a vector according to claim 24, or a host cell according to claim 25. 29. A pharmaceutical composition according to claim 9 or 28 for use in the prevention or treatment of cancer, preferably, wherein the cancer is one or more cancers selected from the group consisting of renal cancer, melanoma, squamous cell carcinoma, basal cell carcinoma, conjunctival cancer, oral cavity cancer, laryngeal cancer, pharyngeal cancer, thyroid cancer, lung cancer, breast cancer, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, colon cancer, rectal cancer, appendiceal cancer, anal cancer, liver cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, adrenal cancer, bladder cancer, prostate cancer, uterine cancer, vaginal cancer, liposarcoma, angiosarcoma, chondrosarcoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, undifferentiated pleomorphic sarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, retroperitoneal sarcoma, synovial sarcoma, uterine sarcoma, gastrointestinal stromal tumor, leiomyosarcoma, epithelioid sarcoma, B-cell lymphoma, T/NK-cell lymphoma, Hodgkin's lymphoma, myelogenous leukemia, lymphatic leukemia, myeloproliferative disease, myelodysplastic syndrome, multiple myeloma, testicular carcinoma and ovarian cancer; more preferably, wherein the lung cancer is non-small cell lung cancer and small cell lung cancer; even more preferably, wherein the non-small cell lung cancer is adenocarcinoma, epidermoid carcinoma or large cell carcinoma; and, even more preferably, where the composition is used in combination with another agent. 30. A bispecific antibody according to any one of claims 10-22 and 27 for use in the prevention or treatment of cancer, preferably, wherein the cancer is one or more cancers selected from the group consisting of renal cancer, melanoma, squamous cell carcinoma, basal cell carcinoma, conjunctival cancer, oral cavity cancer, laryngeal cancer, pharyngeal cancer, thyroid cancer, lung cancer, breast cancer, esophageal cancer, gastric cancer, duodenal cancer, small intestine cancer, colon cancer, rectal cancer, appendiceal cancer, anal cancer, liver cancer, gallbladder cancer, bile duct cancer, pancreatic cancer, adrenal cancer, bladder cancer, prostate cancer, uterine cancer, vaginal cancer, liposarcoma, angiosarcoma, chondrosarcoma, rhabdomyosarcoma, Ewing's sarcoma, osteosarcoma, undifferentiated pleomorphic sarcoma, myxofibrosarcoma, malignant peripheral nerve sheath tumor, retroperitoneal sarcoma, synovial sarcoma, uterine sarcoma, gastrointestinal stromal tumor, leiomyosarcoma, epithelioid sarcoma, B-cell lymphoma, T/NK-cell lymphoma, Hodgkin's lymphoma, myelogenous leukemia, lymphatic leukemia, myeloproliferative disorder, myelodysplastic syndrome, multiple myeloma, testicular carcinoma, and ovarian cancer. more preferably, wherein the lung cancer is non-small cell lung cancer and small cell lung cancer; even more preferably, wherein the non-small cell lung cancer is adenocarcinoma, epidermoid carcinoma or large cell carcinoma; and, even more preferably, where the bispecific antibody is used in combination with another agent.

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