Anti-PD-L2 antibodies

Abstract

The present invention relates to anti-PD-L2 antibodies and antigen-binding fragments thereof, such as mouse, chimeric and humanized monoclonal antibodies and antigen-binding fragments thereof. The present invention also provides nucleic acids encoding the above antibodies and antigen-binding fragments thereof, as well as expression vectors and cells for expressing the antibodies and antigen-binding fragments thereof. Methods of using such antibodies and antigen-binding fragments thereof are also provided, for example, the present invention provides methods of using chimeric and humanized anti-PD-L2 antibodies and antigen-binding fragments thereof.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and in particular relates to antibodies and antigen-binding fragments thereof that specifically bind to PD-L2, and to methods for using such antibodies and antigen-binding fragments. Background Art

[0002] Tumor immunotherapy is an important anti-tumor treatment method, and its core is to activate the anti-tumor response of T lymphocytes in tumor patients and enhance their killing function against tumor cells. Programmed death receptor 1 (PD-1) is a protein receptor expressed on immune cells (including T cells and B cells). It has two ligands, namely programmed death-1 ligand 1 (PD-L1, B7-H1 or CD274) and programmed death-1 ligand 2 (PD-L2, B7-DC or CD273). After binding to the ligand, PD-1 can transmit inhibitory signals, negatively regulate T cell immune responses, and inhibit TCR-mediated T cell activation, cytokine production, and T cell proliferation. Therefore, the role of PD-1 / PD-L1 and PD-1 / PD-L2 signaling pathways in tumor immune escape and their application in tumor immunotherapy have become research hotspots.

[0003] Both PD-L2 and PD-L1 are type I transmembrane proteins composed of an IgV-like domain, an IgC-like domain, a transmembrane region, and a tail cytoplasmic region. The genes encoding human PD-L2 and PD-L1 are both located on chromosome 9p24.2, but the sequences, tissue distribution, receptor-ligand binding specificity and affinity, and signaling pathways involved in PD-L2 and PD-L1 show great differences. PD-L1 is widely distributed on leukocytes and non-hematopoietic cells in lymphoid and non-lymphoid tissues and various cancer cells, and PD-L1 is upregulated during tumorigenesis; PD-L2 is generally expressed on antigen-presenting cells, including dendritic cells, macrophages, and bone marrow-derived mast cells (Zhong et al. (2007) Eur. J. Immunol 37: 2405-2410). Under the stimulation of a specific microenvironment, PD-L2 can be induced to express in a variety of immune cells and non-immune cells, such as head and neck squamous cell carcinoma, breast cancer, gastric cancer and other tumor cells (Zou et al. (2015) Nat Rev Immunol 8:467-477). This shows that blocking the PD-1 / PD-L2 interaction can also enhance tumor-specific T cell immunity and help the immune system to eliminate tumor cells. More importantly, high expression of PD-L2 was detected in a variety of human tumor samples that do not express or low-express PD-L1 (Baptista et al. (2016) Humanpathology 47:78-84; Yearley et al. (2017) Clin cancer research 23:3158-3167). In addition, among 172 patients with recurrent or metastatic squamous cell carcinoma of the neck who received Keytruda treatment, the ORR of patients with positive expression of both PD-L1 and PD-L2 was 27.5%, while the ORR of patients with positive PD-L1 and negative PD-L2 was only 11.4%, which is twice that of the latter (Yearley et al. (2017) Clin cancer research 23:3158-3167), indicating that simultaneous inhibition of PD-L1 and PD-L2 may improve the clinical response rate, and PD-L2 can serve as a supplementary indicator for predicting the clinical efficacy of PD-1 inhibitors.

[0004] Since 2010, a variety of antibodies targeting PD-1 or PD-L1 have entered clinical trials or obtained marketing approval. These antibodies have shown significant anti-tumor activity both in vitro and in vivo, such as Pembrolizumab, Atezolizumab, and Durvalumab, but so far no antibody drugs targeting PD-L2 have entered clinical research. Therefore, it is of great significance to develop a therapeutic antibody that specifically binds to human PD-L2. In addition, PD-L2, as a molecular marker, plays an important role in the targeted therapy and prognostic diagnosis of various tumors. Therefore, it is necessary to prepare a monoclonal antibody that highly specifically binds to human PD-L2 to guide the diagnosis and treatment of tumors. Summary of the invention

[0005] On the one hand, the present invention provides an isolated antibody or antigen-binding fragment thereof, which specifically binds to programmed death-1 ligand 2 (PD-L2). In some embodiments, the isolated antibody or antigen-binding fragment thereof of the present invention is a murine antibody, a chimeric antibody, a humanized antibody or a human antibody. In some embodiments, the isolated antibody or antigen-binding fragment thereof of the present invention is a monoclonal antibody, a Fab fragment, a F(ab')2 fragment, a Fd fragment, a Fv fragment, a dAb, an isolated CDR, a single-chain Fv molecule, and a recombinant polypeptide, a fusion protein, a bispecific molecule or a combination thereof. In some embodiments, the isolated antibody or antigen-binding fragment thereof of the present invention binds to human PD-L2; in other embodiments, the isolated antibody or antigen-binding fragment thereof of the present invention binds to human PD-L2 and binds to cynomolgous PD-L2. In another embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention blocks the binding of PD-L2 to its receptor PD-1.

[0006] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3, wherein (1) the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 1, 2, 3, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs: 1, 2, 3; (2) the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 7, 8, 9, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs: 7, 8, 9, respectively. NOs:7, 8, 9 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences set forth in SEQ ID NOs:7, 8, 9; (3) the heavy chain CDR1, heavy chain CDR2 and heavy chain CDR3 respectively comprise the amino acid sequences set forth in SEQ ID NOs:13, 14, 15 or are identical to the amino acid sequences set forth in SEQ ID NOs:14, 16, 17 or are identical to the amino acid sequences set forth in SEQ ID NOs:15, 18, 19 or are identical to the amino acid sequences set forth in SEQ ID NOs:16, 19, 20 or are identical to the amino acid sequences set forth in SEQ ID NOs:17, 18, 21 or are identical to the amino acid sequences set forth in SEQ ID NOs:18, 19, 22 or are identical to the amino acid sequences set forth in SEQ ID NOs:19, 23, 24 or are identical to the amino acid sequences set forth in SEQ ID NOs:19, 25, 26 or are identical to the amino acid sequences set forth in SEQ ID NOs:19, or (4) the heavy chain CDR1, heavy chain CDR2 and heavy chain CDR3 comprise the amino acid sequences of SEQ ID NOs:19,20,21, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences of SEQ ID NOs:19,20,21, respectively;

[0007] Among them, the amino acid sequence of SEQ ID NO: 2 is RIDPSXSETH, X=D or E; the amino acid sequence of SEQ ID NO: 8 is MIDPSXSETR, X=D or E; the amino acid sequence of SEQ ID NO: 14 is VINPVXGGIS, X=N or Q; the amino acid sequence of SEQ ID NO: 20 is LISPYXGVTT, X=N or Q.

[0008] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein (1) the light chain CDR1, the light chain CDR2 and the light chain CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 4, 5, 6, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs: 4, 5, 6; (2) the light chain CDR1, the light chain CDR2 and the light chain CDR3 comprise the amino acid sequences set forth in SEQ ID NOs: 10, 11, 12, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs: 10, 11, 12, respectively The amino acid sequences of SEQ ID NOs: 10, 11, 12 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity; (3) the light chain CDR1, light chain CDR2 and light chain CDR3 respectively comprise the amino acid sequences of SEQ ID NOs: 16, 17, 18 or are identical to the amino acid sequences of SEQ ID NOs: 19, 20, 21, 22 or are identical to the amino acid sequences of SEQ ID NOs: 23, 24, 25 or are identical to the amino acid sequences of SEQ ID NOs: 31, 32, 33, 34, 35 or (4) the light chain CDR1, light chain CDR2 and light chain CDR3 comprise the amino acid sequences of SEQ ID NOs:22, 23, 24, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences of SEQ ID NOs:22, 23, 24, respectively;

[0009] Among them, the amino acid sequence of SEQ ID NO: 22 is KSSQSLLXSGNQKNYLT, X=N or Q.

[0010] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3, and a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the heavy chain CDR1 comprises an amino acid sequence as set forth in SEQ ID NOs: 1, 7, 13, or 19, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence as set forth in SEQ ID NOs: 1, 7, 13, or 19; the heavy chain CDR2 comprises an amino acid sequence as set forth in SEQ ID NOs: 2, 8, 14, or 20, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence as set forth in SEQ ID NOs: 2, 8, 14, or 20 NOs: 2, 8, 14, or 20 has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence shown in SEQ ID NOs: 3, 9, 15, or 21; the heavy chain CDR3 comprises the amino acid sequence shown in SEQ ID NOs: 3, 9, 15, or 21 or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence shown in SEQ ID NOs: 3, 9, 15, or 21; and the light chain CDR1 comprises SEQ ID NOs: NOs:4, 10, 16, or 22, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NOs:4, 10, 16, or 22; the light chain CDR2 comprises the amino acid sequence of SEQ ID NOs:5, 11, 17, or 23, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NOs:4, 10, 16, or 22; An amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequence set forth in NOs: 5, 11, 17, or 23;the light chain CDR3 comprises an amino acid sequence as set forth in SEQ ID NOs: 6, 12, 18, or 24, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence as set forth in SEQ ID NOs: 6, 12, 18, or 24;

[0011] Among them, the amino acid sequence of SEQ ID NO: 2 is RIDPSXSETH, X=D or E; the amino acid sequence of SEQ ID NO: 8 is MIDPSXSETR, X=D or E; the amino acid sequence of SEQ ID NO: 14 is VINPVXGGIS, X=N or Q; the amino acid sequence of SEQ ID NO: 20 is LISPYXGVTT, X=N or Q; the amino acid sequence of SEQ ID NO: 22 is KSSQSLLXSGNQKNYLT, X=N or Q.

[0012] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3, and a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, 3, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 1, 2, 3; and the light chain CDR1, the light chain CDR2 and the light chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 4, 5, 6, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequences shown in SEQ ID NOs: 4, 5, 6, respectively. The amino acid sequences shown in IDNOs: 4, 5, 6 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity;

[0013] Wherein, the amino acid sequence of SEQ ID NO: 2 is RIDPSXSETH, X=D or E.

[0014] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3, and a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 7, 8, 9, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences shown in SEQ ID NOs: 7, 8, 9; and the light chain CDR1, the light chain CDR2 and the light chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 10, 11, 12, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences shown in SEQ ID NOs: 10, 11, 12, respectively. The amino acid sequences shown in IDNOs: 10, 11, 12 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity;

[0015] Wherein, the amino acid sequence of SEQ ID NO: 8 is MIDPSXSETR, X=D or E.

[0016] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3, and a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 13, 14, 15, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences shown in SEQ ID NOs: 13, 14, 15; and the light chain CDR1, the light chain CDR2 and the light chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 16, 17, 18, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences shown in SEQ ID NOs: 16, 17, 18, respectively. The amino acid sequences shown in NOs: 16, 17, 18 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity;

[0017] Among them, the amino acid sequence of SEQ ID NO: 14 is VINPVXGGIS, X=N or Q.

[0018] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain CDR1, a heavy chain CDR2 and a heavy chain CDR3, and a light chain CDR1, a light chain CDR2 and a light chain CDR3, wherein the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 19, 20, 21, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences shown in SEQ ID NOs: 19, 20, 21; and the light chain CDR1, the light chain CDR2 and the light chain CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 22, 23, 24, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences shown in SEQ ID NOs: 22, 23, 24, respectively. The amino acid sequences shown in NOs: 22, 23, 24 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity;

[0019] Among them, the amino acid sequence of SEQ ID NO: 20 is LISPYXGVTT, X=N or Q; the amino acid sequence of SEQ ID NO: 22 is KSSQSLLXSGNQKNYLT, X=N or Q.

[0020] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the invention binds to PD-L2 and comprises a heavy chain variable region comprising an amino acid sequence as set forth in SEQ ID NOs: 25, 27, 29, 31, 33, 35, 36, 38, 39, 41, 42, or 44, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence as set forth in SEQ ID NOs: 25, 27, 29, 31, 33, 35, 36, 38, 39, 41, 42, or 44.

[0021] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the invention binds to PD-L2 and comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NOs: 26, 28, 30, 32, 34, 37, 40, 43, or 45, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence as set forth in SEQ ID NOs: 26, 28, 30, 32, 34, 37, 40, 43, or 45.

[0022] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the invention binds to PD-L2 and comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 25, 27, 29, 31, 33, 35, 36, 38, 39, 41, 42, or 44, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to an amino acid sequence as shown in SEQ ID NOs: 25, 27, 29, 31, 33, 35, 36, 38, 39, 41, 42, or 44; and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NOs: 25, 27, 29, 31, 33, 35, 36, 38, 39, 41, 42, or 44. NOs:26, 28, 30, 32, 34, 37, 40, 43, or 45, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NOs:26, 28, 30, 32, 34, 37, 40, 43, or 45.

[0023] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain variable region and a light chain variable region, wherein (1) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs: 25 and 26, respectively, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 25 and 26; (2) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs: 27 and 28, respectively, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: NO:27 and 28 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NO:29 and 30, respectively; (3) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NO:29 and 30, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:29 and 30, respectively; (4) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NO:29 and 30, respectively. : (5) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs:33 and 34, respectively, or amino acid sequences at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs:33 and 34, respectively;(6) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs: 35 and 34, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 35 and 34; (7) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs: 36 and 37, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 36 and 37, respectively. NO:36 and 37 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NO:38 and 37, respectively; (8) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NO:38 and 37, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:38 and 37, respectively; (9) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NO:39 and 40, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:39 and 40, respectively. NO:39 and 40, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:39 and 40; (10) the heavy chain variable region and the light chain variable region comprise the amino acid sequence of SEQ ID NO:41 and 40, respectively, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:39 and 40, respectively; : (11) the heavy chain variable region and the light chain variable region comprise the amino acid sequences of SEQ ID NOs:42 and 43, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences of SEQ ID NOs:42 and 43, respectively;(12) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs: 44 and 43, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 44 and 43; (13) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NOs: 42 and 45, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 44 and 43, respectively. NO:42 and 45 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NO:42 and 45; or (14) the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in SEQ ID NO:44 and 45, respectively, or amino acid sequences having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NO:44 and 45, respectively.;

[0024] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the invention binds to PD-L2 and comprises a heavy chain comprising an amino acid sequence as set forth in SEQ ID NOs: 46, 50, 54, 58, 62, 64, 68, 70, 74, 76, 80, or 82, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence as set forth in SEQ ID NOs: 46, 50, 54, 58, 62, 64, 68, 70, 74, 76, 80, or 82.

[0025] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the invention binds to PD-L2 and comprises a light chain comprising an amino acid sequence as set forth in SEQ ID NOs: 47, 51, 55, 59, 63, 69, 75, 81, or 83, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence as set forth in SEQ ID NOs: 47, 51, 55, 59, 63, 69, 75, 81, or 83.

[0026] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the invention binds to PD-L2 and comprises a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence as set forth in SEQ ID NOs: 46, 50, 54, 58, 62, 64, 68, 70, 74, 76, 80, or 82, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to an amino acid sequence as set forth in SEQ ID NOs: 46, 50, 54, 58, 62, 64, 68, 70, 74, 76, 80, or 82; and the light chain comprises SEQ ID NOs: 47, 51, 55, 59, 63, 69, 75, 81, or 83, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequence shown in SEQ ID NOs: 47, 51, 55, 59, 63, 69, 75, 81, or 83.

[0027] In one embodiment, the isolated antibody or antigen-binding fragment thereof of the present invention binds to PD-L2 and comprises a heavy chain and a light chain, wherein (1) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs: 46 and 47, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs: 46 and 47; (2) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs: 50 and 51, respectively, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs: 50 and 51 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NOs:50 and 51; (3) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs:54 and 55, respectively, or amino acid sequences at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs:54 and 55; (4) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs:58 and 59, respectively, or amino acid sequences at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs:58 and 59, respectively. NO:58 and 59 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NO:58 and 59; (5) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NO:62 and 63, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:62 and 63; (6) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NO:64 and 63, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:64 The amino acid sequences set forth in NOs: 64 and 63 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity;(7) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs: 68 and 69, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 68 and 69; (8) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs: 70 and 69, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: NO:70 and 69 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequences set forth in SEQ ID NO:70 and 69; (9) the heavy chain and light chain comprise the amino acid sequences set forth in SEQ ID NO:74 and 75, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:74 and 75; (10) the heavy chain and light chain comprise the amino acid sequences set forth in SEQ ID NO:76 and 75, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NO:76 80 and 81, respectively, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 76 and 75; (11) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs: 80 and 81, respectively, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: 80 and 81; (12) the heavy chain and the light chain comprise the amino acid sequences set forth in SEQ ID NOs: 82 and 81, respectively, or an amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences set forth in SEQ ID NOs: The amino acid sequences set forth in NOs: 82 and 81 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity;(13) the heavy chain and the light chain comprise the amino acid sequences of SEQ ID NOs: 80 and 83, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences of SEQ ID NOs: 80 and 83; or (14) the heavy chain and the light chain comprise the amino acid sequences of SEQ ID NOs: 82 and 83, respectively, or amino acid sequences that are at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequences of SEQ ID NOs: The amino acid sequences shown in NO:82 and 83 have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity.

[0028] In some embodiments, the antibodies of the present invention further comprise a heavy chain and a light chain, wherein the heavy chain comprises a heavy chain variable region and a heavy chain constant region, and the light chain comprises a light chain variable region and a light chain constant region, wherein the heavy chain variable region and the light chain variable region respectively comprise the amino acid sequences of the above-mentioned heavy chain variable region and light chain variable region, and the heavy chain constant region can be a human IgG1, IgG2 or IgG4 constant region, preferably an IgG1 constant region; and the light chain constant region can be a human κ constant region or a human λ constant region.

[0029] The antibody of the present invention comprises two heavy chains (H) and two light chains (L) in some embodiments, or is composed of two heavy chains and two light chains, and the heavy chains and light chains are mutually connected by disulfide bonds, wherein each heavy chain comprises the above-mentioned heavy chain variable region (HV), and a heavy chain constant region, wherein the heavy chain variable region (HV) comprises a framework region (FR) and the above-mentioned heavy chain complementary determining region (CDR), each light chain comprises the above-mentioned light chain variable region (LV), and a light chain constant region, wherein the light chain variable region (LV) comprises FR and the above-mentioned light chain CDR, wherein the C-terminus of the heavy chain variable region is connected to the N-terminus of the heavy chain constant region, and the C-terminus of the light chain variable region is connected to the N-terminus of the light chain constant region. The antibody of the present invention can be, for example, a full-length antibody of the IgG1, IgG2 or IgG4 isotype. In another embodiment, the antibody of the present invention can be a single-chain antibody (scFv), or an antibody fragment, such as a Fab, F(ab')2 fragment, a Fd fragment, a Fv fragment, a dAb or an isolated CDR.

[0030] The antibodies or antigen-binding fragments thereof of the present invention specifically bind to PD-L2 (e.g., human PD-L2 and monkey PD-L2) and block the binding of PD-L2 to its receptor PD-1. In some embodiments, the anti-PD-L2 antibodies or antigen-binding fragments thereof provided herein are combined with anti-PD-L1 antibodies, which can block the interaction between PD-L2 / PD-1 and PD-L1 / PD-1, and promote T cell activation, proliferation and / or cytokine production, including but not limited to: IL-2, IFNγ, TNF-α, IL-1, IL-4, IL-5, IL-6, IL-12, IL-17 and GM-CSF. One or more. Therefore, on the other hand, the present invention provides a method for regulating an immune response, comprising contacting T cells and antigen-presenting cells with anti-PD-L2 antibodies or antigen-binding fragments thereof and anti-PD-L1 antibodies provided herein. In one embodiment, the regulation of an immune response by an anti-PD-L2 antibody or antigen-binding fragment thereof provided herein in combination with an anti-PD-L1 antibody can be measured in a mixed lymphocyte (MLR) reaction. In one embodiment, the anti-PD-L2 antibodies provided herein are combined with anti-PD-L1 antibodies to increase the level of cytokine production by lymphocytes in MLR. In one embodiment, the anti-PD-L2 antibodies provided herein are combined with anti-PD-L1 antibodies to increase the level of IL-2 and / or IFN-γ production in MLR.

[0031] In one aspect, the present invention provides an isolated antibody or antigen-binding fragment thereof that binds to PD-L2, wherein the antibody or antigen-binding fragment thereof is produced by a hybridoma selected from the group consisting of hybridomas designated herein as 25G4, 14E7, 24A3, and 7A7. Thus, the present invention also includes antibodies or antigen-binding fragments thereof produced by hybridomas 25G4, 14E7, 24A3, and 7A7, as well as any hybridoma that produces the antibodies disclosed herein.

[0032] In one aspect, the present invention provides antibodies or antigen-binding fragments thereof that bind to the same epitope on PD-L2 as any of the exemplary PD-L2 antibodies or antigen-binding fragments thereof of the present invention. In one embodiment, the present invention provides antibodies or antigen-binding fragments thereof that compete with any of the exemplary PD-L2 antibodies or antigen-binding fragments thereof of the present invention for binding to PD-L2.

[0033] On the other hand, the present invention also provides recombinant polypeptides and fusion proteins, which include one or more antibodies or antigen-binding fragments thereof of the present invention and at least one other functional fragment, wherein the other functional fragments include but are not limited to another peptide, protein or cytokine. On the other hand, the present invention also provides a bispecific molecule, which includes one or more antibodies or antigen-binding fragments thereof of the present invention and at least one other antibody or antigen-binding fragment thereof, and the bispecific molecule can bind to at least two different binding sites or targets. "Bispecific molecules" as used herein encompass molecules with two (i.e., bispecific molecules), three (i.e., trispecific molecules), four (i.e., tetraspecific molecules) or more specificities.

[0034] In one aspect, the present invention also provides a pharmaceutical composition comprising an anti-PD-L2 antibody or an antigen-binding fragment thereof of the present invention, and one or more pharmaceutically acceptable carriers. Optionally, the pharmaceutical composition may further include one or more other antibodies, such as a PD-L1 antibody. In another embodiment, the present invention also provides a pharmaceutical composition comprising a recombinant polypeptide, fusion protein or bispecific molecule of the present invention, and a pharmaceutically acceptable carrier.

[0035] In one aspect, the present invention also provides an isolated nucleic acid encoding an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention. The present invention also provides an expression vector comprising the nucleic acid, and a host cell comprising the expression vector.

[0036] In one aspect, the present invention also provides a method for preparing an anti-PD-L2 antibody or an antigen-binding fragment thereof, comprising the steps of: (i) expressing the antibody or the antigen-binding fragment thereof in a host cell, and (ii) isolating the antibody or the antigen-binding fragment thereof from the host cell or its cell culture.

[0037] In other aspects, the present invention provides a method for regulating an immune response in a subject, the method comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof. In another embodiment, the present invention provides a method for treating, alleviating or preventing cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof.

[0038] In one embodiment, the present invention provides a method for enhancing an anti-tumor immune response in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention, or a pharmaceutical composition thereof. In one embodiment, the present invention provides a method for reducing tumor size or inhibiting tumor cell growth in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention, or a pharmaceutical composition thereof. In one embodiment, the present invention provides a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention, or a pharmaceutical composition thereof. In other embodiments, the method comprises administering to the subject a therapeutically effective amount of a nucleic acid encoding an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention. In other embodiments, the method comprises administering to the subject a therapeutically effective amount of a recombinant polypeptide, fusion protein, or bispecific molecule of the present invention.

[0039] In some embodiments, at least one additional antibody may be administered together with the anti-PD-L2 antibody of the present invention or its antigen-binding portion, such as PD-L1 antibody, PD-1 antibody and / or CTLA-4 antibody, preferably PD-L1 antibody. In some embodiments, at least one additional treatment may be administered together with the anti-PD-L2 antibody of the present invention or its antigen-binding fragment, and the additional treatment includes other tumor treatments such as radiotherapy, chemotherapy, biological products, chemicals, vaccines, etc. In certain embodiments, the subject is a human.

[0040] The tumor or cancer includes, but is not limited to, colorectal cancer, breast cancer, ovarian cancer, pancreatic cancer, gastric cancer, renal cancer, prostate cancer, cervical cancer, myeloma, lymphoma, leukemia, thyroid cancer, endometrial cancer, uterine cancer, bladder cancer, neuroendocrine malignancies, head and neck cancer, salivary gland cancer, thymic cancer, liver cancer, nasopharyngeal cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer, melanoma, basal cell skin cancer, squamous cell skin cancer, dermatofibrosarcoma protuberans, Merkel cell carcinoma, glioblastoma, glioma, sarcoma, mesothelioma and myelodysplastic syndrome, or a combination thereof; and optionally, the tumor is in situ, metastatic, recurrent, and / or refractory.

[0041] In another embodiment, the present invention provides a method for treating an infectious disease in a subject, the method comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof. In other embodiments, the method comprises administering to the subject a therapeutically effective amount of a nucleic acid encoding an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention. The infectious diseases include, but are not limited to, candidiasis, candidemia, aspergillosis, streptococcal pneumonia, streptococcal skin and oropharyngeal disease, gram-negative sepsis, tuberculosis, mononucleosis, influenza, respiratory diseases caused by respiratory syncytial virus, malaria, schistosomiasis, and trypanosomiasis.

[0042] On the other hand, the present invention also provides a method for detecting PD-L2 expression in a sample, comprising contacting the sample with the anti-PD-L2 antibody or antigen-binding fragment of the present invention under conditions allowing formation of a complex between the anti-PD-L2 antibody or antigen-binding fragment of the present invention and PD-L2, and detecting the formation of the complex.

[0043] Based on the following specific description and examples, other features and advantages of the current disclosure will become clearer, and specific description and examples should not be interpreted as limiting. The contents of all documents, Genbank records, patents and published patent applications cited in this application are expressly included in this article by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 Flow cytometry analysis of different concentrations of anti-PD-L2 humanized antibody (Hz25G4-1.1) binding to DC / T cells

[0045] Figure 2a The production of IFN-γ (pg / ml) in response to different concentrations of antibodies in MLR is shown. Figure 2b The production of IL-2 (pg / ml) in response to different concentrations of antibodies in the MLR is shown. Figure 2a and Figure 2b , the antibodies tested are from left to right: anti-PD-L2 antibody (Hz25G4-1.1) + anti-PD-L1 antibody, anti-PD-L1 antibody + anti-PD-L2 antibody (Hz25G4-1.1), anti-PD-L2 antibody (Hz25G4-1.1), anti-PD-L1 antibody and blank control (IgG1), among which the anti-PD-L1 antibody refers to the humanized 5G11-IgG1 in CN107001463, and the amino acid sequences of the heavy chain and light chain are shown in SEQ ID NO: 88 and SEQ ID NO: 89 of the present application. DETAILED DESCRIPTION OF THE INVENTION

[0046] It should be understood that the terms used herein are only for describing specific embodiments and are not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention belongs.

[0047] The term "PD-L2" includes PD-L2 variants, homologs, orthologs and paralogs. For example, in some embodiments, an antibody specific for a human PD-L2 protein may cross-react with a PD-L2 protein of another species (e.g., monkey) in some cases. In other embodiments, an antibody specific for a human PD-L2 protein may be completely specific for a human PD-L2 protein without cross-reacting with other species or other types of proteins, or may cross-react only with PD-L2 proteins of certain other species but not all other species.

[0048] The terms "human PD-L2", "hPD-L2" or "human PD-L2" are used interchangeably herein and refer to a protein having the amino acid sequence of human PD-L2, such as the amino acid sequence of Genbank Accession No. AAI13679. The terms "monkey PD-L2" or "cyno PD-L2" are used interchangeably herein and refer to a protein having the amino acid sequence of monkey PD-L2, such as the amino acid sequence of Uniprot Accession No. G7PSE6.

[0049] The term "antibody" as used herein refers to a binding protein having at least one antigen binding domain. The antibody or antigen binding fragment thereof of the present invention may be a whole antibody or any fragment thereof, including a monoclonal antibody or fragment thereof and an antibody variant or fragment thereof. Examples of antibody fragments include Fab fragments, F(ab')2 fragments, Fv fragments, isolated CDR regions, single-chain Fv (scFv), and any antibody fragments known in the art. Antibodies or fragments thereof also include recombinant polypeptides, fusion proteins, and bispecific molecules. The anti-PD-L2 antibodies or antigen binding fragments thereof disclosed herein may be IgG1, IgG2, IgG3, or IgG4 isotypes. The term "isotype" refers to the type of antibody encoded by the heavy chain constant region gene. In some embodiments, the anti-PD-L2 antibodies or antigen binding fragments thereof disclosed herein are IgG1 isotypes. The anti-PD-L2 antibodies or antigen binding fragments thereof disclosed in the present invention may be derived from any species, including but not limited to mice, rats, rabbits, primates, llamas, and humans. The anti-PD-L2 antibodies or antigen binding fragments thereof of the present invention may be murine antibodies, chimeric antibodies, humanized antibodies, or complete human antibodies.

[0050] An "antigen-binding fragment" or "antibody binding portion" of an antibody refers to one or more fragments of an antibody that retain the function of specifically binding to an antigen (e.g., PD-L2 protein). It has been demonstrated that the antigen-binding function of an antibody can be performed by a fragment of a full-length antibody. Examples encompassed by the term "antigen-binding portion / fragment" of an antibody include: (i) Fab fragment: a fragment consisting of V L 、V H The invention relates to a monovalent fragment consisting of a Fab fragment, a CL fragment and a CH1 domain; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; (v) a dAb fragment consisting of the VH domain (see Ward et al., Nature. 341: 544-546 (1989)); (vi) an isolated complementarity determining region (CDR); and (vii) a nanobody, a heavy chain variable region comprising a single variable domain and two constant domains. In addition, although the two domains VL and VH of the Fv fragment are encoded by different genes, VH and VL can be connected into a single protein chain by a recombinant method through a linker, wherein VL and VH are paired to form a monovalent molecule called single-chain Fv (scFv); see Bird et al., Science.242:423-426 (1988); Huston et al., Proc.Natl.Acad.Sci.85:5879-5883 (1988), these single-chain antibodies are also included in the term antigen binding portion / fragment. These antibody fragments can be obtained by conventional techniques known to those skilled in the art, and the fragments can be functionally screened by the same method as full-length antibodies.

[0051] "Mouse antibody" or "murine antibody" refers to antibodies in which the framework region (FR) and CDR region in the variable region are both derived from mouse germline immunoglobulin sequences. In addition, if the antibody comprises a constant region, the constant region is also derived from mouse germline immunoglobulin sequences. The mouse / murine antibodies of the present invention may include amino acid residues that are not encoded by mouse germline immunoglobulin sequences (e.g., mutations introduced by in vitro random mutations, point mutations, or somatic mutations), but do not include antibodies in which CDR sequences from other mammalian germlines are inserted into mouse framework sequences.

[0052] "Chimeric antibody" refers to an antibody having at least a portion of a heavy chain variable region and at least a portion of a light chain variable region derived from one species; and at least a portion of a constant region derived from another species. For example, in some embodiments, a chimeric antibody may comprise a murine variable region and a human constant region.

[0053] "Humanized antibody" refers to an antibody containing CDRs derived from non-human antibodies and framework regions and constant regions derived from human antibodies. For example, the anti-PD-L2 antibodies provided herein may include CDRs derived from one or more murine antibodies and human framework regions and constant regions. Therefore, in some embodiments, the humanized antibodies provided herein bind to the same epitope on PD-L2 as the murine antibodies from which the CDRs of the antibodies are derived. Exemplary humanized antibodies are provided herein. The humanized antibodies provided by the present invention include anti-PD-L2 antibodies or variants thereof comprising heavy chain CDRs and light chain CDRs provided herein and any other human framework region sequences. The humanized antibodies provided by the present invention include anti-PD-L2 antibodies or variants thereof comprising heavy chain CDRs and light chain CDRs provided herein and framework regions that are structurally similar to the framework region sequences provided herein. Additional modifications may also be made in the framework region to improve the properties of the antibodies provided by the present invention, including chemical modifications, point mutations, or mutations back to residues in the original germline sequence. For example, in some embodiments, one or more amino acids in the human framework regions of the VH and / or VL of the humanized antibodies provided herein are backmutated to the corresponding amino acids in the parent murine antibody.

[0054] As used herein, "derived", when used in reference to a molecule or polypeptide relative to a reference antibody or other binding protein, means a molecule or polypeptide that is capable of specifically binding to the same epitope as the reference antibody or other binding protein.

[0055] "Isolated" refers to a compound of interest (eg, an antibody, antigen-binding fragment, or nucleic acid) that has been separated from its natural environment.

[0056] "Antibodies that specifically bind to an antigen" and "antibodies specific for an antigen" are used interchangeably herein with the term "antibodies that specifically bind to an antigen". An antibody that "specifically binds to human PD-L2" refers to an antibody that binds to human PD-L2 protein (and possibly PD-L2 protein of other non-human species) but does not substantially bind to non-PD-L2 proteins. Preferably, the antibody binds to human PD-L2 with "high affinity", i.e., a KD of 5.0×10 -8 M or smaller, 1.0×10 -8 M or less, preferably 5.0×10 -9 M or less, more preferably 1.0×10 -9 M or smaller.

[0057] The term "substantially not binding" to a protein or cell means not binding to the protein or cell, or not binding to the protein or cell with high affinity, i.e., the KD for binding to the protein or cell is 1.0×10 -6 M or more, preferably 1.0×10 -5 M or higher, 1.0×10 -4M or more, more preferably 1.0×10 -3 M or higher, 1.0×10 -2 M or higher.

[0058] The term "high affinity" for IgG means that the KD for binding to antigen is 1.0×10 -6 M or smaller, 1.0×10 -7 M or less, preferably 1.0×10 -8 M or smaller, 5.0×10 -9 M or less, more preferably 1.0×10 -9 M or less. However, for other antibody isotypes, “high affinity” binding may be different. For example, “high affinity” binding for an IgM isotype is defined as binding to an antigen with a KD of 1.0 × 10 -6 M or less, preferably 1.0×10 -7 M or less, more preferably 1.0×10 -8 M or smaller.

[0059] "Identity" refers to the similarity between two nucleic acid sequences or between two polypeptide sequences. The sequence identity of the present invention is at least 85%, 90% or 95%, preferably at least 95%. Non-limiting examples include: 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%. The sequence comparison and identity percentage determination between two sequences can be performed by the default settings of the BLASTN / BLASTP algorithm on the National Center For Biotechnology Institute website.

[0060] "Competitive binding" antibodies refer to antibodies that partially or completely block other antibodies from binding to the target. Whether two antibodies compete with each other for binding to the target, i.e., whether one antibody blocks the binding of another antibody to the target and to what extent, can be determined using competition experiments known in the art, such as solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assays, etc. In certain embodiments, an antibody competes with another antibody for binding to the target and blocks the binding of the other antibody to the target by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.

[0061] Two or more antibodies "bind to the same epitope" means that the antibodies bind to the same stretch of amino acid residues as determined by a given method. Techniques for determining whether an antibody binds to "the same epitope on PD-L2" as an antibody described herein include, for example, epitope mapping methods, such as x-ray analysis of crystals of the antigen: antibody complex and hydrogen / deuterium exchange mass spectrometry (HDX-MS).

[0062] The term "EC50", also known as the half effective concentration, refers to the antibody concentration that achieves 50% of the maximum effect after a specific exposure time. The term "IC50", also known as the half inhibitory concentration, refers to the antibody concentration that inhibits a specific biological or biochemical function by 50% relative to the absence of the antibody. Both EC50 and IC50 can be measured by ELISA or FACS analysis or any other method known in the art.

[0063] "KD" refers to the equilibrium dissociation constant, which is obtained from the ratio of kd to ka (i.e., kd / ka) and is expressed as a molar concentration (M). The KD value of an antibody can be determined using methods well established in the art. A preferred method for determining the KD of an antibody is by using surface plasmon resonance technology, preferably using a biosensor system such as Surface plasmon resonance system analysis.

[0064] "Patient" or "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, preferably mammals, such as non-human primates, sheep, dogs, cats, cows and horses.

[0065] "Effective dose" or "effective amount" refers to an amount sufficient to achieve or at least partially achieve the desired effect. A therapeutic "effective amount" or "effective dose" of a drug or therapeutic agent refers to an amount that, when used alone or in combination with another therapeutic agent, reduces the severity of disease symptoms, increases the frequency or duration of disease-free symptoms, or prevents or ameliorates damage caused by the disease or inhibits the development or recurrence of the disease.

[0066] "Cancer" refers to a collection of cells that proliferate in an abnormal manner and includes both solid and non-solid tumors.

[0067] Unless otherwise stated, "about" in the present invention means fluctuations within the range of ±5% of the given specific numerical range, preferably within the range of ±2%, and more preferably within the range of ±1%. For example, a pH value of about 5.5 means a pH of 5.5±5%, preferably a pH of 5.5±2%, and more preferably a pH of 5.5±1%. Unless otherwise specifically stated, the use of the singular includes the plural. Unless otherwise specifically stated, the word "one" or "a kind of" means "at least one" or "at least one", and the meaning of the phrase "at least one" is equivalent to the meaning of "one or more", and the use of "and / or" means "and" or "or".

[0068] Various aspects of the invention are described in more detail below.

[0069] Anti-PD-L2 monoclonal antibodies

[0070] Preferably, the antibodies of the present invention are antibodies whose structural and chemical characteristics are described below.

[0071] The anti-PD-L2 antibody or its antigen-binding fragment comprises a heavy chain CDR region and a light chain CDR region, wherein exemplary heavy chain CDR sequences and light chain CDR sequences are provided in Tables 1 and 2 below, respectively; the anti-PD-L2 antibody or its antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, wherein exemplary heavy chain variable regions and light chain variable regions are provided in Table 3 below. Some antibodies have the same CDR, and some antibodies have the same VH or VL. The heavy chain constant region of the antibody can be a human IgG1 heavy chain constant region, and the light chain constant region of the antibody can be a human kappa light chain constant region. These antibodies may also comprise a mouse IgG1 heavy chain constant region, or a human or mouse IgG2 or IgG4 heavy chain constant region, and / or a human or mouse lambda light chain constant region.

[0072] Table 1 Amino acid sequence of heavy chain CDR region

[0073]

[0074] Table 2 Amino acid sequence of light chain CDR region

[0075]

[0076] Table 3 Amino acid sequence table of heavy chain / light chain variable region (underlined CDR regions in sequence)

[0077]

[0078]

[0079] Table 4 Amino acid sequence of heavy chain / light chain (underline indicates variable region)

[0080]

[0081]

[0082]

[0083]

[0084] The VH and / or VL sequences (or CDR sequences) of other anti-PD-L2 antibodies that bind to human PD-L2 can be "mixed and paired" with the VH and / or VL sequences (or CDR sequences) of the antibodies of the present invention. Preferably, when the VH and VL chains (or their CDRs) are mixed and paired, the VH sequence in a specific VH / VL pair can be replaced by a structurally similar VH sequence. Similarly, the VL sequence in a specific VH / VL pair is preferably replaced with a structurally similar VL sequence.

[0085] Therefore, in one embodiment, the antibody or antigen-binding fragment thereof of the present invention comprises:

[0086] (a) a heavy chain variable region comprising the amino acid sequence listed in Table 3; and

[0087] (b) a light chain variable region comprising an amino acid sequence listed in Table 3, or a VL of another PD-L2 antibody, wherein the antibody specifically binds to human PD-L2.

[0088] In another embodiment, the antibody or antigen-binding fragment thereof of the invention comprises:

[0089] (a) the CDR1, CDR2 and CDR3 regions of the heavy chain variable region listed in Table 1; and

[0090] (b) the CDR1, CDR2 and CDR3 regions of the light chain variable region listed in Table 2, or the light chain variable region CDRs of another PD-L2 antibody, wherein the antibody specifically binds to human PD-L2.

[0091] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention comprises a heavy chain variable CDR2 region listed in Table 1 and CDRs of other PD-L2 antibodies, such as CDR1 and / or CDR3 of the heavy chain variable region of other PD-L2 antibodies, and / or CDR1, CDR2 and / or CDR3 of the light chain variable region of other PD-L2 antibodies.

[0092] In addition, it is well known in the art that the CDR3 domain, independent of the CDR1 and / or CDR2 domains, can independently determine the binding specificity of an antibody to the same antigen, and it can be predicted that multiple antibodies with the same binding specificity can be generated based on the CDR3 sequence.

[0093] In another embodiment, the antibody or antigen-binding fragment thereof of the present invention comprises a CDR2 of a heavy chain variable region listed in Table 1; and a CDR3 of a heavy chain variable region listed in Table 1 and / or a CDR3 of a light chain variable region listed in Table 2, or a CDR3 of a heavy chain variable region and / or a CDR3 of a light chain variable region of another PD-L2 antibody, wherein the antibody specifically binds to human PD-L2. These antibodies preferably (a) compete for binding to PD-L2; (b) retain functional characteristics; (c) bind to the same epitope; and / or (d) have a similar binding affinity to the PD-L2 antibody of the present invention. In another embodiment, the antibody of the present invention further comprises a CDR2 of a light chain variable region listed in Table 2, or a CDR2 of a light chain variable region of another PD-L2 antibody, wherein the antibody specifically binds to human PD-L2. In another embodiment, the antibody of the present invention further comprises a CDR1 of a heavy chain variable region listed in Table 1 and / or a CDR1 of a light chain variable region listed in Table 2, or a CDR1 of a heavy chain variable region and / or a CDR1 of a light chain variable region of another PD-L2 antibody, wherein the antibody specifically binds to human PD-L2.

[0094] Conservative Modifications

[0095] In another embodiment, the antibody of the present invention comprises a heavy chain variable region and / or a light chain variable region or CDR1, CDR2 and CDR3 sequence that has one or more conservative modifications with the PD-L2 antibody of the present invention. It should be understood in the art that some conservative sequence modifications will not eliminate antigen binding.

[0096] Thus, in one embodiment, the antibody comprises a heavy chain variable region and / or a light chain variable region, the heavy chain variable region and the light chain variable region respectively comprising CDR1, CDR2 and CDR3, wherein:

[0097] (a) the heavy chain variable region CDR1 sequence comprises the sequence listed in Table 1, and / or conservative modifications thereof; and / or

[0098] (b) the heavy chain variable region CDR2 sequence comprises the sequence listed in Table 1, and / or conservative modifications thereof; and / or

[0099] (c) the heavy chain variable region CDR3 sequence comprises the sequence listed in Table 1, and / or conservative modifications thereof; and / or

[0100] (d) the light chain variable region CDR1 and / or CDR2 and / or CDR3 sequences comprise the sequences listed in Table 2; and / or conservative modifications thereof; and

[0101] (e) The antibody specifically binds to human PD-L2.

[0102] The antibodies of the present invention have one or more of the following functional properties as described above, such as having high affinity for human PD-L2 and blocking the binding of PD-L2 / PD-1.

[0103] The term "conservative sequence modification" as used herein refers to amino acid modifications that do not significantly affect or change the binding properties of the antibody. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into the antibodies of the present invention by standard techniques known in the art, such as point mutations and PCR-mediated mutations. Conservative amino acid substitutions refer to amino acid residues that are replaced by amino acid residues with similar structures or chemical properties (e.g., similar side chains). Families of amino acid residues with similar side chains are known in the art. These amino acid residue families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), β-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues in the CDR region of an antibody of the invention may be substituted with other amino acid residues from the same side chain family, and the resulting antibodies may be tested for retention of function (ie, the function described above) using the functional assays described herein.

[0104] Genetically engineered antibodies and modified antibodies

[0105] The antibodies of the present invention can be prepared into genetically engineered antibodies using antibodies having one or more VH / VL sequences of the anti-PD-L2 antibodies of the present invention as starting materials. One or more residues in one or both variable regions (i.e., VH and / or VL) (e.g., in one or more CDR regions and / or one or more framework regions) can be modified. Alternatively, residues in the constant region can be modified, for example, to change the effector function of the antibody.

[0106] In certain embodiments, CDR transplantation can be used for the variable region of genetically engineered antibodies. Antibodies interact with target antigens primarily through amino acid residues located in six heavy and light chain CDRs. Therefore, the amino acid sequence in each antibody CDR is more diverse than the sequence outside the CDR. Since the CDR sequence is responsible for the main antibody-antigen interaction, it is possible to express a recombinant antibody that simulates the characteristics of a specific natural antibody by constructing an expression vector containing the CDR sequence of a specific natural antibody transplanted to the framework sequence of different antibodies of different characteristics.

[0107] Therefore, another embodiment of the present invention relates to a separated monoclonal antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises CDR1, CDR2 and CDR3 having the above-mentioned sequence of the present invention, and the light chain variable region comprises CDR1, CDR2 and CDR3 having the above-mentioned sequence of the present invention. Although these antibodies comprise the CDR sequences of the monoclonal antibodies of the present invention, they may contain different framework sequences. Such framework sequences may be obtained from public DNA databases or public references including germline antibody gene sequences. Such framework sequences are preferably those structurally similar to the framework sequences used by the antibodies of the present invention. CDR1, CDR2 and CDR3 sequences may be transplanted into a framework region having the same sequence as the germline immunoglobulin gene of the framework sequence, or CDR sequences may be transplanted into a framework region having one or more mutations compared to the germline sequence. For example, in some cases, it may be beneficial to mutate residues in the framework regions to maintain or enhance the antigen binding ability of the antibody (see, e.g., US Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370).

[0108] Another type of variable region modification is to mutate the amino acid residues in the CDR1, CDR2 and / or CDR3 regions to improve one or more properties (e.g., affinity, physicochemical properties) of the target antibody. Mutations can be introduced by fixed mutation or PCR-mediated mutation, and the effects of mutations on antibody binding or other functional properties can be assessed by in vitro or in vivo assays known in the art. Amino acids may be substituted, added or deleted, preferably substituted. In addition, no more than one, two, three, four or five residues in each CDR region are usually changed.

[0109] In addition, in another embodiment, the present invention provides an isolated PD-L2 monoclonal antibody or an antigen-binding fragment thereof, which comprises a heavy chain variable region and a light chain variable region, which comprises: (a) a VH CDR1 region comprising a sequence of the present invention, or an amino acid sequence having 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions; (b) a VH CDR2 region comprising a sequence of the present invention, or an amino acid sequence having 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions; (c) a VH CDR3 region comprising a sequence of the present invention, or an amino acid sequence having 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions; (d) a VL CDR1 region comprising a sequence of the present invention, or an amino acid sequence having 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions; (e) a VL CDR2 region comprising a sequence of the present invention, or an amino acid sequence having 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions; and (f) a VL A CDR3 region comprising a sequence of the present invention, or an amino acid sequence having 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions.

[0110] Modified antibodies of the present invention include those in which the framework region residues of VH and / or VL are modified to improve the antibody properties. Generally, such framework region modifications can reduce the immunogenicity of the antibody. For example, one or more framework residues are "back mutated" to the corresponding germline sequence. These residues can be identified by comparing the antibody framework sequence with the germline sequence of the resulting antibody.

[0111] Another type of framework modification involves mutating one or more residues in the framework region, or even one or more CDR regions, to remove T cell epitopes, thereby reducing the immunogenicity that the antibody may cause. This method is also called "de-immunization" and is described in more detail in U.S. Patent Publication No. 20030153043.

[0112] In addition, the antibody Fc region of the present invention is also modified, which is generally used to change one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular toxicity. In addition, the antibody of the present invention can also be chemically modified (e.g., connected to one or more chemical functional groups), or modified to change its glycosylation, to change one or more functional properties of the antibody. In other embodiments, the Fc region is modified by pegylation (e.g., by reacting the antibody or its fragment with polyethylene glycol (PEG).

[0113] Pharmaceutical composition

[0114] In another aspect, the present invention provides a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof of the present invention, formulated together with a pharmaceutically acceptable carrier. The pharmaceutical composition may optionally comprise one or more other pharmaceutically effective ingredients, such as another antibody or drug, such as another anti-PD-L2 antibody, anti-PD-L1 antibody, anti-PD-1 antibody, another anticancer drug, preferably comprising another anti-PD-L1 antibody.

[0115] As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antimicrobial agents, isotonic agents and combinations thereof that are physiologically compatible. The selection and use of suitable "pharmaceutically acceptable carriers" are taught in Gennaro, ed., Remington: The Science and Practice of Pharmacy, 20th Ed. (Lippincott Williams & Wilkins 2003).

[0116] In some embodiments, the pharmaceutical composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Based on the difference in the route of administration, the active ingredient can be wrapped in a material to protect it from acid and other natural conditions that may inactivate it. "Enteral administration" refers to a method different from intestinal and topical administration, usually by injection, including but not limited to intravenous, intramuscular, intraarterial, intramembranous, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, supradinaline and intrasternal injection and push injection. Alternatively, the antibody of the present invention or its antigen-binding fragment can be administered by a non-parenteral route, such as topical, epidermal or mucosal administration, such as intranasal, oral, vaginal, rectal, sublingual, or topical administration.

[0117] Various delivery systems are known and can be used to administer one or more antibodies or antigen-binding fragments thereof of the present invention, or a combination of one or more antibodies or antigen-binding fragments thereof of the present invention and one or more other pharmaceutically effective ingredients, such as another antibody or drug, e.g., another PD-L2 antibody, an anti-PD-L1 antibody, another anti-cancer drug.

[0118] Uses and methods of the invention

[0119] The antibodies or antigen-binding fragments thereof of the present invention (nucleic acids encoding the antibodies or antigen-binding fragments thereof of the present invention, pharmaceutical compositions, recombinant polypeptides, fusion proteins or bispecific molecules) have a variety of in vitro and in vivo applications, such as for regulating immune responses in subjects, enhancing anti-tumor immune responses, reducing tumor size or inhibiting tumor cell growth, and / or treating (e.g., curing, inhibiting, improving, delaying or preventing recurrence) tumors or cancers. Therefore, the present invention also provides a method for enhancing anti-tumor immune responses in subjects, comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof. In one embodiment, the present invention also provides a method for reducing tumor size or inhibiting tumor cell growth in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof. In one embodiment, the present invention also provides a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof. In other embodiments, the method comprises administering to the subject a therapeutically effective amount of a nucleic acid encoding an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention.

[0120] In other embodiments, the method comprises administering to the subject a therapeutically effective amount of a recombinant polypeptide, fusion protein or bispecific molecule of the invention. The anti-PD-L2 antibody or antigen-binding fragment thereof described herein can be administered to a human subject alone or in combination with one or more other antibodies, such as an anti-PD-L1 antibody, an anti-PD-1 antibody and / or an anti-CTLA-4 antibody, preferably an anti-PD-L1 antibody. The anti-PD-L2 antibody or antigen-binding fragment thereof described herein can be administered to a human subject alone or in combination with at least one other form of treatment, including radiotherapy, chemotherapy, biological products, chemicals and vaccines.

[0121] Tumors or cancers discussed herein include, but are not limited to, colorectal cancer, breast cancer, ovarian cancer, pancreatic cancer, gastric cancer, renal cancer, prostate cancer, cervical cancer, myeloma, lymphoma, leukemia, thyroid cancer, endometrial cancer, uterine cancer, bladder cancer, neuroendocrine malignancies, head and neck cancer, salivary gland cancer, thymic cancer, liver cancer, nasopharyngeal cancer, testicular cancer, small cell lung cancer, non-small cell lung cancer, melanoma, basal cell skin cancer, squamous cell skin cancer, dermatofibrosarcoma protuberans, Merkel cell carcinoma, glioblastoma, glioma, sarcoma, mesothelioma, and myelodysplastic syndrome, or a combination thereof; and optionally, the tumor is in situ, metastatic, recurrent, and / or refractory.

[0122] In another embodiment, the present invention also provides a method for treating an infectious disease in a subject, the method comprising administering to the subject a therapeutically effective amount of an anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention or a pharmaceutical composition thereof. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a nucleic acid encoding an antibody or antigen-binding fragment thereof of the present invention. The infectious diseases include, but are not limited to, candidiasis, candidemia, aspergillosis, streptococcal pneumonia, streptococcal skin and oropharyngeal disease, gram-negative sepsis, tuberculosis, mononucleosis, influenza, respiratory diseases caused by respiratory syncytial virus, malaria, schistosomiasis, and trypanosomiasis.

[0123] A "combination" or "combination" of therapeutic agents discussed herein can be administered simultaneously as a single composition in a pharmaceutically acceptable carrier, or as separate compositions, wherein each agent is in a pharmaceutically acceptable carrier. In another embodiment, the combination of therapeutic agents can be administered sequentially. In addition, if multiple administrations of the combination therapy are performed, and the agents are administered sequentially, the order of sequential administration at each time point can be reversed or remain the same, and sequential administration can be combined with simultaneous administration or any combination thereof.

[0124] On the other hand, the present invention also provides a method for detecting PD-L2 expression in a sample, comprising contacting the sample with the PD-L2 antibody or antigen-binding fragment thereof of the present invention under conditions allowing formation of a complex between the anti-PD-L2 antibody or antigen-binding fragment thereof of the present invention and PD-L2, and detecting the formation of the complex.

[0125] Although the foregoing invention has been described in considerable detail by way of illustration and examples for the purpose of clear understanding, it will be apparent to one of ordinary skill in the art that certain changes and modifications may be made to the invention in addition without departing from the spirit and scope of the appended claims, based on the teachings of the present invention. The present invention is further illustrated by the following examples, which are not intended to be limiting. One of ordinary skill in the art will readily recognize a variety of noncritical parameters that may be changed or modified to produce substantially similar results.

[0126] The practice of the present invention will employ, unless otherwise indicated, conventional methods of protein chemistry, biochemistry, recombinant DNA techniques and pharmacology, which are within the skill of the art. Example

[0127] Example 1: Generation of anti-PD-L2 monoclonal antibodies

[0128] To produce antibodies against human PD-L2, an equal volume of recombinant hPD-L2-His protein (ACRObiosystems, catalog number: PD2-H5220) emulsified with complete Freund's adjuvant was used for the first immunization, 500 μg protein / mouse. An equal volume of recombinant hPD-L2-mFc protein (ACRObiosystems, catalog number: PD2-H52A5) emulsified with incomplete Freund's adjuvant was used for booster immunization, 500 μg protein / mouse, and Balb / c mice were subcutaneously immunized every 2 weeks for 6 weeks. The fourth immunization was performed by intramuscular injection of hPDL2-mFc protein (20 μg / mouse). Three days before fusion, immunization was boosted by subcutaneous injection of antigen without adjuvant. Splenocytes (1×10 8 ) and SP2 / 0 myeloma cells (1.5×10 7 ) fusion. After fusion, the cells were distributed into 96-well plates at 0.1 ml / well and incubated in an incubator at 37°C, 5% CO2. Usually between the 9th and 15th days of feeding, the binding of the culture supernatant to hPD-L2-His (ACRObiosystems, catalog number: PD2-H5220) was detected by ELISA. The operation steps can be found in Example 4.

[0129] The screened mixed clones that can bind to hPD-L2 were subjected to limiting dilution, and one positive mixed clone was distributed to multiple 96-well plates and cultured in RPMI 1640 (Hyclone, catalog number: SH30809.01) containing 10% fetal bovine serum. After 10 days, the subclones after limiting dilution were also subjected to ELISA experiments to detect the binding of the culture supernatant to hPD-L2-His (ACRObiosystems, catalog number: PD2-H5220), and then the positive subclones that can bind to hPD-L2 were picked.

[0130] Example 2: Anti-PD-L2 antibody cDNA sequence cloning and humanization

[0131] Cloning of immunoglobulin cDNA

[0132] Total RNA isolated from a hybridoma cell line producing human PD-L2 antibodies was used as a template using an RNA extraction kit (Takara, catalog number: 9767), and the first-strand cDNA was synthesized using a reverse transcription kit (Thermo Fisher, catalog number: K1652) according to the manufacturer's instructions. Antibody-related sequences were then amplified in vitro by PCR using degenerate mouse IgG primers in a 50 ml reaction volume. PCR reactions were performed in a T100TM thermal cycler (Bio-Rad, catalog number: 184-2000).

[0133] The PCR mixture was electrophoresed in a 1% agarose / Tris-acetate gel, and a DNA fragment of the expected size was cut from the gel and purified. The purified PCR product was cloned into a pMD-19T vector (Takara, catalog number: 6013), and then transformed into DH5a competent E. coli cells (Takara, catalog number: 9057). The clones were screened by colony PCR, and the clones were selected for DNA sequencing. The heavy chain variable region sequences and light chain variable region sequences of the antibodies were obtained: 25G4 (SEQ ID NO: 25-26), 14E7 (SEQ ID NO: 27-28), 24A3 (SEQ ID NO: 29-30), 7A7 (SEQ ID NO: 31-32), as shown in Table 3.

[0134] Construction and expression of chimeric 7A7, 14E7, 24A3 and 25G4 antibodies

[0135] The genes of the mouse variable regions were chemically synthesized, wherein the cDNA of the VL region was connected to the human κ light chain constant region to construct the chimeric light chains of 25G4 (SEQ ID NOs: 47, 49), 14E7 (SEQ ID NOs: 51, 53), 24A3 (SEQ ID NOs: 55, 57) and 7A7 (SEQ ID NOs: 59, 61); the cDNA of the VH region was connected to the human IgG1 constant region to construct the chimeric heavy chains of 25G4 (SEQ ID NOs: 46, 48), 14E7 (SEQ ID NOs: 50, 52), 24A3 (SEQ ID NOs: 54, 56) and 7A7 (SEQ ID NOs: 58, 60).

[0136] 100 μg of each chimeric heavy chain expression plasmid and chimeric light chain expression plasmid were transfected into Expi-CHO cells (200 mL, density 10 6 / mL) and cultured for 6 days. The chimeric antibody in the supernatant was then purified using a protein A column (Genscript, catalog number: L00695-16). "Xi" in the present invention represents a chimeric antibody, for example, Xi7A7 represents a chimeric 7A7 antibody.

[0137] Humanized antibody design

[0138] Antibody humanization was performed using the CDR transplantation method (see, e.g., U.S. Pat. No. 5,225,539). The VH and VL amino acid sequences of mouse 25G4, 14E7, 24A3 and 7A7 antibodies were respectively input into the website of the International Immunogenetic Information System Related Tools (http: / / www.imgt.org / 3Dstructure-DB / cgi / DomainGapAlign.cgi), and human germline antibody sequences with high homology to mouse 25G4, 14E7, 24A3 and 7A7 antibodies were screened out as human antibody templates for grafting complementary determining regions (CDRs) in VH and VL of mouse 25G4, 14E7, 24A3 and 7A7 antibodies. For 25G4, the selected human VH template was a combination of IGHV1-46*01 and IGHJ4*01, and the selected human VL template was a combination of IGKV1-39*01 and IGKJ1*01. For 14E7, the selected human VH template is a combination of IGHV1-46*01 and IGHJ4*01, and the selected human VL template is a combination of IGKV1-5*01 and IGKJ1*01. For 24A3, the selected human VH template is a combination of IGHV1-46*01 and IGHJ4*01, and the selected human VL template is a combination of IGKV4-1*01 and IGKJ1*01. For 7A7, the selected human VH template is a combination of IGHV1-46*01 and IGHJ4*01, and the selected human VL template is a combination of IGKV1-39*01 and IGKJ1*01.

[0139] The CDR amino acid sequences of the above human antibody templates are replaced by the CDRs of mouse 25G4 (SEQ ID NO: 1-6), 14E7 (SEQ ID NO: 7-12), 24A3 (SEQ ID NO: 13-18) and 7A7 (SEQ ID NO: 19-24) antibodies, and the essential amino acid sequences of VH and VL of mouse 25G4, 14E7, 24A3 and 7A7 antibodies are grafted into the above human germline antibody templates VH and VL to obtain functional humanized antibodies. For VH and VL of 25G4, 14E7, 24A3 and 7A7 antibodies, several sites of the framework amino acids of the above human antibody templates are backmutated to the corresponding amino acid sequences in mouse 25G4, 14E7, 24A3 and 7A7 antibodies. Some CDRs are mutated on this basis to improve the properties of the antibody, such as improving the physicochemical properties or drugability. Humanized antibodies Hz25G4-1.1, Hz25G4-1.2, Hz14E7-1.1, Hz14E7-1.2, Hz24A3-1.1, Hz24A3-1.2, Hz7A7-1.1, Hz7A7-1.2, Hz7A7-2.1, and Hz7A7-2.2 were obtained, wherein the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz25G4-1.1 are SEQ ID NO: 33 and SEQ ID NO: 34, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz25G4-1.2 are SEQ ID NO: 35 and SEQ ID NO: 34, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz14E7-1.1 are SEQ ID NO: 36 and SEQ ID NO: 37, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz14E7-1.2 are SEQ ID NO: 38 and SEQ ID NO: 39, respectively. NO: 37; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz24A3-1.1 are SEQ ID NO: 39 and SEQ ID NO: 40, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz24A3-1.2 are SEQ ID NO: 41 and SEQ ID NO: 40, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz7A7-1.1 are SEQ ID NO: 42 and SEQ ID NO: 43, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz7A7-1.2 are SEQ ID NO: 44 and SEQ ID NO: 43, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz7A7-2.1 are SEQ ID NO: 42 and SEQ ID NO: 45, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of Hz7A7-2.2 are SEQ ID NO: 44 and SEQ ID NO: 45, respectively.

[0140] Construction and expression of humanized 25G4, 14E7, 24A3 and 7A7 antibodies

[0141] DNA encoding humanized 25G4, 14E7, 24A3 and 7A7 antibody VL and VH were synthesized, and the gene synthesis fragments of the humanized VL region were connected to the human κ light chain constant region by double enzyme digestion to construct the light chains of humanized 25G4-1.1 and 25G4-1.2 (SEQ ID NOs: 63, 67), 14E7-1.1 and 14E7-1.2 (SEQ ID NOs: 69, 73), 24A3-1.1 and 24A3-1.2 (SEQ ID NOs: 75, 79), 7A7-1.1 and 7A7-1.2 (SEQ ID NOs: 81, 86), 7A7-2.1 and 7A7-2.2 (SEQ ID NOs: 83, 87); the gene synthesis fragments of the humanized VH region were connected to the human IgG1 constant region by double enzyme digestion to construct the humanized 25G4-1.1 (SEQ ID NOs: 81, 86). NO: 62, 65), 25G4-1.2 (SEQ ID NO: 64, 66), 14E7-1.1 (SEQ ID NO: 68, 71), 14E7-1.2 (SEQ ID NO: 70, 72), 24A3-1.1 (SEQ ID NO: 74, 77), 24A3-1.2 (SEQ ID NO: 76, 78), 7A7-1.1 and 7A7-2.1 (SEQ ID NO: 80, 84), 7A7-1.2 and 7A7-2.2 (SEQ ID NO: 82, 85) heavy chains.

[0142] 100 μg of each humanized heavy chain expression plasmid and humanized light chain expression plasmid were transfected into Expi-CHO cells (200 mL, density 10 6 / mL), and cultured for 6 days. Then, the humanized antibody in the supernatant was purified using a protein A column (Genscript, catalog number: L00695-16). "Hz" in the present invention means a humanized antibody, for example, Hz7A7-1.1 means a humanized 7A7-1.1 antibody.

[0143] Example 3: Binding affinity analysis of anti-PD-L2 antibodies to PD-L2

[0144] The binding kinetics of anti-PD-L2 antibody to PD-L2 was detected by Biacore T200 (GE). The experimental steps were as follows: Anti-human IgG antibody was connected to the reference flow cell and experimental flow cell of the CM5 chip by amino coupling. At the beginning of each cycle, the anti-PD-L2 antibody was injected into the experimental flow cell to be captured. The PD-L2 protein was then diluted with running buffer and continuously injected into the reference cell and the experimental cell. The reaction signal was detected in real time by Biacore to obtain the binding and dissociation curves. Finally, the experimental data were analyzed using BiaEvaluation Software 2.0 software with a 1:1 (Langmuir) binding model to determine the association rate constant ka (kon) and the dissociation rate constant kd (koff), and the dissociation constant KD was calculated by KD = kd / ka. The affinity data of anti-PD-L2 antibodies to human PD-L2 antigen (hPD-L2-His: ACRObiosystems, catalog number: PD2-H5220) and cynomolgus monkey PD-L2 antigen (Cyno PD-L2-His: Sino Biological, catalog number: 90249-C08H) are shown in Tables 5 and 6.

[0145] Table 5 Binding affinity of anti-PD-L2 chimeric antibodies to PD-L2

[0146]

[0147] Table 6 Binding affinity of anti-PD-L2 humanized antibodies to PD-L2

[0148]

[0149] NA: Not reaching the detection line

[0150] Example 4: ELISA-based binding analysis of anti-PD-L2 antibodies

[0151] Based on the ELISA method, PD-L2-His was used as an antigen to analyze the binding ability of anti-PD-L2 antibodies to PD-L2. The experimental steps were as follows: PBS buffer containing 2 μg / ml hPD-L2-His (ACRObiosystems, catalog number: PD2-H5220) was coated in a 96-well plate at 100 μl / well and incubated overnight at 4°C. The liquid in the 96-well plate was removed, and 100 μL of blocking buffer (PBS buffer containing 1% (w / v) bovine serum albumin) was added to each well and incubated at 37°C for 1 hour. After washing the 96-well plate three times with washing buffer (PBS buffer containing 0.01% (w / v) Tween 20), 100 μl / well of anti-PD-L2 antibody (initial concentration is 30 μg / mL, serially diluted according to 1:4; that is, the concentration is about 30 μg / mL, about 7.5 μg / mL, about 1.875 μg / mL, about 0.469 μg / mL, about 0.117 μg / mL, about 0.029 μg / mL, etc.) was added and incubated at room temperature for 2 hours. After washing the 96-well plate three times with washing buffer, 100 μl / well of blocking buffer containing biotin (Thermo, catalog number: 21336) was added and labeled at room temperature for 1 hour. After washing the 96-well plate three times with washing buffer, the second antibody Avidin-HRP was added and incubated in the dark for 20 minutes. After washing the 96-well plate 3 times, 100 μl / well of substrate solution TMB (TIANGEN, catalog number: PA107-01) was added, and the 96-well plate was incubated at room temperature for 10 minutes, and then 100 μl / well of stop solution (1M H2SO4) was added to stop the reaction. The colorimetric signal generated was read at 450 nm using a multifunctional microplate reader (PerkinElmer, EnVision), and the experimental data was analyzed using GraphPadPrism5, and the EC50 was calculated. The EC50 of the binding of anti-PD-L2 humanized antibodies to PD-L2 in the ELISA test is shown in Table 7.

[0152] Table 7 EC50 of binding of anti-PD-L2 humanized antibodies to PD-L2 in ELISA assay

[0153] Antibody Hz7A7-1.1 Hz7A7-1.2 Hz7A7-2.1 Hz7A7-2.2 Hz14E7-1.1 EC50(ng / mL) 308.2 679.6 1125 NA 115.4 Antibody Hz14E7-1.2 Hz24A3-1.1 Hz24A3-1.2 Hz25G4-1.1 Hz25G4-1.2 EC50(ng / mL) 33.28 166.5 223 68.93 87.4

[0154] NA: Not reaching the detection line

[0155] Example 5: Flow cytometry-based cell binding analysis of anti-PD-L2 antibodies

[0156] The binding ability of anti-PD-L2 antibodies to cells expressing PD-L2 was analyzed by binding experiments with the U2OS cell line stably expressing human PD-L2 (PD-L2-U2OS). 3U2OS-PD-L2 cells were added to each well of a 96-well U-shaped plate (Corstar, catalog number: 3799), and anti-PD-L2 antibody (initial concentration of 50 μg / ml, serially diluted 1:5) was added to the cell suspension and incubated at room temperature for 1 to 2 hours. After washing the cells three times with PBS buffer, Alexa Fluor 488-labeled goat anti-human secondary antibody (Jackson ImmunoResearch Inc, catalog number: 109-545-088) diluted 1:200 was added at 100 μl / well and incubated at room temperature for 30 minutes. After washing with PBS three times, the fluorescence signal was analyzed and detected by flow cytometry (BD, Accuri C6). The binding ability of anti-PD-L2 antibody to PD-L2 on PD-L2-U2OS cells was measured by the mean fluorescence intensity (MFI) of staining. The data were analyzed using GraphPad Prism5, and the EC50 value was calculated. The EC50s of the binding of anti-PD-L2 chimeric antibodies and anti-PD-L2 humanized antibodies to PD-L2 based on flow cytometry are shown in Tables 8 and 9, respectively.

[0157] Table 8 EC50 of anti-PD-L2 chimeric antibodies binding to PD-L2 cells based on flow cytometry

[0158] Antibody Xi7A7 Xi14E7 Xi24A3 Xi25G4 EC50(ng / mL) 429.6 1125 967 540

[0159] Table 9 EC50 of anti-PD-L2 humanized antibodies binding to PD-L2 based on flow cytometry

[0160] Antibody Hz7A7-1.1 Hz7A7-1.2 Hz7A7-2.1 Hz7A7-2.2 Hz14E7-1.1 EC50(ng / mL) 613.1 832.7 924.3 1735 442.8 Antibody Hz14E7-1.2 Hz24A3-1.1 Hz24A3-1.2 Hz25G4-1.1 Hz25G4-1.2 EC50(ng / mL) NA 1048 619.8 482.8 NA

[0161] NA: Not reaching the detection line

[0162] Example 6: Cell-based antibody blocking assay for anti-PD-L2 antibodies

[0163] PD-L2 is the ligand of PD-1, and anti-PD-L2 antibodies can block the binding of PD-L2 to its receptor PD-1. Take receptor cells that are in good exponential growth phase (PD-1-Jurkat suspension cells that stably express human PD-1) and adjust the cell density to 6.4x10 5 / mL, inoculated on a 384-well plate at 12.5μL / well, and cultured in a 37°C, 5% CO2 incubator for 15 minutes. Add 2.5μL / well of anti-PD-L2 antibody (initial concentration of 50μg / ml, serially diluted at 1:5), set to 37°C, 5% CO2 incubator and incubate for 60 minutes. Take the ligand cells (U2OS cell line stably expressing human PD-L2, PD-L2-U2OS) in the exponential growth phase and adjust the cell density to 9.6x10 5 / mL, inoculated in the above 384-well plate at 12.5μL / well, and incubated on a shaker at room temperature for 2 hours. Add 2.5μL Bioassay Reagent (DiscoverX, catalog number: 12604-021) to each well, incubate on a shaker at room temperature in the dark for 15 minutes, then add 10μL Bioassay Reagent 2 (DiscoverX, catalog number: 12604-021) to each well, and incubate on a shaker at room temperature in the dark for 60 minutes. The chemiluminescent signal was read by a multifunctional microplate reader EnVision (PerkinElmer, MNR0643,), and the data was analyzed using GraphPad Prism5, and the IC50 value was calculated. The IC50 of the anti-PD-L2 chimeric antibody blocking the binding of PD-L2 to PD-1 is shown in Table 10, and the IC50 of the anti-PD-L2 humanized antibody blocking the binding of PD-L2 to PD-1 is shown in Table 11.

[0164] Table 10 IC50 of anti-PD-L2 chimeric antibodies blocking the binding of PD-L2 and PD-1

[0165] Antibody Xi7A7 Xi14E7 Xi24A3 Xi25G4 IC50(ng / mL) NA 342 220.8 679.8

[0166] Table 11 IC50 of anti-PD-L2 humanized antibodies blocking the binding of PD-L2 and PD-1

[0167] Antibody Hz7A7-1.1 Hz7A7-1.2 Hz7A7-2.1 Hz7A7-2.2 Hz14E7-1.1 IC50(ng / mL) 585.4 373.5 144.3 NA 111.7 Antibody Hz14E7-1.2 Hz24A3-1.1 Hz24A3-1.2 Hz25G4-1.1 Hz25G4-1.2 IC50(ng / mL) 2663 1953 1423 1005 700.4

[0168] NA: Not reaching the detection line

[0169] Example 7: Binding of anti-PD-L2 humanized antibodies to DC / T cells

[0170] FACS was used to analyze the binding of anti-PD-L2 antibody to DC / T cells, using CD4 + T cell isolation kit (Mitenyi Biotech, catalog number: 130-094-131) was used to isolate human CD4 T cells from human PBMCs. + T cells. Immature dendritic cells (DCs) were derived from monocytes isolated from human PBMCs using a human monocyte isolation kit (Stemcell, catalog number: 19319), and the cells were cultured with a dendritic cell differentiation medium (Stemcell, catalog number: 10985) for 5 days, and then induced with a dendritic cell maturation medium (Stemcell, catalog number: 10985) for 2 days to obtain mature DCs. Mature DCs were separated from CD4 +T cells were mixed at a ratio of 1:4 in terms of cell number, and then anti-PD-L2 antibodies at concentrations of 4 μg / mL, 0.8 μg / mL, 0.16 μg / mL, 0.032 μg / mL, and 0.0064 μg / mL were added. After incubation at 37°C and 5% CO2 incubator for 96 hours, the supernatant was collected, and goat anti-human 488-labeled fluorescent secondary antibodies diluted 1:200 were added to the cells. The fluorescent signals were analyzed by flow cytometry (BD, Accuri C6) to detect the binding of antibodies to immune checkpoints on cells. The results are shown in Figure 1 As shown, the anti-PD-L2 antibody Hz25G4-1.1 can bind to immune checkpoints on cells.

[0171] Example 8: Effect of anti-PD-L2 humanized antibody on T cell activation in mixed lymphocyte reaction

[0172] Mixed lymphocyte reaction (MLR) was used to demonstrate the effect of anti-PD-L2 humanized antibody, or anti-PD-L2 humanized antibody combined with PD-L1 antibody, in blocking the binding of PD-1 to its ligand in lymphocyte effector cells, and to measure the secretion of IFN-γ and IL-2 by T cells. + T cell isolation kit (Mitenyi Biotech, catalog number: 130-094-131) was used to purify human CD4 T cells from human PBMCs. + T cells. Immature dendritic cells (DCs) were derived from monocytes isolated from human PBMCs using a human monocyte isolation kit (Stemcell, catalog number: 19319). The cells were cultured for 5 days with a dendritic cell differentiation medium (Stemcell, catalog number: 10985), and then induced for 2 days with a dendritic cell maturation medium (Stemcell, catalog number: 10985) to obtain mature DCs. To establish an MLR, 10 5 Purified CD4 + T cells and 10 4Allogeneic mature DC cells with a total volume of 200 μL. Experimental samples were added separately, and the cells were cultured in an incubator at a constant temperature of 37°C and 5% CO2 for 96 hours. The experimental samples were divided into: (1) Hz25G4-1.1 + anti-PD-L1 antibody (wherein the concentrations of Hz25G4-1.1 were 4 μg / mL, 0.8 μg / mL, 0.16 μg / mL, 0.032 μg / mL, 0.0064 μg / mL, and the concentration of anti-PD-L1 antibody was 0.16 μg / mL); (2) anti-PD-L1 antibody + Hz25G4-1.1 (wherein the concentrations of anti-PD-L1 antibody were 4 μg / mL, 0.8 μg / mL, 0.16 μg / mL, 0.032 μg / mL, 0.0064 μg / mL, and the concentration of Hz25G4-1 .1 concentration was 4 μg / mL); (3) Hz25G4-1.1 (concentrations were 4 μg / mL, 0.8 μg / mL, 0.16 μg / mL, 0.032 μg / mL, 0.0064 μg / mL); (4) anti-PD-L1 antibody (concentrations were 4 μg / mL, 0.8 μg / mL, 0.16 μg / mL, 0.032 μg / mL, 0.0064 μg / mL); (5) blank control IgG1 (concentrations were 4 μg / mL, 0.8 μg / mL, 0.16 μg / mL, 0.032 μg / mL, 0.0064 μg / mL). The levels of IFN-γ and IL-2 in the culture medium were measured using an IL-2 ELISA detection kit (R&D, catalog number: DY202-05) and a hIFN-γ detection ELISA kit (R&D, catalog number: DY285). The PD-L1 antibody in this example refers to the humanized 5G11-IgG1 in CN107001463, and the amino acid sequences of the heavy chain and light chain are shown in SEQ ID NO: 88 and SEQ ID NO: 89 of the present application. Figure 2a and 2b As shown, the anti-PD-L2 humanized antibody Hz25G4-1.1 combined with the PD-L1 antibody promoted T cells to secrete IFN-γ and IL-2, showing a synergistic effect with obvious dose dependence.

[0173] Example 9: Detection of thermal stability of anti-PD-L2 antibodies based on NanoDSF

[0174] The high-throughput protein stability analyzer Prometheus NT.48 (NanoTemper) is a device based on differential scanning fluorescence (nanoDSF) technology to detect various types of data such as protein molecular structure stability, aggregation stability, and colloidal dispersion stability. The device was used to detect the melting temperature (Tm) and aggregation temperature (Tagg) of anti-PD-L2 chimeric antibodies and anti-PD-L2 humanized antibodies. The results are shown in Tables 12 and 13, indicating that the anti-PD-L2 antibody has good thermal stability.

[0175] Table 12 Thermal stability of anti-PD-L2 chimeric antibodies

[0176] Antibody Xi7A7 Xi14E7 Xi24A3 Xi25G4 Tm(℃) 65.6 65.61 60.71 65.85 Tagg(℃) 66.94 67.76 61.26 67.53

[0177] Table 13 Thermal stability of anti-PD-L2 humanized antibodies

[0178] Antibody Hz7A7-1.1 Hz7A7-1.2 Hz7A7-2.1 Hz7A7-2.2 Hz14E7-1.1 Tm(℃) 69.22 68.45 68.59 68.48 65.89 Tagg(℃) 70.09 69.16 69.26 69.14 84.11 Antibody Hz14E7-1.2 Hz24A3-1.1 Hz24A3-1.2 Hz25G4-1.1 Hz25G4-1.2 Tm(℃) 66.77 66.29 66.02 69.83 69.78 Tagg(℃) 83.82 79.65 76.91 70.86 71.20

[0179] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention. Sequence Listing <110> Chia Tai Tianqing Pharmaceutical Group Co., Ltd. <120> Anti-PD-L2 antibodies <150> CN202010833373.2 <151> 2020-08-18 <160> 89 <170> SIPOSequenceListing 1.0 <210> 1 <211> 10 <212> PRT <213> Mouse (Mus sp.) <400> 1 Gly Tyr Thr Phe Thr Thr Asn Trp Met Asn 1 5 10 <210> 2 <211> 10 <212> PRT <213> Mouse (Mus sp.) <220> <221> UNSURE <222> (6) <223> Xaa can be D or E <400> 2 Arg Ile Asp Pro Ser Xaa Ser Glu Thr His 1 5 10 <210> 3 <211> 10 <212> PRT <213> Mouse (Mus sp.) <400> 3 Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr 1 5 10 <210> 4 <211> 15 <212> PRT <213> Mouse (Mus sp.) <400> 4 Arg Ala Ser Glu Ser Val Asp Asn Tyr Gly Ile Ser Leu Met His 1 5 10 15 <210> 5 <211> 7 <212> PRT <213> Mouse (Mus sp.) <400> 5 Arg Ala Ser Asp Leu Glu Ser 1 5 <210> 6 <211> 9 <212> PRT <213> Mouse (Mus sp.) <400> 6 Gln Gln Ser Asp Lys Asp Pro Pro Thr 1 5 <210> 7 <211> 10 <212> PRT <213> Mouse (Mus sp.) <400> 7 Gly Tyr Ser Phe Thr Ser Tyr Trp Met His 1 5 10 <210> 8 <211> 10 <212> PRT <213> Mouse (Mus sp.) <220> <221> UNSURE <222> (6) <223> Xaa can be D or E <400> 8 Met Ile Asp Pro Ser Xaa Ser Glu Thr Arg 1 5 10 <210> 9 <211> 8 <212> PRT <213> Mouse (Mus sp.) <400> 9 Trp Ala Leu Gln Tyr Phe Asp Tyr 1 5 <210> 10 <211> 11 <212> PRT <213> Mouse (Mus sp.) <400> 10 Lys Ala Ser Gln Asn Val Gly Ala Ala Val Ala 1 5 10 <210> 11 <211> 7 <212> PRT <213> Mouse (Mus sp.) <400> 11 Ser Ala Ser Asn Arg Tyr Pro 1 5 <210> 12 <211> 9 <212> PRT <213> Mouse (Mus sp.) <400> 12 Gln Gln Tyr Arg Pro Tyr Pro Phe Thr 1 5 <210> 13 <211> 10 <212> PRT <213> Mouse (Mus sp.) <400> 13 Gly Tyr Ala Phe Thr Asn Tyr Leu Ile Glu 1 5 10 <210> 14 <211> 10 <212> PRT <213> Mouse (Mus sp.) <220> <221> UNSURE <222> (6) <223> Xaa can be N or Q <400> 14 Val Ile Asn Pro Val Xaa Gly Gly Ile Ser 1 5 10 <210> 15 <211> 9 <212> PRT <213> Mouse (Mus sp.) <400> 15 Ser Gly Gly Asn Tyr Gly Gly Asp Tyr 1 5 <210> 16 <211> 11 <212> PRT <213> Mouse (Mus sp.) <400> 16 Lys Ala Ser Gln Ser Val Ser Asn Glu Val Ala 1 5 10 <210> 17 <211> 7 <212> PRT <213> Mouse (Mus sp.) <400> 17 Tyr Ala Ser His Arg Tyr Thr 1 5 <210> 18 <211> 9 <212> PRT <213> Mouse (Mus sp.) <400> 18 Gln Gln His Phe Ser Ser Pro Leu Thr 1 5 <210> 19 <211> 10 <212> PRT <213> Mouse (Mus sp.) <400> 19 Gly Tyr Ser Phe Ser Gly Tyr Thr Met Asn 1 5 10 <210> 20 <211> 10 <212> PRT <213> Mouse (Mus sp.) <220> <221> UNSURE <222> (6) <223> Xaa can be N or Q <400> 20 Leu Ile Ser Pro Tyr Xaa Gly Val Thr Thr 1 5 10 <210> twenty one <211> 10 <212> PRT <213> Mouse (Mus sp.) <400> twenty one Gly Val Thr Val Pro Leu Arg Phe Asp Val 1 5 10 <210> twenty two <211> 17 <212> PRT <213> Mouse (Mus sp.) <220> <221> UNSURE <222> (8) <223> Xaa can be N or Q <400> twenty two Lys Ser Ser Gln Ser Leu Leu Xaa Ser Gly Asn Gln Lys Asn Tyr Leu 1 5 10 15 Thr <210> twenty three <211> 7 <212> PRT <213> Mouse (Mus sp.) <400> twenty three Trp Ala Ser Thr Arg Glu Ser 1 5 <210> twenty four <211> 9 <212> PRT <213> Mouse (Mus sp.) <400> twenty four Gln Asn Asp Asp Ser Tyr Pro Leu Thr 1 5 <210> 25 <211> 119 <212> PRT <213> Mouse (Mus sp.) <400> 25 Gln Val Gln Leu Gln Gln Pro Gly Val Glu Val Val Arg Pro Gly Ala 1 5 10 15 Pro Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Asn 20 25 30 Trp Met Asn Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Ser Asp Ser Glu Thr His Tyr Asn Gln Gln Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Ile Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ala 115 <210> 26 <211> 111 <212> PRT <213> Mouse (Mus sp.) <400> 26 Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Ile Ser Leu Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asp Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Thr Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Asp 85 90 95 Lys Asp Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110 <210> 27 <211> 117 <212> PRT <213> Mouse (Mus sp.) <400> 27 Gln Val Gln Leu Gln Gln Ser Gly Pro Gln Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Ala Leu Gln Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Leu Thr Val Ser Ser 115 <210> 28 <211> 107 <212> PRT <213> Mouse (Mus sp.) <400> 28 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Thr Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Ala Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Asn Arg Tyr Pro Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Met His Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Arg Pro Tyr Pro Phe 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 29 <211> 118 <212> PRT <213> Mouse (Mus sp.) <400> 29 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Val Arg Ser Gly Thr 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asn Pro Val Asn Gly Gly Ile Ser Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ile Arg Ser Gly Gly Asn Tyr Gly Gly Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 30 <211> 107 <212> PRT <213> Mouse (Mus sp.) <400> 30 Asp Val Val Val Thr Gln Thr Pro Lys Phe Leu Pro Val Thr Ala Glu 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Ser Asn Glu 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ser His Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Val 65 70 75 80 Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln His Phe Ser Ser Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 31 <211> 119 <212> PRT <213> Mouse (Mus sp.) <400> 31 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 20 25 30 Thr Met Asn Trp Val Lys Gln Ser Leu Gly Lys Asn Leu Glu Trp Ile 35 40 45 Gly Leu Ile Ser Pro Tyr Asn Gly Val Thr Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Ser Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Glu Leu Leu Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Val Thr Val Pro Leu Arg Phe Asp Val Trp Gly Ala Gly 100 105 110 Thr Pro Val Thr Val Ser Ser 115 <210> 32 <211> 113 <212> PRT <213> Mouse (Mus sp.) <400> 32 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Thr Gly 1 5 10 15 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Thr Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Asn Asn Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 85 90 95 Asp Asp Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys <210> 33 <211> 119 <212> PRT <213> Artificial Sequence <400> 33 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Asn 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ser Asp Ser Glu Thr His Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 34 <211> 111 <212> PRT <213> Artificial Sequence <400> 34 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Ile Ser Leu Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asp Leu Glu Ser Gly Val Pro Ser 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Ser 65 70 75 80 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Asp 85 90 95 Lys Asp Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 110 <210> 35 <211> 119 <212> PRT <213> Artificial Sequence <400> 35 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Asn 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ser Glu Ser Glu Thr His Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 36 <211> 117 <212> PRT <213> Artificial Sequence <400> 36 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Ala Leu Gln Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 37 <211> 107 <212> PRT <213> Artificial Sequence <400> 37 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Ala Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Asn Arg Tyr Pro Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Arg Pro Tyr Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 38 <211> 117 <212> PRT <213> Artificial Sequence <400> 38 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Glu Ser Glu Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Ala Leu Gln Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 39 <211> 118 <212> PRT <213> Artificial Sequence <400> 39 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asn Pro Val Asn Gly Gly Ile Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Asn Tyr Gly Gly Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 40 <211> 107 <212> PRT <213> Artificial Sequence <400> 40 Asp Val Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Asn Glu 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ser His Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His Phe Ser Ser Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 41 <211> 118 <212> PRT <213> Artificial Sequence <400> 41 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asn Pro Val Gln Gly Gly Ile Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Asn Tyr Gly Gly Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 42 <211> 119 <212> PRT <213> Artificial Sequence <400> 42 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 20 25 30 Thr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Ser Pro Tyr Asn Gly Val Thr Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Thr Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Val Thr Val Pro Leu Arg Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 43 <211> 113 <212> PRT <213> Artificial Sequence <400> 43 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys 35 40 45 Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Asn 85 90 95 Asp Asp Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 100 105 110 Lys <210> 44 <211> 119 <212> PRT <213> Artificial Sequence <400> 44 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 20 25 30 Thr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Ser Pro Tyr Gln Gly Val Thr Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Thr Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Val Thr Val Pro Leu Arg Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 45 <211> 113 <212> PRT <213> Artificial Sequence <400> 45 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Gln Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys 35 40 45 Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Asn 85 90 95 Asp Asp Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 100 105 110 Lys <210> 46 <211> 449 <212> PRT <213> Artificial Sequence <400> 46 Gln Val Gln Leu Gln Gln Pro Gly Val Glu Val Val Arg Pro Gly Ala 1 5 10 15 Pro Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Asn 20 25 30 Trp Met Asn Trp Val Lys Gln Arg Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Ser Asp Ser Glu Thr His Tyr Asn Gln Gln Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Ile Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 47 <211> 218 <212> PRT <213> Artificial Sequence <400> 47 Asp Ile Val Met Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Ile Ser Leu Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asp Leu Glu Ser Gly Ile Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Asn 65 70 75 80 Pro Val Glu Thr Asp Asp Val Ala Thr Tyr Tyr Cys Gln Gln Ser Asp 85 90 95 Lys Asp Pro Pro Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 145 150 155 160 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 48 <211> 1350 <212> DNA <213> Artificial Sequence <400> 48 caggtgcagc tgcagcagcc tggagtggag gtggtgagac caggagcccc tgtgaagctg 60 tcttgcaagg cttccggcta caccttcacc acaaactgga tgaattgggt gaagcagagg 120 ccaggaaggg gactggagtg gatcggacgc atcgacccca gcgattctga gacacactac 180 aaccagcagt ttaaggacaa ggccaccctg acagtggata agtccagcaa taccgcttat 240 atccagctgt cttccctgac atccgaggac agcgccgtgt actattgcgt gagggagggc 300 atctacggcc catggttcgc ctattgggga cagggcaccc tggtgacagt gtccgctgcc 360 tctacaaagg gcccttccgt gtttccactg gctccctcca gcaagtctac ctccggagga 420 acagccgctc tgggatgtct ggtgaaggac tatttcccag agcccgtgac cgtgtcttgg 480 aactccggcg ccctgacctc tggagtgcac acatttcctg ctgtgctgca gtcttccggc 540 ctgtactccc tgagctctgt ggtgaccgtg ccatccagct ctctgggcac ccagacatat 600 atctgcaacg tgaatcacaa gccatctaat acaaaggtgg acaagaaggt ggagcccaag 660 tcctgtgata agacccatac atgcccccct tgtcctgctc cagagctgct gggaggacca 720 tccgtgttcc tgtttccacc caagcctaag gacaccctga tgatcagcag aaccccagag 780 gtgacatgcg tggtggtggc agtgtctcac gaggatcccg aggtgaagtt taactggtac 840 gtggatggcg tggaggtgca taatgctaag acaaagccaa gggaggagca gtacaacagc 900 acctatcggg tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag 960 tataagtgca aggtgtccaa taaggccctg cccgctccta tcgagaagac catcagcaag 1020 gccaagggcc agcctagaga gccacaggtg tacacactgc ctccatcccg cgacgagctg 1080 accaagaacc aggtgagcct gacatgtctg gtgaagggct tctatcccag cgatatcgct 1140 gtggagtggg agtctaatgg ccagcctgag aacaattaca agaccacacc ccctgtgctg 1200 gacagcgatg gctctttctt tctgtattct aagctgaccg tggataagtc ccgctggcag 1260 cagggcaacg tgtttagctg ttctgtgatg catgaggctc tgcacaatca ttacacacag 1320 aagtccctga gcctgtctcc tggcaagtga 1350 <210> 49 <211> 657 <212> DNA <213> Artificial Sequence <400> 49 gacatcgtga tgacccagtc tccagcttcc ctggccgtgt ccctgggaca gagggctaca 60 atcagctgca gggcttccga gagcgtggat aactacggca tctctctgat gcactggtat 120 cagcagaagc ctggccagcc ccctaagctg ctgatctaca gggccagcga cctggagtct 180 ggcatccctg ctagattctc tggatccgga agcaggaccg acttcaccct gacaatcaat 240 ccagtggaga ccgacgatgt ggccacatac tattgtcagc agtccgacaa ggatccaccc 300 accttcggcg ctggcacaaa gctggagctg aagcgtacgg tggccgctcc ttccgtgttc 360 atctttcccc cttccgatga gcagctgaag agcggcacag cttctgtggt gtgcctgctg 420 aacaacttct acccaaggga ggccaaggtg cagtggaagg tggacaacgc tctgcagagc 480 ggcaattctc aggagtccgt gaccgagcag gacagcaagg attctacata ttccctgtcc 540 agcaccctga cactgtccaa ggccgattac gagaagcaca aggtgtatgc ttgcgaggtg 600 acccatcagg gcctgtcttc ccccgtgaca aagagcttta atagaggcga gtgttga 657 <210> 50 <211> 447 <212> PRT <213> Artificial Sequence <400> 50 Gln Val Gln Leu Gln Gln Ser Gly Pro Gln Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Asp Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Leu Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Ala Leu Gln Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr 100 105 110 Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 51 <211> 214 <212> PRT <213> Artificial Sequence <400> 51 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Thr Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Ala Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Asn Arg Tyr Pro Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Met His Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln Tyr Arg Pro Tyr Pro Phe 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 52 <211> 1344 <212> DNA <213> Artificial Sequence <400> 52 caggtgcagc tgcagcagtc cggaccacag ctggtgagac ctggagcctc tgtgaagatc 60 tcctgcaagg cttccggcta cagcttcacc tcttattgga tgcactgggt gaagcagagg 120 ccaggacagg gactggactg gatcggcatg atcgacccca gcgattctga gacacggctg 180 aaccagaagt ttaaggacaa ggccaccctg acagtggata agtccagctc taccgcttac 240 atgcagctgt ccagccccac atccgaggac agcgccgtgt actattgtgc ccgctgggct 300 ctgcagtact tcgattattg gggccagggc accacactga ccgtgtcttc cgcctctaca 360 aagggccctt ccgtgtttcc actggctccc tccagcaagt ctacctccgg aggaacagcc 420 gctctgggat gtctggtgaa ggactatttc ccagagcccg tgaccgtgtc ttggaactcc 480 ggcgccctga cctctggagt gcacattt cctgctgtgc tgcagctttc cggcctgtac 540 tcctgagct ctgtggtgac cgtgccatcc agctctctgg gcacccagac atatatctgc 600 aacgtgaatc acaagccatc tatacaaag gtggacaaga aggtggagcc caagtcctgt 660 gataagaccc atacatgccc cccttgtcct gctccagagc tgctgggagg accatccgtg 720 ttcctgtttc cacccaagcc taaggacacc ctgatgatca gcagaacccc agaggtgaca 780 tgcgtggtgg tggcagtgtc tcacgaggat cccgaggtga agtttaactg gtacgtggat 840 ggcgtggagg tgcataatgc taagacaaag ccaagggagg agcagtacaa cagcacctat 900 cgggtggtgt ctgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtataag 960 tgcaaggtgt ccaataaggc cctgcccgct cctatcgaga agaccatcag caaggccaag 1020 ggccagccta gagagccaca ggtgtacaca ctgcctccat cccgcgacga gctgaccaag 1080 aaccaggtga gcctgacatg tctggtgaag ggcttctatc ccagcgatat cgctgtggag 1140 tgggagtcta atggccagcc tgagaacaat tacaagacca caccccctgt gctggacagc 1200 gatggctctt tctttctgta ttctaagctg accgtggata agtcccgctg gcagcagggc 1260 aacgtgttta gctgttctgt gatgcatgag gctctgcaca atcattacac acagaagtcc 1320 ctgagcctgt ctcctggcaa gtga 1344 <210> 53 <211> 645 <212> DNA <213> Artificial Sequence <400> 53 gacatcgtga tgacccagtc ccagaagttc atgagcacca cagtgggcga tagagtgtct 60 atcacatgca aggcttccca gaacgtggga gctgctgtgg cttggtacca gcagaagcct 120 ggccagtccc caaagctgct gatctactcc gccagcaaca ggtatccagg agtgccagac 180 cggttcaccg gatctggatc cggcacagac ttcaccctga caatcagcaa tatgcactct 240 gaggacctgg ctgattattt ttgtcagcag taccgcccct atcctttcac ctttggcggc 300 ggcacaaagc tggagatcaa gcgtacggtg gccgctcctt ccgtgttcat ctttccccct 360 tccgatgagc agctgaagag cggcacagct tctgtggtgt gcctgctgaa caacttctac 420 ccaagggagg ccaaggtgca gtggaaggtg gacaacgctc tgcagagcgg caattctcag 480 gagtccgtga ccgagcagga cagcaaggat tctacatatt ccctgtccag caccctgaca 540 ctgtccaagg ccgattacga gaagcacaag gtgtatgctt gcgaggtgac ccatcagggc 600 ctgtcttccc ccgtgacaaa gagctttaat agaggcgagt gttga 645 <210> 54 <211> 448 <212> PRT <213> Artificial Sequence <400> 54 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Val Arg Ser Gly Thr 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asn Pro Val Asn Gly Gly Ile Ser Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ile Arg Ser Gly Gly Asn Tyr Gly Gly Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 55 <211> 214 <212> PRT <213> Artificial Sequence <400> 55 Asp Val Val Val Thr Gln Thr Pro Lys Phe Leu Pro Val Thr Ala Glu 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Ser Asn Glu 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ser His Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Val 65 70 75 80 Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln His Phe Ser Ser Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 56 <211> 1347 <212> DNA <213> Artificial Sequence <400> 56 caggtgcagc tgcagcagtc cggagctgag gtggtgagga gcggcacctc tgtgaaggtg 60 tcttgcaagg cctccggcta cgctttcaca aactatctga tcgagtgggt gaagcagagg 120 ccaggacagg gactggagtg gatcggcgtg atcaaccctg tgaatggcgg catcagctac 180 aatgagaagt tcaagggcaa ggccaccctg acagctgaca agtccagctc taccgtgtac 240 atgcagctgt ccagcctgac atccgacgat tccgccgtgt acttttgtat caggtctggc 300 ggcaactacg gcggcgatta ttggggccag ggcaccacac tgaccgtgtc ttccgcctct 360 acaaagggcc cttccgtgtt tccactggct ccctccagca agtctacctc cggaggaaca 420 gccgctctgg gatgtctggt gaaggactat ttcccagagc ccgtgaccgt gtcttggaac 480 tccggcgccc tgacctctgg agtgcacaca tttcctgctg tgctgcagtc ttccggcctg 540 tactccctga gctctgtggt gaccgtgcca tccagctctc tgggcaccca gacatatatc 600 tgcaacgtga atcacaagcc atctaataca aaggtggaca agaaggtgga gcccaagtcc 660 tgtgataaga cccatacatg ccccccttgt cctgctccag agctgctggg aggaccatcc 720 gtgttcctgt ttccacccaa gcctaaggac accctgatga tcagcagaac cccagaggtg 780 acatgcgtgg tggtggcagt gtctcacgag gatcccgagg tgaagtttaa ctggtacgtg 840 gatggcgtgg aggtgcataa tgctaagaca aagccaaggg aggagcagta caacagcacc 900 tatcgggtgg tgtctgtgct gacagtgctg caccaggact ggctgaacgg caaggagtat 960 aagtgcaagg tgtccaataa ggccctgccc gctcctatcg agaagaccat cagcaaggcc 1020 aagggccagc ctagagagcc acaggtgtac acactgcctc catcccgga cgagctgacc 1080 aagaaccagg tgagcctgac atgtctggtg aagggcttct atcccagcga tatcgctgtg 1140 gagtgggagt ctaatggcca gcctgagaac aattacaaga ccacaccccc tgtgctggac 1200 agcgatggct ctttctttct gtattctaag ctgaccgtgg ataagtcccg ctggcagcag 1260 ggcaacgtgt ttagctgttc tgtgatgcat gaggctctgc acaatcatta cacacagaag 1320 tccctgagcc tgtctcctgg caagtga 1347 <210> 57 <211> 645 <212> DNA <213> Artificial Sequence <400> 57 gacgtggtgg tgacccagac acccaagttc ctgcctgtga ccgccgagga tagggtgacc 60 atcacatgca aggccagcca gtccgtgtcc aacgaggtgg cttggtacca gcagaagcca 120 ggccagtccc ccaagctgct gatctactat gcttcccaca ggtatacagg agtgcctgac 180 cggttcaccg gaagcggatc tggcacagat ttcaccttta caatctccag cgtgcaggtg 240 gaggacctgg ccgtgtactt ctgtcagcag catttttctt ccccactgac ctttggcgct 300 ggcacaaagc tggagctgaa gcgtacggtg gccgctcctt ccgtgttcat ctttccccct 360 tccgatgagc agctgaagag cggcacagct tctgtggtgt gcctgctgaa caacttctac 420 ccaagggagg ccaaggtgca gtggaaggtg gacaacgctc tgcagagcgg caattctcag 480 gagtccgtga ccgagcagga cagcaaggat tctacatatt ccctgtccag caccctgaca 540 ctgtccaagg ccgattacga gaagcacaag gtgtatgctt gcgaggtgac ccatcagggc 600 ctgtcttccc ccgtgacaaa gagctttaat agaggcgagt gttga 645 <210> 58 <211> 449 <212> PRT <213> Artificial Sequence <400> 58 Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Met Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 20 25 30 Thr Met Asn Trp Val Lys Gln Ser Leu Gly Lys Asn Leu Glu Trp Ile 35 40 45 Gly Leu Ile Ser Pro Tyr Asn Gly Val Thr Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Ser Leu Thr Val Asp Thr Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Glu Leu Leu Ser Leu Thr Ser Asp Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Val Thr Val Pro Leu Arg Phe Asp Val Trp Gly Ala Gly 100 105 110 Thr Pro Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 59 <211> 220 <212> PRT <213> Artificial Sequence <400> 59 Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Thr Gly 1 5 10 15 Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Thr Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Asn Asn Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 85 90 95 Asp Asp Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100 105 110 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 115 120 125 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 180 185 190 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 60 <211> 1350 <212> DNA <213> Artificial Sequence <400> 60 gaggtgcagc tgcagcagtc tggaccagag ctggtgaagc ctggagcttc catgaagatc 60 agctgcaagg cttctggcta cagcttctct ggctatacca tgaactgggt gaagcagtcc 120 ctgggcaaga atctggagtg gatcggcctg atcagcccct acaacggcgt gaccacatat 180 aatcagaagt ttaagggcaa ggcctccctg accgtggaca catccagcaa caccgcttac 240 atggagctgc tgtccctgac aagcgacgat tctgccgtgt actattgtgc taggggcgtg 300 acagtgccac tgcggttcga cgtgtggggc gccggcaccc ccgtgacagt gtcttccgcc 360 tctacaaagg gcccttccgt gtttccactg gctccctcca gcaagtctac ctccggagga 420 acagccgctc tgggatgtct ggtgaaggac tatttcccag agcccgtgac cgtgtcttgg 480 aactccggcg ccctgacctc tggagtgcac acatttcctg ctgtgctgca gtcttccggc 540 ctgtactccc tgagctctgt ggtgaccgtg ccatccagct ctctgggcac ccagacatat 600 atctgcaacg tgaatcacaa gccatctaat acaaaggtgg acaagaaggt ggagcccaag 660 tcctgtgata agacccatac atgcccccct tgtcctgctc cagagctgct gggaggacca 720 tccgtgttcc tgtttccacc caagcctaag gacaccctga tgatcagcag aaccccagag 780 gtgacatgcg tggtggtggc agtgtctcac gaggatcccg aggtgaagtt taactggtac 840 gtggatggcg tggaggtgca taatgctaag acaaagccaa gggaggagca gtacaacagc 900 acctatcggg tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag 960 tataagtgca aggtgtccaa taaggccctg cccgctccta tcgagaagac catcagcaag 1020 gccaagggcc agcctagaga gccacaggtg tacacactgc ctccatcccg cgacgagctg 1080 accaagaacc aggtgagcct gacatgtctg gtgaagggct tctatcccag cgatatcgct 1140 gtggagtggg agtctaatgg ccagcctgag aacaattaca agaccacacc ccctgtgctg 1200 gacagcgatg gctctttctt tctgtattct aagctgaccg tggataagtc ccgctggcag 1260 cagggcaacg tgtttagctg ttctgtgatg catgaggctc tgcacaatca ttacacacag 1320 aagtccctga gcctgtctcc tggcaagtga 1350 <210> 61 <211> 663 <212> DNA <213> Artificial Sequence <400> 61 gacatcgtga tgacccagag cccctccagc ctgacagtga ccacaggcga gaaggtgacc 60 atgtcttgca agtcttccca gtccctgctg aacagcggca atcagaagaa ctacctgacc tggtatcagc agaagacagg ccagccccct aagctgctga tctactgggc ctctacaagg gagtccggag tgccagaccg gttcaccgga agcggatctg gcacagactt caccctgaca 240 atcaacaatg tgcaggccga ggatctggcc gtgtactatt gtcagaatga cgattcctat cccctgacct tcggcgctgg cacaaagctg gagctgaagc gtacggtggc cgctccttcc 360 gtgttcatct ttcccccttc cgatgagcag ctgaagagcg gcacagcttc tgtggtgtgc 420 ctgctgaaca acttctaccc aagggaggcc aaggtgcagt ggaaggtgga caacgctctg cagagcggca attctcagga gtccgtgacc gagcaggaca gcaaggattc tacatattcc ctgtccagca ccctgacact gtccaaggcc gattacgaga agcacaaggt gtatgcttgc gaggtgaccc atcagggcct gtcttccccc gtgacaaaga gctttaatag aggcgagtgt 660 tga 663 <210> 62 <211> 449 <212> PRT <213> Artificial Sequence <400> 62 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Asn 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ser Asp Ser Glu Thr His Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 63 <211> 218 <212> PRT <213> Artificial Sequence <400> 63 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Asp Asn Tyr 20 25 30 Gly Ile Ser Leu Met His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 35 40 45 Lys Leu Leu Ile Tyr Arg Ala Ser Asp Leu Glu Ser Gly Val Pro Ser 50 55 60 Arg Phe Ser Gly Ser Gly Ser Arg Thr Asp Phe Thr Leu Thr Ile Ser 65 70 75 80 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Asp 85 90 95 Lys Asp Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 110 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115 120 125 Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 130 135 140 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 145 150 155 160 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 165 170 175 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180 185 190 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 195 200 205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 64 <211> 449 <212> PRT <213> Artificial Sequence <400> 64 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Asn 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Arg Ile Asp Pro Ser Glu Ser Glu Thr His Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Val Arg Glu Gly Ile Tyr Gly Pro Trp Phe Ala Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 65 <211> 1350 <212> DNA <213> Artificial Sequence <400> 65 caggtgcagc tggtgcagtc tggcgccgag gtgaagaagc caggcgcttc cgtgaaggtg 60 agctgcaagg cctctggcta caccttcacc acaaactgga tgaattgggt gagacaggct 120 ccaggacagg gactggagtg gatgggaagg atcgacccta gcgattctga gacacactac 180 gcccagaagt ttcagggcag ggtgaccatg acagtggaca agtccaccaa cacagtgtat 240 atggagctgt ccagcctgag gagcgaggat accgccgtgt actattgcgt gagggagggc 300 atctacggcc cttggttcgc ttattggggc cagggcaccc tggtgacagt gtcttccgcc 360 tctacaaagg gcccttccgt gtttccactg gctccctcca gcaagtctac ctccggagga 420 acagccgctc tgggatgtct ggtgaaggac tatttcccag agcccgtgac cgtgtcttgg 480 aactccggcg ccctgacctc tggagtgcac acatttcctg ctgtgctgca gtcttccggc 540 ctgtactccc tgagctctgt ggtgaccgtg ccatccagct ctctgggcac ccagacatat 600 atctgcaacg tgaatcacaa gccatctaat acaaaggtgg acaagaaggt ggagcccaag 660 tcctgtgata agacccatac atgcccccct tgtcctgctc cagagctgct gggaggacca 720 tccgtgttcc tgtttccacc caagcctaag gacaccctga tgatcagcag aaccccagag 780 gtgacatgcg tggtggtggc agtgtctcac gaggatcccg aggtgaagtt taactggtac 840 gtggatggcg tggaggtgca taatgctaag acaaagccaa gggaggagca gtacaacagc 900 acctatcggg tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag 960 tataagtgca aggtgtccaa taaggccctg cccgctccta tcgagaagac catcagcaag 1020 gccaagggcc agcctagaga gccacaggtg tacacactgc ctccatcccg cgacgagctg 1080 accaagaacc aggtgagcct gacatgtctg gtgaagggct tctatcccag cgatatcgct 1140 gtggagtggg agtctaatgg ccagcctgag aacaattaca agaccacacc ccctgtgctg 1200 gacagcgatg gctctttctt tctgtattct aagctgaccg tggataagtc ccgctggcag 1260 cagggcaacg tgtttagctg ttctgtgatg catgaggctc tgcacaatca ttacacacag 1320 aagtccctga gcctgtctcc tggcaagtga 1350 <210> 66 <211> 1350 <212> DNA <213> Artificial Sequence <400> 66 caggtgcagc tggtgcagtc tggcgccgag gtgaagaagc caggcgcttc cgtgaaggtg 60 agctgcaagg cctctggcta caccttcacc acaaactgga tgaattgggt gagacaggct 120 ccaggacagg gactggagatg gatgggaagg atcgacccta gcgagtctga gacacactac 180 gcccagaagt ttcagggcag ggtgaccatg acagtggaca agtccaccaa cacagtgtat 240 atggagctgt ccagcctgag gagcgaggat accgccgtgt actattgcgt gagggagggc 300 atctacggcc cttggttcgc ttatggggc cagggcaccc tggtgacagt gtttccgcc 360 tctacaaagg gcccttccgt gtttccactg gctccctcca gcaagtctac ctccggagga 420 acagccgctc tgggatgtct ggtgaaggac tatttcccag agcccgtgac cgtgtcttgg 480 aactccggcg ccctgacctc tggagtgcac acatttcctg ctgtgctgca gtcttccggc 540 ctgtactccc tgagctctgt ggtgaccgtg ccatccagct ctctgggcac ccagacatat 600 atctgcaacg tgaatcacaa gccatctaat acaaaggtgg acaagaaggt ggagcccaag 660 tcctgtgata agacccatac atgcccccct tgtcctgctc cagagctgct gggaggacca 720 tccgtgttcc tgtttccacc caagcctaag gacaccctga tgatcagcag aaccccagag 780 gtgacatgcg tggtggtggc agtgtctcac gaggatcccg aggtgaagtt taactggtac 840 gtggatggcg tggaggtgca taatgctaag acaaagccaa gggaggagca gtacaacagc 900 acctatcggg tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag 960 tataagtgca aggtgtccaa taaggccctg cccgctccta tcgagaagac catcagcaag 1020 gccaagggcc agcctagaga gccacaggtg tacacactgc ctccatcccg cgacgagctg 1080 accaagaacc aggtgagcct gacatgtctg gtgaagggct tctatcccag cgatatcgct 1140 gtggagtggg agtctaatgg ccagcctgag aacaattaca agaccacacc ccctgtgctg 1200 gacagcgatg gctctttctt tctgtattct aagctgaccg tggataagtc ccgctggcag 1260 cagggcaacg tgtttagctg ttctgtgatg catgaggctc tgcacaatca ttacacacag 1320 aagtccctga gcctgtctcc tggcaagtga 1350 <210> 67 <211> 657 <212> DNA <213> Artificial Sequence <400> 67 gacatccaga tgacccagtc tccatccagc ctgagcgcct ctgtgggcga cagggtgacc 60 atcacatgcc gggcttccga gagcgtggat aactacggca tctccctgat gcactggtat 120 cagcagaagc caggcaaggc ccccaagctg ctgatctaca gggcttctga tctggagtcc 180 ggcgtgccta gcagattctc tggctccggc agccgcaccg actttaccct gacaatctct 240 tccctgcagc cagaggattt cgccacatac tattgtcagc agtccgacaa ggatccccct 300 acctttggcc agggcacaaa ggtggagatc aagcgtacgg tggccgctcc ttccgtgttc 360 atctttcccc cttccgatga gcagctgaag agcggcacag cttctgtggt gtgcctgctg 420 aacaacttct acccaaggga ggccaaggtg cagtggaagg tggacaacgc tctgcagagc 480 ggcaattctc aggagtccgt gaccgagcag gacagcaagg attctacata ttccctgtcc 540 agcaccctga cactgtccaa ggccgattac gagaagcaca aggtgtatgc ttgcgaggtg 600 acccatcagg gcctgtcttc ccccgtgaca aagagcttta atagaggcga gtgttga 657 <210> 68 <211> 447 <212> PRT <213> Artificial Sequence <400> 68 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Ala Leu Gln Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 69 <211> 214 <212> PRT <213> Artificial Sequence <400> 69 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Ala Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Asn Arg Tyr Pro Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Arg Pro Tyr Pro Phe 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 70 <211> 447 <212> PRT <213> Artificial Sequence <400> 70 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Met Ile Asp Pro Ser Glu Ser Glu Thr Arg Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Ala Leu Gln Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 71 <211> 1344 <212> DNA <213> Artificial Sequence <400> 71 caggtgcagc tggtgcagag cggcgctgag gtgaagaagc caggagcttc tgtgaaggtg 60 tcctgcaagg cctctggcta ctccttcacc agctattgga tgcactgggt gaggcaggct 120 ccaggacagg gactggagtg gatgggaatg atcgacccta gcgattctga gacaagatac 180 gcccagaagt ttcagggccg ggtgaccatg acagtggaca agtccaccag cacagtgtat 240 atggagctgt ccagcctgag atctgaggac accgccgtgt actattgtgc ccgctgggct 300 ctgcagtact tcgattattg gggccagggc accctggtga cagtgtcttc cgcctctaca 360 aagggccctt ccgtgtttcc actggctccc tccagcaagt ctacctccgg aggaacagcc 420 gctctgggat gtctggtgaa ggactatttc ccagagcccg tgaccgtgtc ttggaactcc 480 ggcgccctga cctctggagt gcacattt cctgctgtgc tgcagctttc cggcctgtac 540 tcctgagct ctgtggtgac cgtgccatcc agctctctgg gcacccagac atatatctgc 600 aacgtgaatc acaagccatc tatacaaag gtggacaaga aggtggagcc caagtcctgt 660 gataagaccc atacatgccc cccttgtcct gctccagagc tgctgggagg accatccgtg 720 ttcctgtttc cacccaagcc taaggacacc ctgatgatca gcagaacccc agaggtgaca 780 tgcgtggtgg tggcagtgtc tcacgaggat cccgaggtga agtttaactg gtacgtggat 840 ggcgtggagg tgcataatgc taagacaaag ccaagggagg agcagtacaa cagcacctat 900 cgggtggtgt ctgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtataag 960 tgcaaggtgt ccaataaggc cctgcccgct cctatcgaga agaccatcag caaggccaag 1020 ggccagccta gagagccaca ggtgtacaca ctgcctccat cccgcgacga gctgaccaag 1080 aaccaggtga gcctgacatg tctggtgaag ggcttctatc ccagcgatat cgctgtggag 1140 tgggagtcta atggccagcc tgagaacaat tacaagacca caccccctgt gctggacagc 1200 gatggctctt tctttctgta ttctaagctg accgtggata agtcccgctg gcagcagggc 1260 aacgtgttta gctgttctgt gatgcatgag gctctgcaca atcattacac acagaagtcc 1320 ctgagcctgt ctcctggcaa gtga 1344 <210> 72 <211> 1344 <212> DNA <213> Artificial Sequence <400> 72 caggtgcagc tggtgcagag cggcgctgag gtgaagaagc caggagcttc tgtgaaggtg 60 tcctgcaagg cctctggcta ctccttcacc agctattgga tgcactgggt gaggcaggct 120 ccaggacagg gactggagtg gatgggaatg atcgacccta gcgagtctga gacaagatac 180 gcccagaagt ttcagggccg ggtgaccatg acagtggata agtccaccag cacagtgtat 240 atggagctgt ccagcctgag atctgaggac accgccgtgt actattgtgc ccgctgggct 300 ctgcagtact tcgattattg gggccagggc accctggtga cagtgtcttc cgcctctaca 360 aagggccctt ccgtgtttcc actggctccc tccagcaagt ctacctccgg aggaacagcc 420 gctctgggat gtctggtgaa ggactatttc ccagagcccg tgaccgtgtc ttggaactcc 480 ggcgccctga cctctggagt gcacattt cctgctgtgc tgcagctttc cggcctgtac 540 tcctgagct ctgtggtgac cgtgccatcc agctctctgg gcacccagac atatatctgc 600 aacgtgaatc acaagccatc tatacaaag gtggacaaga aggtggagcc caagtcctgt 660 gataagaccc atacatgccc cccttgtcct gctccagagc tgctgggagg accatccgtg 720 ttcctgtttc cacccaagcc taaggacacc ctgatgatca gcagaacccc agaggtgaca 780 tgcgtggtgg tggcagtgtc tcacgaggat cccgaggtga agtttaactg gtacgtggat 840 ggcgtggagg tgcataatgc taagacaaag ccaagggagg agcagtacaa cagcacctat 900 cgggtggtgt ctgtgctgac agtgctgcac caggactggc tgaacggcaa ggagtataag 960 tgcaaggtgt ccaataaggc cctgcccgct cctatcgaga agaccatcag caaggccaag 1020 ggccagccta gagagccaca ggtgtacaca ctgcctccat cccgcgacga gctgaccaag 1080 aaccaggtga gcctgacatg tctggtgaag ggcttctatc ccagcgatat cgctgtggag 1140 tgggagtcta atggccagcc tgagaacaat tacaagacca caccccctgt gctggacagc 1200 gatggctctt tctttctgta ttctaagctg accgtggata agtcccgctg gcagcagggc 1260 aacgtgttta gctgttctgt gatgcatgag gctctgcaca atcattacac acagaagtcc 1320 ctgagcctgt ctcctggcaa gtga 1344 <210> 73 <211> 645 <212> DNA <213> Artificial Sequence <400> 73 gacatccaga tgacccagtc tccctccaca ctgagcgcct ctgtgggcga tagagtgacc 60 gacatccaga tgacccagtc tccctccaca ctgagcgcct ctgtgggcga tagagtgacc 60 atcacatgca aggcctctca gaacgtggga gctgctgtgg cttggtacca gcagaagcct 120 atcacatgca aggcctctca gaacgtggga gctgctgtgg cttggtacca gcagaagcct 120 ggcaaggccc caaagctgct gatctactcc gctagcaata ggtatccagg agtgccatcc 180 ggcaaggccc caaagctgct gatctactcc gctagcaata ggtatccagg agtgccatcc 180 cggttctctg gctccggcag cggcaccgag tttaccctga caatctccag cctgcagcct 240 cggttctctg gctccggcag cggcaccgag tttaccctga caatctccag cctgcagcct 240 gacgatttcg ctacatacta ttgtcagcag taccgcccct atcctttcac ctttggccag 300 gacgatttcg ctacatacta ttgtcagcag taccgcccct atcctttcac ctttggccag 300 ggcacaaagg tggagatcaa gcgtacggtg gccgctcctt ccgtgttcat ctttccccct 360 ggcacaaagg tggagatcaa gcgtacggtg gccgctcctt ccgtgttcat ctttccccct 360 tccgatgagc agctgaagag cggcacagct tctgtggtgt gcctgctgaa caacttctac 420 tccgatgagc agctgaagag cggcacagct tctgtggtgt gcctgctgaa caacttctac 420 ccaagggagg ccaaggtgca gtggaaggtg gacaacgctc tgcagagcgg caattctcag 480 ccaagggagg ccaaggtgca gtggaaggtg gacaacgctc tgcagagcgg caattctcag 480 gagtccgtga ccgagcagga cagcaaggat tctacatatt ccctgtccag caccctgaca 540 gagtccgtga ccgagcagga cagcaaggat tctacatatt ccctgtccag caccctgaca 540 ctgtccaagg ccgattacga gaagcacaag gtgtatgctt gcgaggtgac ccatcagggc 600 ctgtccaagg ccgattacga gaagcacaag gtgtatgctt gcgaggtgac ccatcagggc 600 ctgtcttccc ccgtgacaaa gagctttaat agaggcgagt gttga 645 ctgtcttccc ccgtgacaaa gagctttaat agaggcgagt gttga 645 <210> 74 <210> 74 <211> 448 <211> 448 <212> PRT <212> PRT <213> 人工序列(Artificial Sequence) <213> Artificial Sequence <400> 74 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asn Pro Val Asn Gly Gly Ile Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Asn Tyr Gly Gly Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 75 <211> 214 <212> PRT <213> Artificial Sequence <400> 75 Asp Val Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Ser Cys Lys Ala Ser Gln Ser Val Ser Asn Glu 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ser His Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln His Phe Ser Ser Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 76 <211> 448 <212> PRT <213> Artificial Sequence <400> 76 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Thr Asn Tyr 20 25 30 Leu Ile Glu Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Val Ile Asn Pro Val Gln Gly Gly Ile Ser Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Gly Gly Asn Tyr Gly Gly Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 77 <211> 1347 <212> DNA <213> Artificial Sequence <400> 77 caggtgcagc tggtgcagtc tggcgccgag gtgaagaagc caggcgcttc cgtgaaggtg 60 agctgcaagg cctctggcta cgctttcacc aactatctga tcgagtgggt gagacaggct 120 ccaggacagg gactggagtg gatgggcgtg atcaaccctg tgaatggcgg catctcctac 180 gcccagaagt ttcagggccg cgtgaccatg acagctgaca agagcacctc tacagtgtat 240 atggagctgt ccagcctgag gtccgaggac acagccgtgt actattgtgc tcggagcggc 300 ggaaattacg gaggcgatta ttggggccag ggcaccctgg tgacagtgtc ttccgcctct 360 acaaagggcc cttccgtgtt tccactggct ccctccagca agtctacctc cggaggaaca 420 gccgctctgg gatgtctggt gaaggactat ttcccagagc ccgtgaccgt gtcttggaac 480 tccggcgccc tgacctctgg agtgcacaca tttcctgctg tgctgcagtc ttccggcctg 540 tactccctga gctctgtggt gaccgtgcca tccagctctc tgggcaccca gacatatatc 600 tgcaacgtga atcacaagcc atctaataca aaggtggaca agaaggtgga gcccaagtcc 660 tgtgataaga cccatacatg ccccccttgt cctgctccag agctgctggg aggaccatcc 720 gtgttcctgt ttccacccaa gcctaaggac accctgatga tcagcagaac cccagaggtg 780 acatgcgtgg tggtggcagt gtctcacgag gatcccgagg tgaagtttaa ctggtacgtg 840 gatggcgtgg aggtgcataa tgctaagaca aagccaaggg aggagcagta caacagcacc 900 tatcgggtgg tgtctgtgct gacagtgctg caccaggact ggctgaacgg caaggagtat 960 aagtgcaagg tgtccaataa ggccctgccc gctcctatcg agaagaccat cagcaaggcc 1020 aagggccagc ctagagagcc acaggtgtac acactgcctc catcccgcga cgagctgacc 1080 aagaaccagg tgagcctgac atgtctggtg aagggcttct atcccagcga tatcgctgtg 1140 gagtgggagt ctaatggcca gcctgagaac aattacaaga ccacaccccc tgtgctggac 1200 agcgatggct ctttctttct gtattctaag ctgaccgtgg ataagtcccg ctggcagcag 1260 ggcaacgtgt ttagctgttc tgtgatgcat gaggctctgc acaatcatta cacacagaag 1320 tccctgagcc tgtctcctgg caagtga 1347 <210> 78 <211> 1347 <212> DNA <213> Artificial Sequence <400> 78 caggtgcagc tggtgcagtc tggcgccgag gtgaagaagc caggcgcttc cgtgaaggtg 60 agctgcaagg cctctggcta cgctttcacc aactatctga tcgagtgggt gagacaggct 120 ccaggacagg gactggagtg gatgggcgtg atcaatcctg tgcagggcgg catctcctac 180 gcccagaagt ttcagggccg cgtgaccatg acagctgaca agagcacctc tacagtgtat 240 atggagctgt ccagcctgag gtccgaggac acagccgtgt actattgtgc tcggagcggc 300 ggaaactacg gaggcgatta ttggggccag ggcaccctgg tgacagtgtc ttccgcctct 360 acaaagggcc cttccgtgtt tccactggct ccctccagca agtctacctc cggaggaaca 420 gccgctctgg gatgtctggt gaaggactat ttcccagagc ccgtgaccgt gtcttggaac 480 tccggcgccc tgacctctgg agtgcacaca tttcctgctg tgctgcagtc ttccggcctg 540 tactccctga gctctgtggt gaccgtgcca tccagctctc tgggcaccca gacatatatc 600 tgcaacgtga atcacaagcc atctaataca aaggtggaca agaaggtgga gcccaagtcc 660 tgtgataaga cccatacatg ccccccttgt cctgctccag agctgctggg aggaccatcc 720 gtgttcctgt ttccacccaa gcctaaggac accctgatga tcagcagaac cccagaggtg 780 acatgcgtgg tggtggcagt gtctcacgag gatcccgagg tgaagtttaa ctggtacgtg 840 gatggcgtgg aggtgcataa tgctaagaca aagccaaggg aggagcagta caacagcacc 900 tatcgggtgg tgtctgtgct gacagtgctg caccaggact ggctgaacgg caaggagtat 960 aagtgcaagg tgtccaataa ggccctgccc gctcctatcg agaagaccat cagcaaggcc 1020 aagggccagc ctagagagcc acaggtgtac acactgcctc catcccgcga cgagctgacc 1080 aagaaccagg tgagcctgac atgtctggtg aagggcttct atcccagcga tatcgctgtg 1140 gagtgggagt ctaatggcca gcctgagaac aattacaaga ccacaccccc tgtgctggac 1200 agcgatggct ctttctttct gtattctaag ctgaccgtgg ataagtcccg ctggcagcag 1260 ggcaacgtgt ttagctgttc tgtgatgcat gaggctctgc acaatcatta cacacagaag 1320 tccctgagcc tgtctcctgg caagtga 1347 <210> 79 <211> 645 <212> DNA <213> Artificial Sequence <400> 79 gacgtggtca tgacccagag ccctgattct ctggccgtgt ccctgggaga gagggctaca 60 atcagctgca aggctagcca gtccgtgtcc aacgaggtgg cctggtacca gcagaagcca 120 ggccagcccc ctaagctgct gatctactat gcttctcaca ggtataccgg cgtgcccgac 180 aggttctccg gctctggctc cggcacagac ttcaccctga caatctccag cctgcaggct 240 gaggacgtgg ccgtgtacta ttgtcagcag catttctctt ccccactgac ctttggccag 300 ggcacaaagg tggagatcaa gcgtacggtg gccgctcctt ccgtgttcat ctttccccct 360 tccgatgagc agctgaagag cggcacagct tctgtggtgt gcctgctgaa caacttctac 420 ccaagggagg ccaaggtgca gtggaaggtg gacaacgctc tgcagagcgg caattctcag 480 gagtccgtga ccgagcagga cagcaaggat tctacatatt ccctgtccag caccctgaca 540 ctgtccaagg ccgattacga gaagcacaag gtgtatgctt gcgaggtgac ccatcagggc 600 ctgtcttccc ccgtgacaaa gagctttaat agaggcgagt gttga 645 <210> 80 <211> 449 <212> PRT <213> Artificial Sequence <400> 80 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 20 25 30 Thr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Ser Pro Tyr Asn Gly Val Thr Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Thr Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Val Thr Val Pro Leu Arg Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 81 <211> 220 <212> PRT <213> Artificial Sequence <400> 81 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys 35 40 45 Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Asn 85 90 95 Asp Asp Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 100 105 110 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 115 120 125 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 180 185 190 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 82 <211> 449 <212> PRT <213> Artificial Sequence <400> 82 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 20 25 30 Thr Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Leu Ile Ser Pro Tyr Gln Gly Val Thr Thr Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Met Thr Val Asp Thr Ser Thr Asn Thr Val Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Val Thr Val Pro Leu Arg Phe Asp Val Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 83 <211> 220 <212> PRT <213> Artificial Sequence <400> 83 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Gln Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys 35 40 45 Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Asn 85 90 95 Asp Asp Ser Tyr Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 100 105 110 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 115 120 125 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 180 185 190 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 84 <211> 1350 <212> DNA <213> Artificial Sequence <400> 84 caggtgcagc tggtgcagtc tggcgccgag gtgaagaagc caggcgcttc cgtgaaggtg 60 agctgcaagg cctctggcta cagcttctct ggctatacaa tgaactgggt gaggcaggct 120 ccaggacagg gactggagtg gatgggcctg atctcccctt acaatggcgt gaccacatat 180 gcccagaagt ttcagggcag agtgaccatg acagtggaca cctccacaaa caccgtgtac 240 atggagctgt ccagcctgcg cagcgaggat acagccgtgt actattgtgc taggggagtg 300 accgtgccac tgcggttcga cgtgtgggga cagggcacac tggtgaccgt gtcttccgcc 360 tctacaaagg gcccttccgt gtttccactg gctccctcca gcaagtctac ctccggagga 420 acagccgctc tgggatgtct ggtgaaggac tatttcccag agcccgtgac cgtgtcttgg 480 aactccggcg ccctgacctc tggagtgcac acatttcctg ctgtgctgca gtcttccggc 540 ctgtactccc tgagctctgt ggtgaccgtg ccatccagct ctctgggcac ccagacatat 600 atctgcaacg tgaatcacaa gccatctaat acaaaggtgg acaagaaggt ggagcccaag 660 tcctgtgata agacccatac atgcccccct tgtcctgctc cagagctgct gggaggacca 720 tccgtgttcc tgtttccacc caagcctaag gacaccctga tgatcagcag aaccccagag 780 gtgacatgcg tggtggtggc agtgtctcac gaggatcccg aggtgaagtt taactggtac 840 gtggatggcg tggaggtgca taatgctaag acaaagccaa gggaggagca gtacaacagc 900 acctatcggg tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag 960 tataagtgca aggtgtccaa taaggccctg cccgctccta tcgagaagac catcagcaag 1020 gccaagggcc agcctagaga gccacaggtg tacacactgc ctccatcccg cgacgagctg 1080 accaagaacc aggtgagcct gacatgtctg gtgaagggct tctatcccag cgatatcgct 1140 gtggagtggg agtctaatgg ccagcctgag aacaattaca agaccacacc ccctgtgctg 1200 gacagcgatg gctctttctt tctgtattct aagctgaccg tggataagtc ccgctggcag 1260 cagggcaacg tgtttagctg ttctgtgatg catgaggctc tgcacaatca ttacacacag 1320 aagtccctga gcctgtctcc tggcaagtga 1350 <210> 85 <211> 1350 <212> DNA <213> Artificial Sequence <400> 85 caggtgcagc tggtgcagtc tggcgccgag gtgaagaagc caggcgcttc cgtgaaggtg 60 agctgcaagg cctctggcta cagcttctct ggctatacaa tgaactgggt gaggcaggct 120 ccaggacagg gactggagtg gatgggcctg atctcccctt accagggcgt gaccacatat 180 gcccagaagt ttcagggcag agtgaccatg acagtggaca cctccacaaa taccgtgtac 240 atggagctgt ccagcctgcg cagcgaggat acagccgtgt actattgtgc taggggagtg 300 accgtgccac tgcggttcga cgtgtgggga cagggcacac tggtgaccgt gtcttccgcc 360 tctacaaagg gcccttccgt gtttccactg gctccctcca gcaagtctac ctccggagga 420 acagccgctc tgggatgtct ggtgaaggac tatttcccag agcccgtgac cgtgtcttgg 480 aactccggcg ccctgacctc tggagtgcac acatttcctg ctgtgctgca gtcttccggc 540 ctgtactccc tgagctctgt ggtgaccgtg ccatccagct ctctgggcac ccagacatat 600 atctgcaacg tgaatcacaa gccatctaat acaaaggtgg acaagaaggt ggagcccaag 660 tcctgtgata agacccatac atgcccccct tgtcctgctc cagagctgct gggaggacca 720 tccgtgttcc tgtttccacc caagcctaag gacaccctga tgatcagcag aaccccagag 780 gtgacatgcg tggtggtggc agtgtctcac gaggatcccg aggtgaagtt taactggtac 840 gtggatggcg tggaggtgca taatgctaag acaaagccaa gggaggagca gtacaacagc 900 acctatcggg tggtgtctgt gctgacagtg ctgcaccagg actggctgaa cggcaaggag 960 tataagtgca aggtgtccaa taaggccctg cccgctccta tcgagaagac catcagcaag 1020 gccaagggcc agcctagaga gccacaggtg tacacactgc ctccatcccg cgacgagctg 1080 accaagaacc aggtgagcct gacatgtctg gtgaagggct tctatcccag cgatatcgct 1140 gtggagtggg agtctaatgg ccagcctgag aacaattaca agaccacacc ccctgtgctg 1200 gacagcgatg gctctttctt tctgtattct aagctgaccg tggataagtc ccgctggcag 1260 cagggcaacg tgtttagctg ttctgtgatg catgaggctc tgcacaatca ttacacacag 1320 aagtccctga gcctgtctcc tggcaagtga 1350 <210> 86 <211> 663 <212> DNA <213> Artificial Sequence <400> 86 gacatccaga tgacacagag cccttccagc ctgtccgcca gcgtgggcga tagggtgacc 60 atcacatgca agtcttccca gtctctgctg aactccggca atcagaagaa ctacctgacc 120 tggtatcagc agaagcccgg caaggcccct aagctgctga tctactgggc tagcaccagg 180 gagtctggcg tgccatcccg gttctctggc tccggcagcg gcacagactt taccctgaca 240 atcagctctc tgcagccaga ggatttcgct acctactatt gtcagaatga cgattcttat 300 cccctgacct ttggccaggg cacaaaggtg gagatcaagc gtacggtggc cgctccttcc 360 gtgttcatct ttcccccttc cgatgagcag ctgaagagcg gcacagcttc tgtggtgtgc 420 ctgctgaaca acttctaccc aagggaggcc aaggtgcagt ggaaggtgga caacgctctg cagagcggca attctcagga gtccgtgacc gagcaggaca gcaaggattc tacatattcc ctgtccagca ccctgacact gtccaaggcc gattacgaga agcacaaggt gtatgcttgc gaggtgaccc atcagggcct gtcttccccc gtgacaaaga gctttaatag aggcgagtgt 660 tga 663 <210> 87 <211> 663 <212> DNA <213> Artificial Sequence <400> 87 60. gacatccaga tgacacagag cccttccagc ctgtccgcca gcgtgggcga tagggtgacc atcacatgca agtcttccca gtctctgctg cagtccggca accagaagaa ttacctgacc tggtatcagc agaagcccgg caaggcccct aagctgctga tctactgggc tagcaccagg 180 gagtctggcg tgccatcccg gttctctggc tccggcagcg gcacagactt taccctgaca 240 atcagctctc tgcagccaga ggatttcgct acctactatt gtcagaacga cgattcttat cccctgacct ttggccaggg cacaaaggtg gagatcaagc gtacggtggc cgctccttcc 360 gtgttcatct ttcccccttc cgatgagcag ctgaagagcg gcacagcttc tgtggtgtgc 420 ctgctgaaca acttctaccc aagggaggcc aaggtgcagt ggaaggtgga caacgctctg 480 cagagcggca attctcagga gtccgtgacc gagcaggaca gcaaggattc tacatattcc 540 ctgtccagca ccctgacact gtccaaggcc gattacgaga agcacaaggt gtatgcttgc 600 gaggtgaccc atcagggcct gtcttccccc gtgacaaaga gctttaatag aggcgagtgt 660 tga 663 <210> 88 <211> 446 <212> PRT <213> Artificial Sequence <400> 88 Gln Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Thr Tyr 20 25 30 Gly Val His Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu 35 40 45 Gly Val Ile Trp Arg Gly Val Thr Thr Asp Tyr Asn Ala Ala Phe Met 50 55 60 Ser Arg Leu Thr Ile Thr Lys Asp Asn Ser Lys Asn Gln Val Val Leu 65 70 75 80 Thr Met Asn Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala 85 90 95 Arg Leu Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 89 <211> 214 <212> PRT <213> Artificial Sequence <400> 89 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Ser Asn Asp 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Ala Ala Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Asp Tyr Thr Ser Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210

Claims

1. An isolated anti-PD-L2 antibody or antigen-binding fragment thereof, comprising: The heavy chain CDR1 as shown in the amino acid sequence of SEQ ID NO: 1; The heavy chain CDR2 as shown in the amino acid sequence of SEQ ID NO: 2, wherein X=D or E; The heavy chain CDR3 as shown in the amino acid sequence of SEQ ID NO: 3; The light chain CDR1 as shown in the amino acid sequence of SEQ ID NO: 4; The light chain CDR2 as shown in the amino acid sequence of SEQ ID NO: 5; and The light chain CDR3 is shown in the amino acid sequence of SEQ ID NO:

6.

2. The anti-PD-L2 antibody or antigen-binding fragment thereof of claim 1, which is murine, chimeric or humanized.

3. The anti-PD-L2 antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein: (1) the heavy chain variable region and the light chain variable region comprise an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NOs: 25 and 26, respectively, (2) the heavy chain variable region and the light chain variable region comprise an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NOs: 33 and 34, respectively, or (3) the heavy chain variable region and the light chain variable region comprise an amino acid sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NOs: 35 and 34, respectively.

4. The anti-PD-L2 antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein: (1) the heavy chain variable region and the light chain variable region comprise the amino acid sequences shown in SEQ ID NOs: 25 and 26, respectively, (2) the heavy chain variable region and the light chain variable region comprise the amino acid sequences shown in SEQ ID NOs: 33 and 34, respectively, or (3) The heavy chain variable region and the light chain variable region comprise the amino acid sequences shown in SEQ ID NOs: 35 and 34, respectively.

5. The anti-PD-L2 antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, wherein: (1) the heavy chain and the light chain comprise an amino acid sequence that is at least 80% identical to the amino acid sequence shown in SEQ ID NOs: 46 and 47, respectively, (2) the heavy chain and the light chain comprise an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NOs: 62 and 63, respectively, or (3) The heavy chain and light chain comprise amino acid sequences that are at least 80% identical to the amino acid sequences shown in SEQ ID NOs: 64 and 63, respectively.

6. The anti-PD-L2 antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain and a light chain, wherein: (1) the heavy chain and the light chain comprise the amino acid sequences shown in SEQ ID NOs: 46 and 47, respectively, (2) the heavy chain and light chain comprise the amino acid sequences shown in SEQ ID NOs: 62 and 63, respectively, or (3) The heavy chain and light chain comprise the amino acid sequences shown in SEQ ID NOs: 64 and 63, respectively.

7. The anti-PD-L2 antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein the antibody or antigen-binding fragment thereof is selected from a monoclonal antibody, a Fab fragment, a F(ab')2 fragment, a Fv fragment, a single-chain Fv molecule or a combination thereof.

8. An isolated nucleic acid encoding the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

9. An expression vector comprising the nucleic acid according to claim 8. 10 . A host cell comprising the expression vector according to claim 9 .

11. A recombinant polypeptide comprising the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

12. A fusion protein comprising the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

13. A bispecific molecule comprising the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7.

14. A pharmaceutical composition comprising the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, and one or more pharmaceutically acceptable carriers.

15. The pharmaceutical composition of claim 14, wherein the pharmaceutical composition further comprises an anti-PD-L1 antibody.

16. Use of the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, or the pharmaceutical composition according to claim 14 or 15, in the preparation of a medicament for treating cancer in a subject, wherein the cancer is selected from: colorectal cancer, breast cancer, ovarian cancer, gastric cancer, kidney cancer, bladder cancer, head and neck cancer, liver cancer, small cell lung cancer, non-small cell lung cancer, melanoma, or a combination thereof.

17. A method for preparing the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7, the method comprising: (i) expressing the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 in a host cell, and (ii) isolating the anti-PD-L2 antibody or antigen-binding fragment thereof according to any one of claims 1 to 7 from the host cell or its cell culture.

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