Humanized anti-folate receptor 1 chimeric antigen receptor and its use

Abstract

The present invention describes a chimeric antigen receptor (CAR) specific for FOLR1, a vector encoding the FOLR1CAR, a recombinant host cell (CAR-T or CAR-NK) comprising the FOLR1CAR, and a method of using the CAR-T or CAR-NK to treat a disease associated with the expression of FOLR1. The present invention also describes a humanized anti-FOLR1 monoclonal antibody and an antigen-binding fragment thereof.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 62 / 832,975 filed on April 12, 2019, U.S. Provisional Application No. 62 / 863,330 filed on June 19, 2019, and U.S. Provisional Application No. 62 / 931,988 filed on November 7, 2019. The entire disclosure of each is incorporated herein by reference. Technical Field

[0003] The present invention relates to anti-folate receptor 1 (FOLR1) chimeric antigen receptor (CAR), nucleic acid encoding CAR and expression vector, T cells (CAR-T) engineered to express CAR and NK cells (CAR-NK) engineered to express CAR. Also provided are methods for preparing CAR, methods for preparing CAR-T / CAR-NK, and methods for using CAR-T / CAR-NK to treat diseases associated with FOLR1 expression, including cancer.

[0004] Reference to electronically submitted sequence listings

[0005] This application contains a sequence listing, which is submitted electronically via EFS-Web as an ASCII formatted sequence listing, with the file name "065799.21WO1 Sequence Listing" and a creation date of March 26, 2020, and a size of 140kb. The sequence listing submitted via EFS-Web is part of this specification, and the entire text of the sequence listing is incorporated herein by reference. Background Art

[0006] The standard of care for anticancer drugs provides significant benefits. Recently, the availability of immuno-oncology drugs such as anti-PD-1mAb, anti-PD-L1 mAb and anti-CD3 bispecific T cell adapters has advanced the concept of utilizing and activating the patient's immune system to fight various types of cancer. However, it is still necessary to solve the poor response, insufficient efficacy and / or safety issues. CAR-T (chimeric antigen receptor-T) cell therapy involves genetic engineering of the patient's own immune cells such as T cells, and redirecting them to suitable cell surface antigens on cancer cells (Mayor et al., Immunotherapy., 2016, Vol. 8, pp. 491 to 494). This approach has been proven to be successful in patients with chemo-refractory B cell malignancies and other cancers (Pettitt et al., Mol Ther., 2018, Vol. 26, pp. 342 to 353). T cells can be engineered to be specific to one or more cancer cell surface targets / antigens, so as to identify and kill cancer cells. The method includes transducing T cells with DNA or other genetic material encoding a chimeric antigen receptor (CAR), which comprises an extracellular antigen-specific binding domain, such as one or more single-chain variable fragments (scFv), hinge and transmembrane regions of a monoclonal antibody (mAb), and an intracellular signaling domain (including one or more co-stimulatory domains and one or more activation domains (Kochenderfer et al., Nat Rev Cl in Oncol., 2013, Vol. 10, pp. 267 to 276). Immune cells expressing CAR, such as T cells and NK cells, can be used to treat a variety of diseases, including liquid and solid tumors. Successful CAR-T cell therapy can specifically recognize and destroy target cells and maintain the ability to persist and proliferate over time.

[0007] Folate receptor 1 (FOLR1), also known as folate receptor α (FRα) or folate binding protein (FBP), is a glycosylphosphatidylinositol (GPI)-anchored membrane protein on the cell surface that has a high affinity for the active form of folate, 5-methyltetrahydrofolate (5-MTF) and its derivatives, and transports them into cells (Salazar and Ratnam, Cancer Metastasis Rev, 2007, Vol. 26, pp. 141-152). FOLR1 has become an oncology target because it is overexpressed in certain solid tumors such as ovarian cancer, lung cancer, and breast cancer (Toffoli et al., Int J Cancer, 1997, Vol. 74, pp. 193-198 and Boogerd et al., Oncotarget, 2016, Vol. 7, pp. 17442-17454), but is expressed at low levels in limited normal human tissues (Weitman et al., Cancer Res, 1992, Vol. 52, pp. 3396-3401). Consistent with this observation, phase 1 clinical trials conducted so far with small and large molecules targeting FOLR1 have revealed good drug tolerability (Cheung et al., Oncotarget, 2016, Vol. 7, pp. 52553-52574). Therefore, FOLR1 is an ideal target for CAR-T cell therapy to treat and cure FOLR1-positive cancers. Summary of the invention

[0008] In one general aspect, the present invention relates to a chimeric antigen receptor (CAR) construct that induces T cell-mediated cancer killing, wherein the CAR construct comprises at least one antigen binding domain that specifically binds to human folate receptor 1 (FOLR1), a hinge region, a transmembrane region, and an intracellular signaling domain.

[0009] Provided are isolated polynucleotides comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR). CAR may comprise (a) an extracellular domain comprising at least one antigen binding domain that specifically binds to folate receptor 1 (FOLR1); (b) a hinge region; (c) a transmembrane region; and (d) an intracellular signaling domain.

[0010] In certain embodiments, the antigen binding domain comprises a heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementary determining region 1 (LCDR1), LCDR2, and LCDR3, respectively having the following polypeptide sequences:

[0011] (1) SEQ ID NO: 17, 18, 19, 41, 42 and 43;

[0012] (2) SEQ ID NOs: 20, 21, 22, 44, 45 and 46;

[0013] (3) SEQ ID NOs: 23, 24, 25, 47, 48 and 49;

[0014] (4) SEQ ID NO: 26, 27, 28, 50, 51 and 52;

[0015] (5) SEQ ID NO: 29, 30, 31, 53, 54 and 55;

[0016] (6) SEQ ID NOs: 32, 33, 34, 56, 57 and 58;

[0017] (7) SEQ ID NO:35, 36, 37, 59, 60 and 61; or

[0018] (8) SEQ ID NO: 38, 39, 40, 62, 63 and 64;

[0019] Wherein the antigen binding domain specifically binds to FOLR1, preferably human FOLR1.

[0020] In certain embodiments, the antigen binding domain comprises a heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementary determining region 1 (LCDR1), LCDR2, and LCDR3, respectively having the following polypeptide sequences:

[0021] (1) SEQ ID NO: 65, 66, 67, 89, 90 and 91;

[0022] (2) SEQ ID NOs: 68, 69, 70, 92, 93 and 94;

[0023] (3) SEQ ID NO:71, 72, 73, 95, 96 and 97;

[0024] (4) SEQ ID NO:74, 75, 76, 98, 99 and 100;

[0025] (5) SEQ ID NO: 77, 78, 79, 101, 102 and 103;

[0026] (6) SEQ ID NOs: 80, 81, 82, 104, 105 and 106;

[0027] (7) SEQ ID NO: 83, 84, 85, 107, 108 and 109; or

[0028] (8) SEQ ID NO: 86, 87, 88, 110, 111 and 112;

[0029] Wherein the antigen binding domain specifically binds to FOLR1, preferably human FOLR1.

[0030] In certain embodiments, the antigen binding domain comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13 or 15, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or 16.

[0031] In certain embodiments, the antigen binding domain comprises:

[0032] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 2;

[0033] (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 3 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 4;

[0034] (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 5 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 6;

[0035] (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 8;

[0036] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 9 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 10;

[0037] (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 11 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 12;

[0038] (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14; or

[0039] (8) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0040] In certain embodiments, the antigen binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184, or having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: NO:117, 118, 121, 122, 123, 126, 127, 131, 132, 133, 134, 139, 140, 141, 145, 146, 147, 151, 152, 153, 157, 158 or 185-198 has a light chain variable region of a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical.

[0041] In certain embodiments, the antigen binding domain is humanized and comprises:

[0042] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0043] (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0044] (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0045] (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0046] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 117;

[0047] (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0048] (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0049] (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0050] (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0051] (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0052] (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0053] (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0054] (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0055] (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0056] (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 126;

[0057] (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 127;

[0058] (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0059] (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0060] (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0061] (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0062] (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0063] (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0064] (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0065] (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0066] (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0067] (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0068] (27) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0069] (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0070] (29) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0071] (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0072] (31) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0073] (32) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0074] (33) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0075] (34) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0076] (35) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0077] (36) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0078] (37) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0079] (38) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0080] (39) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0081] (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0082] (41) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0083] (42) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0084] (43) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0085] (44) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0086] (45) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0087] (46) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0088] (47) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151;

[0089] (48) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 150 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 153;

[0090] (49) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0091] (50) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0092] (51) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0093] (52) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0094] (53) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0095] (54) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0096] (55) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 182 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190;

[0097] (56) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0098] (57) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0099] (58) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197;

[0100] (59) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198;

[0101] (60) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0102] (61) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0103] (62) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197; or

[0104] (63) A heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198.

[0105] In certain embodiments, the antigen binding domain is a single chain variable fragment (scFv) that specifically binds FOLR1, preferably human FOLR1.

[0106] In certain embodiments, the antigen binding domain is a humanized single chain variable fragment (scFv) that specifically binds FOLR1, preferably human FOLR1. In certain embodiments, the humanized single chain variable fragment (scFv) comprises a polypeptide sequence that is at least 95% identical to any one of SEQ ID NOs: 159-170.

[0107] In certain embodiments, the chimeric antigen receptor (CAR) comprises one or more antigen binding domains.

[0108] In certain embodiments, the intracellular signaling domain comprises one or more costimulatory domains and one or more activation domains.

[0109] Also provided is a chimeric antigen receptor (CAR) encoded by the isolated polynucleotide of the invention.

[0110] Also provided is a vector comprising an isolated polynucleotide comprising a nucleic acid encoding a CAR of the present invention.

[0111] Also provided is a host cell comprising a vector of the invention.

[0112] In certain embodiments, the host cell is a T cell, preferably a human T cell. In certain embodiments, the host cell is a NK cell, preferably a human NK cell. T cells or NK cells can be, for example, engineered to express the CAR of the present invention to treat diseases such as cancer.

[0113] Also provided is a method for preparing a host cell expressing a chimeric antigen receptor (CAR) of the present invention. The method includes transducing T cells or NK cells with a vector comprising an isolated nucleic acid encoding the CAR of the present invention.

[0114] A method for producing the CAR-T cells or CAR-NK cells of the present invention is also provided. The method includes culturing T cells or NK cells comprising isolated polynucleotides under conditions of producing chimeric antigen receptor (CAR)-T cells or CAR-NK cells, and recovering the CAR-T cells or CAR-NK cells, wherein the isolated polynucleotides comprise nucleic acids encoding the CAR of the present invention.

[0115] Also provided is a method for generating an RNA engineered cell population comprising a chimeric antigen receptor (CAR) of the present invention. The method includes contacting a cell with a polynucleotide comprising a separation of a nucleic acid encoding a chimeric antigen receptor (CAR) of the present invention, wherein the polynucleotide of the separation is RNA or synthetic RNA transcribed in vitro.

[0116] Also provided is a method for treating a cancer of a subject in need thereof, the method comprising administering the CAR-T cells and / or CAR-NK cells of the present invention to a subject.Cancer can be any liquid or solid cancer, for example, it can be selected from but not limited to lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urethral cancer, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, and non-Hodgkin lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0117] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that increases the efficacy of cells expressing the CAR molecule.

[0118] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that improves one or more side effects associated with administering a cell expressing a CAR molecule.

[0119] In certain embodiments, the method of treating cancer in a subject in need thereof further comprises administering to the subject in need thereof an agent that treats a disease associated with FOLR1.

[0120] Also provided is a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to any one of SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184, or having a polypeptide sequence at least 95% identical to any one of SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184. NO:117, 118, 121, 122, 123, 126, 127, 131, 132, 133, 134, 139, 140, 141, 145, 146, 147, 151, 152, 153, 157, 158 or 185-198 has a light chain variable region of a polypeptide sequence that is at least 95% identical.

[0121] In certain embodiments, the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof comprises:

[0122] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0123] (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0124] (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0125] (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0126] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 117;

[0127] (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0128] (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0129] (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0130] (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0131] (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0132] (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0133] (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0134] (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0135] (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0136] (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 126;

[0137] (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 127;

[0138] (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0139] (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0140] (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0141] (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0142] (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0143] (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0144] (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0145] (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0146] (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0147] (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0148] (27) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0149] (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0150] (29) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0151] (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0152] (31) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0153] (32) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0154] (33) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0155] (34) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0156] (35) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0157] (36) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0158] (37) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0159] (38) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0160] (39) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0161] (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0162] (41) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0163] (42) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0164] (43) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0165] (44) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0166] (45) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0167] (46) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0168] (47) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151;

[0169] (48) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 150 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 153;

[0170] (49) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0171] (50) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0172] (51) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0173] (52) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0174] (53) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0175] (54) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0176] (55) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 182 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190;

[0177] (56) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0178] (57) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0179] (58) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197;

[0180] (59) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198;

[0181] (60) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0182] (61) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0183] (62) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197; or

[0184] (63) A heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198.

[0185] In certain embodiments, the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof is capable of binding to FOLR1, inducing effector-mediated tumor cell lysis, mediating the recruitment of conjugated drugs, and / or forming a bispecific antibody with another monoclonal antibody or antigen-binding fragment having cancer killing effects.

[0186] Also provided are isolated nucleic acids encoding the humanized anti-FOLR1 monoclonal antibodies or antigen-binding fragments thereof of the invention.

[0187] Also provided are vectors comprising an isolated nucleic acid encoding a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the invention.

[0188] Also provided is a host cell comprising a vector comprising an isolated nucleic acid encoding a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the invention.

[0189] Also provided is a pharmaceutical composition comprising the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the present invention and a pharmaceutically acceptable carrier.

[0190] Also provided is a method of targeting FOLR1 on the surface of cancer cells in a subject in need thereof, the method comprising administering to the subject in need thereof a pharmaceutical composition comprising the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the present invention.

[0191] Also provided is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof of the present invention. The cancer can be any liquid or solid cancer, for example, it can be selected from but not limited to lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0192] Also provided is a method for producing a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof of the present invention, the method comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions in which the monoclonal antibody or antigen-binding fragment is produced, and recovering the antibody or antigen-binding fragment from the cell or culture.

[0193] Also provided is a method for producing a pharmaceutical composition comprising the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the present invention, the method comprising combining the monoclonal antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0194] The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the accompanying drawings. It should be understood, however, that the present application is not limited to the precise embodiments shown in the drawings.

[0195] Figures 1A to 1L Shown is the binding of humanized mAbs to immobilized recombinant human FOLR1 protein as determined by ELISA.

[0196] Figures 2A to 2E Binding of humanized mAbs to SK-OV-3 cells is shown. The experiment was performed by FACS analysis.

[0197] Figures 3A to 3G Shown is the binding of humanized scFv to immobilized recombinant human FOLR1 protein as determined by ELISA.

[0198] Figures 4A to 4G Binding of humanized scFv to SK-OV-3 cells is shown. The experiment was performed by FACS analysis. DETAILED DESCRIPTION

[0199] Various publications, articles, and patents are cited or described in the Background and throughout the specification; each of these references is incorporated herein by reference in its entirety. The discussion of documents, acts, materials, devices, articles, etc. included in this specification is intended to provide context for the present invention. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any invention disclosed or claimed.

[0200] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Otherwise, certain terms used herein have the meanings described in this specification.

[0201] It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0202] Unless otherwise indicated, any numerical value, such as a concentration or concentration range described herein, is understood to be modified by the term "about" in all cases. Therefore, numerical values ​​generally include ±10% of the listed value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL to 1.1 mg / mL. Similarly, a concentration range of 1% to 10% (w / v) includes 0.9% (w / v) to 11% (w / v). As used herein, unless the context clearly indicates otherwise, the use of numerical ranges clearly includes all possible subranges, all individual values ​​within the range, including integers and fractions of values ​​within such ranges.

[0203] Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to each element in the series. Those skilled in the art will recognize or be able to ascertain, using only routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0204] As used herein, the terms "comprises, comprising," "includes, including," "has, having," "contains or containing," or any other variation thereof, will be understood to imply the inclusion of the specified whole or group of wholes, but not the exclusion of any other whole or group of wholes, and are intended to be non-exclusive or open-ended. For example, a composition, mixture, process, method, article, or apparatus that includes a series of elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or apparatus. In addition, unless expressly indicated to the contrary, "or" refers to an inclusive "or" rather than an exclusive "or." For example, a condition A or B satisfies any of the following conditions: A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), and both A and B are true (or exist).

[0205] As used herein, the connectivity term "and / or" between multiple listed elements should be understood to cover both single and combined options. For example, where two elements are connected by "and / or", the first option refers to the applicability of the first element without the second element. The second option refers to the applicability of the second element without the first element. The third option refers to the applicability of the first element and the second element together. Any of these options is understood to fall within this meaning and therefore meets the requirements of the term "and / or" as used herein. The parallel applicability of more than one option is also understood to fall within this meaning and therefore meets the requirements of the term "and / or".

[0206] As used herein, the term "consists of" or variations such as "consist of or consisting of" as used throughout the specification and claims means that any listed integer or group of integers is included, but additional integers or groups of integers cannot be added to a specified method, structure or composition.

[0207] As used herein, the term "consists essentially of" or variations thereof such as "consist essentially of or consisting essentially of" as used throughout the specification and claims means including any recited integer or group of integers, and optionally including any recited integer or group of integers that does not materially alter the basic or novel characteristics of a specified method, structure, or composition. See MPEP §2111.03.

[0208] As used herein, "subject" means any animal, preferably a mammal, most preferably a human. As used herein, the term "mammal" encompasses any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., more preferably humans.

[0209] The words "right," "left," "lower," and "upper" designate directions in the drawings to which reference is made.

[0210] It should also be understood that when referring to dimensions or characteristics of components of the preferred invention, the use of "about," "approximately," "substantially," and similar terms herein indicates that the described dimensions / characteristics are not strict boundaries or parameters and do not exclude minor variations that are functionally the same or similar, as understood by those of ordinary skill in the art. At a minimum, such references including numerical parameters will include variations that do not change the least significant digit using mathematical and industrial principles accepted in the art (e.g., rounding, measurement or other systematic errors, manufacturing tolerances, etc.).

[0211] The terms "identity" or "percent identity" in the context of two or more nucleic acid or polypeptide sequences (e.g., chimeric antigen receptors (CARs) comprising antigen binding domains specific for FOLR1 and polynucleotides encoding them, FOLR1 polypeptides, and FOLR1 polynucleotides encoding them) refer to two or more sequences or subsequences that are identical or have a specified percentage of amino acid residues or nucleotides that are identical when compared and aligned for maximum correspondence using one of the following sequence comparison algorithms or by visual inspection.

[0212] For sequence comparison, usually one sequence is used as a reference sequence to which a test sequence is compared. When a sequence comparison algorithm is used, the test sequence and the reference sequence are input into a computer, subsequence coordinates are specified if necessary, and sequence algorithm program parameters are specified. The sequence comparison algorithm then calculates the sequence identity percentage of the test sequence relative to the reference sequence based on the specified program parameters.

[0213] Optimal alignment of sequences for comparison can be achieved, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math., 1981, volume 2, page 482, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol., 1970, volume 48, page 443, by the similarity search method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA, 1988, volume 85, page 2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Avenue, Madison, Wisconsin), or by visual inspection (see generally "Current Protocols in Molecular Biology", FM Ausubel et al., eds., Current Protocols in Molecular Biology). Protocols, a joint venture between Green Publishing Associates and John Wiley & Sons, 1995 Supplement (Ausubel).

[0214] Examples of algorithms suitable for determining percentages of sequence identity and sequence similarity are BLAST and BLAST2.0 algorithms, which are described in Altschul et al., J.Mol.Biol., 1990, Vol. 215, pp. 403 to 410 and Altschul et al., Nucleic Acids Res., 1997, Vol. 25, pp. 3389 to 3402. Software for BLAST analysis is publicly available through the National Center for Biotechnology Information. The algorithm includes first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which match or satisfy some positive threshold score T when compared with words of the same length in the database sequence. T is referred to as the contiguous word score threshold (Altschul et al., supra). These initial contiguous word hits are used as seeds for initiating searches to find longer HSPs containing them. Then, as long as the cumulative comparison score can be increased, the word hits are extended in both directions along each sequence.

[0215] For nucleotide sequences, the cumulative score is calculated using the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatched residues; always < 0). For amino acid sequences, the cumulative score is calculated using a scoring matrix. The extension of the word hit in each direction stops when the following occurs: the cumulative alignment score drops by the amount X from its maximum achieved value; the cumulative score becomes zero or lower due to the accumulation of one or more negative scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses a word length (W) of 11, an expectation (E) of 10, M = 5, N = -4, and a comparison of two chains by default. For amino acid sequences, the BLASTP program uses a word length (W) of 3, an expectation (E) of 10, and a BLOSUM62 scoring matrix by default (see Henikoff & Henikoff, Proc. Nat'l. Acad. Sci. USA, 1989, Vol. 89, p. 10915).

[0216] In addition to calculating the percentage of sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA, 1993, Vol. 90, pp. 5873 to 5787). A similarity measure provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability that a match occurs by chance between two nucleotide or amino acid sequences. For example, if the smallest sum probability is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001 in a comparison of a test nucleic acid to a reference nucleic acid, the nucleic acid is considered similar to the reference sequence.

[0217] Another indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, where the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions.

[0218] As used herein, the term "isolated" means that a biological component (such as a nucleic acid, peptide or protein) has been substantially separated, produced separately or purified from other biological components of the organism in which the component naturally occurs, i.e., other chromosomal and extrachromosomal DNA and RNA and proteins. Thus, nucleic acids, peptides and proteins that have been "isolated" include nucleic acids and proteins purified by standard purification methods. "Isolated" nucleic acids, peptides and proteins can be part of a composition and can still be isolated if the composition is not part of the native environment of the nucleic acid, peptide or protein. The term also includes nucleic acids, peptides and proteins prepared by recombinant expression in a host cell and chemically synthesized nucleic acids.

[0219] As used herein, the term "polynucleotide", synonymously referred to as "nucleic acid molecule", "nucleotide" or "nucleic acid", refers to any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. "Polynucleotide" includes, but is not limited to, single-stranded and double-stranded DNA, DNA mixed with single-stranded and double-stranded regions, single-stranded and double-stranded RNA, RNA mixed with single-stranded and double-stranded regions, hybrid molecules containing DNA and RNA that may be single-stranded or more typically double-stranded or mixed with single-stranded and double-stranded regions. In addition, "polynucleotide" refers to a triple-stranded region containing RNA or DNA or both RNA and DNA. The term "polynucleotide" also includes DNA or RNA containing one or more modified bases, as well as DNA or RNA whose backbone is modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and rare bases such as inosine. DNA and RNA can be modified in various ways; therefore, "polynucleotide" includes chemical, enzymatic or metabolic modified forms of polynucleotides that are usually naturally occurring, as well as chemical forms of DNA and RNA that are unique to viruses and cells. "Polynucleotide" also includes relatively short nucleic acid chains, often referred to as oligonucleotides.

[0220] As used herein, the term "vector" is a replicon into which another nucleic acid segment may be operably inserted so as to bring about the replication or expression of that segment.

[0221] As used herein, the term "host cell" refers to a cell comprising a nucleic acid molecule of the present invention. A "host cell" can be any type of cell, for example, a primary cell, a cultured cell, or a cell from a cell line. In one embodiment, a "host cell" is a cell transfected or transduced with a nucleic acid molecule of the present invention. In another embodiment, a "host cell" is a progeny or potential progeny of such a transfected or transduced cell. The progeny of a cell may be identical to or different from the parent cell, for example, due to mutations or environmental influences that may occur in subsequent generations, or the integration of nucleic acid molecules into the host cell genome.

[0222] As used herein, the term "expression" refers to the biosynthesis of a gene product. The term encompasses the transcription of a gene to RNA. The term also encompasses the translation of RNA to one or more polypeptides, and also encompasses all naturally occurring post-transcriptional and post-translational modifications. The expressed CAR may be in the cytoplasm of a host cell, in an extracellular environment such as a growth medium for a cell culture, or anchored to a cell membrane.

[0223] As used herein, the term "immune cell" or "immune effector cell" refers to a cell that participates in an immune response, such as a cell that promotes an immune effector response. Examples of immune cells include T cells, B cells, natural killer (NK) cells, mast cells, and bone marrow-derived phagocytes. According to a specific embodiment, the engineered immune cells are T cells and are referred to as CAR-T cells because they are engineered to express the CAR of the present invention.

[0224] As used herein, the term "engineered immune cell" refers to an immune cell that has been genetically modified by adding additional genetic material in the form of DNA or RNA to the total genetic material of the cell, also referred to as an immune effector cell. According to embodiments herein, engineered immune cells have been genetically modified to express a CAR construct according to the present invention.

[0225] Chimeric Antigen Receptor (CAR)

[0226] As used herein, the term "chimeric antigen receptor" (CAR) refers to a recombinant polypeptide comprising an extracellular domain, a transmembrane domain, and an intracellular T cell receptor activation signaling domain that specifically binds at least an antigen or target. The engagement of the extracellular domain of CAR with the target antigen on the surface of the target cell results in the clustering of CAR, and delivers activation stimulation to cells containing CAR. CAR redirects the specificity of immune effector cells and triggers proliferation, cytokine production, phagocytosis, and / or produces molecules that can mediate cell death of target antigen-expressing cells in a major histocompatibility (MHC)-independent manner.

[0227] In one aspect, CAR comprises an antigen binding domain, a hinge region, a costimulatory domain, an activation domain, and a transmembrane region. In one aspect, CAR comprises an antigen binding domain, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region. In one aspect, CAR comprises two antigen binding domains, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region. In one aspect, CAR comprises two antigen binding domains, a hinge region, two costimulatory domains, an activation domain, and a transmembrane region.

[0228] As used herein, the term "signal peptide" refers to a leader sequence at the amino terminus (N-terminus) of a nascent CAR protein that co-translationally or post-translationally directs the nascent protein to the endoplasmic reticulum and subsequently to surface expression.

[0229] As used herein, the term "extracellular antigen binding domain", "extracellular domain" or "extracellular ligand binding domain" refers to the portion of CAR that is located outside the cell membrane and is capable of binding to an antigen, target or ligand.

[0230] As used herein, the term "hinge region" refers to the portion of a CAR that connects two adjacent domains of a CAR protein, such as an extracellular domain and a transmembrane domain.

[0231] As used herein, the term "transmembrane domain" refers to the portion of CAR that extends across the cell membrane and anchors CAR to the cell membrane. It is sometimes referred to as a "transmembrane region".

[0232] Costimulatory domain

[0233] As used herein, chimeric antigen receptors can be incorporated with costimulatory (signaling) domains to increase their effectiveness. Costimulatory (signaling) domains can be derived from costimulatory molecules. Costimulatory molecules are cell surface molecules other than antigen receptors or their ligands required for effective immune responses. Costimulatory domains can be derived from costimulatory molecules, which may include but are not limited to CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (α, β, δ, ε, γ, ζ), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, programmed death receptor-1 (PD-1), inducible T cell costimulators (ICOS), lymphocyte function-associated antigen-1 (LFA-1; CD 11a and CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Igα (CD79a), DAP10, fcγ receptor, MHC class I molecule, TNFR, integrin, signaling lymphocyte activation molecule, BTLA, Toll ligand receptor, ICAM-1, CDS, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD1 9. CD8α, CD8β, IL-2Rβ, IL-2Rγ, IL-7Rα, ITGA4, VLA1, CD49a, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, ITGAE, CD103, ITGAL, CD1a, CD1 b, CD1c, CD1d, ITGAM, ITGAX, ITGB1, CD29, ITGB2(CD18), ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD 84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand, cytokine receptor, activating NK cell receptor or their fragments or any combination.

[0234] Activation domain

[0235] As used herein, a chimeric antigen receptor may comprise an activation domain. The activation domain may include, but is not limited to, CD3. CD3 is an element of the T cell receptor on natural T cells and has been shown to be an important intracellular activation element in CAR. In a preferred embodiment, CD3 is CD3ζ.

[0236] Hinge area

[0237] As described herein, chimeric antigen receptors may include a hinge region. This is a portion of the extracellular domain, sometimes referred to as a "spacer" region. A variety of hinges may be used according to the present invention, including costimulatory molecules, immunoglobulin (Ig) sequences or other suitable molecules as described above, to achieve a desired specific distance from a target cell. In some embodiments, the entire extracellular region includes a hinge region.

[0238] Transmembrane region

[0239] As used herein, chimeric antigen receptor (CAR) may include a transmembrane region / domain. CAR may be designed to include a transmembrane domain fused to the extracellular domain of CAR. It may be similarly fused to the intracellular domain of CAR. In one embodiment, a transmembrane domain naturally associated with a domain in the domain in CAR is used. In some cases, the transmembrane domain may be selected or modified by amino acid substitution to avoid such domains from binding to the transmembrane domain of the same or different surface membrane proteins, thereby minimizing the interaction with other members of the receptor complex. The transmembrane domain may be derived from a natural source or a synthetic source. In the case where the source is natural, the domain may be derived from any membrane-bound protein or transmembrane protein.The transmembrane region specifically used in the present invention may be derived from (i.e., include or engineered from) but is not limited to CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (α, β, δ, ε, γ, ζ), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, programmed death receptor-1 (PD-1), inducible T cell co-stimulator (ICOS), lymphocyte function-associated antigen-1 (LFA-1; CD11a and CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Igα (CD79a), DAP10, fcγ receptor, MHC class I molecule, TNFR, integrin, signaling lymphocyte activation molecule, BTLA, Toll ligand receptor, ICAM-1, CDS, GITR, BAFFR, LIGHT, HVEM(LIGHTR), KIRDS2, SLAMF7, NKp80(KLRF1), NKp44, NKp30, NKp46, CD 19. CD8α, CD8β, IL-2Rβ, IL-2Rγ, IL-7Rα, ITGA4, VLA1, CD49a, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, ITGA E. CD103, ITGAL, CD1a, CD1b, CD1c, CD1d, ITGAM, ITGAX, ITGB1, CD29, ITGB2(CD18), ITGB7, NKG2D, TNFR2, T RANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRTAM, Ly9(CD229), CD 160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand, cytokine receptor, activating NK cell receptor, immunoglobulin protein or fragments thereof or any combination thereof.

[0240] Immune cells

[0241] According to a specific aspect, the present invention provides a cell as an immune cell, comprising a separated polynucleotide or a vector comprising a separated polynucleotide, the separated polynucleotide comprising a nucleotide sequence encoding CAR provided herein. Immune cells comprising the separated polynucleotides and / or vectors of the present invention may be referred to as "engineered immune cells". Preferably, engineered immune cells are derived from humans (human-derived before being made into recombinant).

[0242] For example, the engineered immune cell can be a cell of the lymphoid lineage. Non-limiting examples of cells of the lymphoid lineage can include T cells and natural killer (NK) cells. T cells express T cell receptors (TCRs), with most cells expressing gamma and delta chains, and smaller groups expressing gamma and delta chains. The T cells used as engineered immune cells of the present invention can be CD4 + or CD8 + , and may include but are not limited to T helper cells (CD4 + ), cytotoxic T cells (also called cytotoxic T lymphocytes, CTL; CD8 + Cells) and memory T cells, including central memory T cells, stem cell-like memory T cells and effector memory T cells, natural killer T cells, mucosal-associated invariant T cells and γδT cells.Other exemplary immune cells include, but are not limited to, macrophages, antigen presenting cells (APCs), or any immune cells expressing inhibitors of cell-mediated immune responses such as immune checkpoint inhibitor pathway receptors (e.g., PD-1). Precursor cells of immune cells that can be used according to the present invention include hematopoietic stem cells and / or progenitor cells.Hematopoietic stem cells and / or progenitor cells can be derived from bone marrow, umbilical cord blood, adult peripheral blood after cytokine mobilization, etc., by methods known in the art.Immune cells are engineered to recombinantly express the CAR of the present invention.

[0243] Immune cells and their precursor cells can be separated by methods known in the art, including commercially available methods (see, for example, Rowland Jones et al., Lymphocytes: A Practical Approach, Oxford University Press, New York, 1999). The sources of immune cells or their precursors include, but are not limited to, peripheral blood, cord blood, bone marrow, or other hematopoietic cell sources. Various techniques can be used to separate cells to separate or enrich the desired immune cells. For example, negative selection methods can be used to remove cells that are not desired immune cells. In addition, positive selection methods can be used to separate or enrich the desired immune cells or their precursors, or a combination of positive selection methods and negative selection methods can be used. If a specific type of cell is to be separated, such as a specific T cell, various cell surface markers or a combination of markers (e.g., CD3, CD4, CD8, CD34) can be used to separate cells.

[0244] Immune cells or their precursor cells can be autologous or non-autologous for the subjects to whom they are applied in the treatment methods of the present invention. Autologous cells are isolated from subjects to be administered engineered immune cells that recombinantly express CAR. Optionally, cells can be obtained by leukocyte separation, in which leukocytes are selectively removed from the drawn blood, recombinants are made, and then transferred to the donor. Alternatively, allogeneic cells from non-autologous donors of non-subjects can be used. In the case of non-autologous donors, cells are typed and matched with human leukocyte antigens (HLA) to determine the appropriate level of compatibility. For autologous cells and non-autologous cells, cells can be optionally cryopreserved until ready for use.

[0245] Various methods for isolating immune cells that can be used to recombinantly express the CAR of the present invention have been previously described and can be used, including but not limited to the use of peripheral donor lymphocytes (Sadelain et al., Nat. Rev. Cancer, 2003, Vol. 3, pp. 35-45; Morgan et al., Science, 2006, Vol. 314, pp. 126-129), the use of lymphocyte cultures derived from tumor infiltrating lymphocytes (TIL) in tumor biopsies (Panelli et al., J. Immunol., 2000, Vol. 164, pp. 495-504; Panelli et al., J. Immunol., 2000, Vol. 164, pp. 4382-4392), and the use of antigen-specific peripheral blood leukocytes selectively expanded in vitro using artificial antigen presenting cells (AAPCs) or dendritic cells (Dupont et al., Cancer Res, 2005, Vol. 65, pp. 5417-5427; Papanicolaou et al., Blood, 2003, Vol. 102, pp. 2498-2505). Where stem cells are used, the cells can be isolated by methods well known in the art (see, e.g., Klug et al., Hematopoietic Stem Cell Protocols, Humana Press, New York, 2002; Freshney et al., Culture of Human Stem Cells, John Wiley & Sons, 2007).

[0246] According to a specific embodiment, the method for preparing engineered immune cells includes transfection or transduction of immune effector cells separated from individuals, so that immune effector cells express one or more CARs according to embodiments of the present invention. The method for preparing immune cells for immunotherapy is described in, for example, WO2014 / 130635, WO2013 / 176916, and WO2013 / 176915, and these documents are incorporated herein by reference. The various steps that can be used to prepare engineered immune cells are disclosed in, for example, WO2014 / 039523, WO2014 / 184741, WO2014 / 191128, WO2014 / 184744, and WO2014 / 184143, and these documents are incorporated herein by reference.

[0247] In a specific embodiment, immune effector cells, such as T cells, are genetically modified (for example, transduced with a viral vector comprising a nucleic acid encoding CAR) with the CAR of the present invention, and then activated and amplified in vitro. In various embodiments, before or after the genetic modification of expressing CAR, the method described in, for example, US6352694, US6534055, US6905680, US6692964, US5858358, US6887466, US6905681, US7144575, US7067318, US7172869, US7232566, US7175843, US5883223, US6905874, US6797514, US6867041, US2006 / 121005 is used to activate and amplify T cells, which are incorporated herein by reference. T cells can be amplified in vitro or in vivo. Typically, the T cells of the present invention can be amplified by contact with a surface, which is attached with agents that stimulate CD3 / TCR complex-related signals and ligands for stimulating co-stimulatory molecules on the surface of T cells. As a non-limiting example, T cell groups can be stimulated as described herein, such as by contacting with anti-CD3 antibodies or their antigen-binding fragments or anti-CD3 antibodies fixed on the surface, or by contacting with protein kinase C activators (e.g., bryostatin) bound to calcium ion carriers, or by activation of CAR itself. For the co-stimulation of auxiliary molecules on the surface of T cells, ligands for binding auxiliary molecules are used. For example, T cell groups can be contacted with anti-CD3 antibodies and anti-CD28 antibodies under conditions suitable for stimulating T cell proliferation. Conditions suitable for T cell culture include, for example, factors necessary for proliferation and survival, including serum (e.g., fetal bovine serum or human serum), cytokines such as IL-2, IL-7, IL-15 and / or IL-21, insulin, IFN-g, GM-CSF, TGFβ and / or any other additives known to those skilled in the art for cell growth in an appropriate culture medium (e.g., minimal essential medium or RPMI culture medium 1640 or X-Vivo 5 (Lonza)). In other embodiments, feeder cells and appropriate antibodies and cytokines may be used to activate and stimulate T cell proliferation, such as methods described in US6040177, US5827642 and WO2012129514, which are incorporated herein by reference.

[0248] Antibodies and antigen binding domains

[0249] As used herein, the term "antibody" is used in its broad sense, including immunoglobulins or antibody molecules, including human antibodies, humanized antibodies, composite antibodies and chimeric antibodies and monoclonal or polyclonal antibody fragments. Generally speaking, antibodies are proteins or peptide chains that show binding specificity to specific antigens. Antibody structure is well known. According to the heavy chain constant domain amino acid sequence, immunoglobulins can be designated as five major categories (i.e., IgA, IgD, IgE, IgG and IgM). IgA and IgG are further subdivided into isotypes IgA1, IgA2, IgG1, IgG2, IgG3 and IgG4. Therefore, the antibody of the present invention can be any of the five major categories or corresponding subtypes. Preferably, the antibody of the present invention is IgG1, IgG2, IgG3 or IgG4. The antibody light chain of a vertebrate species can be designated as one of two completely different types, i.e., κ and λ, based on the amino acid sequence of its constant domain. Therefore, the antibody of the present invention can contain a κ light chain constant domain or a λ light chain constant domain. According to a specific embodiment, the antibody of the present invention comprises a heavy chain and / or light chain constant region from a rat or human antibody. In addition to the heavy chain and light chain constant domains, the antibody also contains an antigen binding region consisting of a light chain variable region and a heavy chain variable region, wherein each variable region contains three domains (i.e., complementary determining regions 1 to 3; CDR1, CDR2, and CDR3). The light chain variable region domain is alternatively referred to as LCDR1, LCDR2, and LCDR3, and the heavy chain variable region domain is alternatively referred to as HCDR1, HCDR2, and HCDR3.

[0250] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigenic specificities (e.g., an isolated antibody that specifically binds FOLR1 is substantially free of antibodies that do not bind FOLR1). In addition, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0251] As used herein, the term "monoclonal antibody" or "mAb" refers to an antibody obtained from a substantially homogeneous antibody population, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in small amounts. The monoclonal antibodies of the present invention can be prepared by hybridoma methods, phage display technology, single lymphocyte gene cloning technology, or by recombinant DNA methods. For example, monoclonal antibodies can be produced by a hybridoma comprising a B cell obtained from a transgenic non-human animal such as a transgenic mouse or rat having a genome comprising a human heavy chain transgene and a light chain transgene.

[0252] As used herein, the terms "antigen binding fragment" and / or "antigen binding domain" refer to antibody fragments, such as, for example, diabodies, Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized diabodies (dsdiabodies), single-chain antibody molecules (scFv), single domain antibodies (sdab) and scFv dimers (bivalent diabodies), multispecific antibodies formed from a portion of an antibody comprising one or more CDRs, camelized single domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment that binds to an antigen but does not contain a complete antibody structure. The antigen binding domain is capable of binding to the same antigen that the parent antibody binds to. According to a specific embodiment, the antigen binding domain comprises a single-chain antibody molecule (scFv).

[0253] As used herein, the term "single-chain antibody" refers to a conventional single-chain antibody in the art, which comprises a heavy chain variable region and a light chain variable region connected by a short peptide of about 5 to about 20 amino acids. As used herein, the term "single domain antibody" refers to a conventional single domain antibody in the art, which comprises a heavy chain variable region and a heavy chain constant region or only a heavy chain variable region.

[0254] As used herein, the term "human antibody" refers to an antibody produced by a human or an antibody prepared using any technique known in the art having an amino acid sequence corresponding to an antibody produced by a human. This definition of a human antibody includes complete or full-length antibodies, fragments thereof, and / or antibodies comprising at least one human heavy chain polypeptide and / or light chain polypeptide.

[0255] As used herein, the terms "humanized antibody" and / or "humanized antigen-binding domain" refer to non-human antibodies and / or non-human antigen-binding domains that have been modified to increase sequence homology with human antibodies and / or human antigen-binding domains, such that the antigen-binding properties of the antigen-binding domain are retained, but its antigenicity in humans is reduced.

[0256] As used herein, the term "chimeric antibody" and / or "chimeric antigen-binding domain" refers to an antibody and / or antigen-binding domain in which the amino acid sequence of an immunoglobulin molecule is derived from two or more species. The variable regions of both the light and heavy chains generally correspond to the variable regions of an antibody and / or antigen-binding domain with the desired specificity, affinity, and capacity derived from one mammalian (e.g., mouse, rat, rabbit, etc.) species, while the constant region corresponds to the sequence of an antibody and / or antigen-binding domain derived from another mammalian (e.g., human) species to avoid eliciting an immune response in that species.

[0257] As used herein, the term "multi-specific antibody" refers to an antibody comprising multiple immunoglobulin variable domain sequences, wherein the first immunoglobulin variable domain sequence in the multiple immunoglobulin variable domain sequences has binding specificity to the first epitope, and the second immunoglobulin variable domain sequence in the multiple immunoglobulin variable domain sequences has binding specificity to the second epitope. In one embodiment, the first epitope and the second epitope are located on the same antigen, for example, on the same protein (or subunit of a multimeric protein). In one embodiment, the first epitope and the second epitope overlap or substantially overlap. In one embodiment, the first epitope and the second epitope do not overlap or substantially do not overlap. In one embodiment, the first epitope and the second epitope are located on different antigens, for example, on different proteins (or different subunits of a multimeric protein). In one embodiment, the multi-specific antibody comprises a third immunoglobulin variable domain, a fourth immunoglobulin variable domain, or a fifth immunoglobulin variable domain. In one embodiment, the multi-specific antibody is a bispecific antibody molecule, a tri-specific antibody molecule, or a tetra-specific antibody molecule.

[0258] As used herein, the term "bispecific antibody" refers to a multispecific antibody that binds no more than two epitopes or two antigens. The bispecific antibody is characterized by a first immunoglobulin variable domain sequence with binding specificity to a first epitope and a second immunoglobulin variable domain sequence with binding specificity to a second epitope. In one embodiment, the first epitope and the second epitope are located on the same antigen, for example, on the same protein (or subunit of a multimeric protein). In one embodiment, the first epitope and the second epitope overlap or substantially overlap. In one embodiment, the first epitope and the second epitope are located on different antigens, for example, on different proteins (or different subunits of a multimeric protein). In one embodiment, the bispecific antibody comprises a heavy chain variable domain sequence and a light chain variable domain sequence with binding specificity to the first epitope and a heavy chain variable domain sequence and a light chain variable domain sequence with binding specificity to the second epitope. In one embodiment, the bispecific antibody comprises a half antibody or a fragment thereof with binding specificity to the first epitope and a half antibody or a fragment thereof with binding specificity to the second epitope. In one embodiment, the bispecific antibody comprises a scFv or fragment thereof having binding specificity to a first epitope and a scFv or fragment thereof having binding specificity to a second epitope. In one embodiment, the first epitope is located on FOLR1, and the second epitope is located on PD-1, PD-L1, TIM-3, LAG-3, CD73, apelin, CTLA-4, EGFR, HER-2, CD3, CD19, CD20, CD33, CD47, TIP-1, CLDN18.2, DLL3 and / or other tumor-associated immunosuppressive factors or surface antigens.

[0259] As used herein, the term "FOLR1" refers to folate receptor 1 (FOLR1), also known as folate receptor α (FRα) or folate binding protein (FBP), which is a glycosylphosphatidylinositol (GPI)-anchored membrane protein on the cell surface that has a high affinity for the active form of folate, 5-methyltetrahydrofolate (5-MTF) and its derivatives, and transports them into cells (Salazar and Ratnam, Cancer Metastasis Rev, 2007, Vol. 26, pp. 141-152). FOLR1 has become an oncology target because it is overexpressed in certain solid tumors such as ovarian cancer, lung cancer, and breast cancer (Toffoli et al., Int J Cancer, 1997, Vol. 74, pp. 193-198 and Boogerd et al., Oncotarget, 2016, Vol. 7, pp. 17442-17454), but is expressed at low levels in limited normal human tissues (Weitman et al., Cancer Res, 1992, Vol. 52, pp. 3396-3401). Consistent with this observation, phase 1 clinical trials conducted so far with small and large molecules targeting FOLR1 have revealed good drug tolerability (Cheung et al., Oncotarget, 2016, Vol. 7, pp. 52553-52574). Therefore, FOLR1 is an ideal target for CAR-T cell therapy to treat and cure FOLR1-positive cancers. An exemplary amino acid sequence of human FOLR1 is represented by GenBank Accession No. NP_057937 (SEQ ID NO: 135).

[0260] As used herein, an antibody and / or antigen binding domain that "specifically binds to FOLR1" refers to an antibody and / or antigen binding domain that specifically binds to FOLR1 at a rate of 1×10 -7 M or less, preferably 1×10 -8 M or less, more preferably 5×10 -9 M or less, 1×10 -9 M or lower, 5×10 -10 M or less or 1×10 -10 The term "KD" refers to the 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 the antigen binding domain can be determined according to the present disclosure using methods in the art. For example, it can be determined by using surface plasmon resonance, such as by using a biosensor system, such as systems, or by using bio-layer interferometry techniques, such as the Octet RED96 system, to determine the KD of antibodies and / or antigen binding domains.

[0261] The smaller the KD value of the antibody and / or antigen-binding domain, the higher the affinity of the antibody and / or antigen-binding domain for binding to the target antigen.

[0262] According to a specific aspect, the present invention relates to a chimeric antigen receptor (CAR) comprising an antigen binding domain, wherein the antigen binding domain comprises a heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementary determining region 1 (LCDR1), LCDR2 and LCDR3, respectively having the following polypeptide sequences:

[0263] (1) SEQ ID NO: 17, 18, 19, 41, 42 and 43;

[0264] (2) SEQ ID NOs: 20, 21, 22, 44, 45 and 46;

[0265] (3) SEQ ID NOs: 23, 24, 25, 47, 48 and 49;

[0266] (4) SEQ ID NO: 26, 27, 28, 50, 51 and 52;

[0267] (5) SEQ ID NO: 29, 30, 31, 53, 54 and 55;

[0268] (6) SEQ ID NOs: 32, 33, 34, 56, 57 and 58;

[0269] (7) SEQ ID NO:35, 36, 37, 59, 60 and 61; or

[0270] (8) SEQ ID NO: 38, 39, 40, 62, 63 and 64;

[0271] Wherein the antigen binding domain specifically binds to FOLR1, preferably human FOLR1.

[0272] According to another specific aspect, the present invention relates to a chimeric antigen receptor (CAR) comprising an antigen binding domain, wherein the antigen binding domain comprises a heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementary determining region 1 (LCDR1), LCDR2 and LCDR3, respectively having the following polypeptide sequences:

[0273] (1) SEQ ID NO: 65, 66, 67, 89, 90 and 91;

[0274] (2) SEQ ID NOs: 68, 69, 70, 92, 93 and 94;

[0275] (3) SEQ ID NO:71, 72, 73, 95, 96 and 97;

[0276] (4) SEQ ID NO:74, 75, 76, 98, 99 and 100;

[0277] (5) SEQ ID NO: 77, 78, 79, 101, 102 and 103;

[0278] (6) SEQ ID NOs: 80, 81, 82, 104, 105 and 106;

[0279] (7) SEQ ID NO: 83, 84, 85, 107, 108 and 109; or

[0280] (8) SEQ ID NO: 86, 87, 88, 110, 111 and 112;

[0281] Wherein the antigen binding domain specifically binds to FOLR1, preferably human FOLR1.

[0282] According to another specific aspect, the present invention relates to an antigen binding domain comprising a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13 or 15, or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or 16.

[0283] According to another specific aspect, the present invention relates to an antigen binding domain comprising:

[0284] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 2;

[0285] (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 3 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 4;

[0286] (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 5 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 6;

[0287] (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 8;

[0288] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 9 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 10;

[0289] (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 11 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 12;

[0290] (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14; or

[0291] (8) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0292] According to another specific aspect, the antigen binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156 or 171-184, or having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: NO:117, 118, 121, 122, 123, 126, 127, 131, 132, 133, 134, 139, 140, 141, 145, 146, 147, 151, 152, 153, 157, 158 or 185-198 has a light chain variable region of a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical.

[0293] According to another specific aspect, the antigen binding domain is humanized and comprises:

[0294] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0295] (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0296] (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0297] (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0298] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 117;

[0299] (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0300] (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0301] (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0302] (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0303] (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0304] (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0305] (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0306] (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0307] (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0308] (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 126;

[0309] (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 127;

[0310] (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0311] (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0312] (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0313] (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0314] (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0315] (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0316] (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0317] (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0318] (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0319] (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0320] (27) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0321] (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0322] (29) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0323] (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0324] (31) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0325] (32) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0326] (33) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0327] (34) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0328] (35) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0329] (36) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0330] (37) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0331] (38) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0332] (39) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0333] (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0334] (41) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0335] (42) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0336] (43) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0337] (44) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0338] (45) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0339] (46) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0340] (47) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151;

[0341] (48) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 150 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 153;

[0342] (49) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0343] (50) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0344] (51) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0345] (52) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0346] (53) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0347] (54) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0348] (55) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 182 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190;

[0349] (56) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0350] (57) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0351] (58) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197;

[0352] (59) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198;

[0353] (60) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0354] (61) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0355] (62) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197; or

[0356] (63) A heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198.

[0357] According to another specific aspect, the antigen binding domain is a single chain variable fragment (scFv) that specifically binds FOLR1, preferably human FOLR1.

[0358] In certain embodiments, the encoded antigen binding domain is a humanized single chain variable fragment (scFv) that specifically binds to FOLR1, preferably human FOLR1. In certain embodiments, the antigen binding domain is a humanized single chain variable fragment (scFv) that specifically binds to FOLR1, preferably human FOLR1. In certain embodiments, the humanized single chain variable fragment (scFv) comprises a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to any one of SEQ ID NOs: 159-170. In certain embodiments, the humanized single chain variable fragment (scFv) comprises a polypeptide sequence having an amino acid sequence selected from the group consisting of SEQ ID NOs: 159-170.

[0359] According to another specific aspect, the chimeric antigen receptor comprises one or more antigen binding domains.

[0360] According to another specific aspect, the intracellular signaling domain comprises one or more co-stimulatory domains and one or more activation domains.

[0361] In another general aspect, the present invention relates to a polynucleotide comprising a separation of a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR comprises its antigen binding domain of the present invention. It will be appreciated by those skilled in the art that the coding sequence of the protein can be changed (e.g., replaced, deleted, inserted, etc.) without changing the amino acid sequence of the protein. Therefore, it will be appreciated by those skilled in the art that the nucleic acid sequence encoding its antigen binding domain of the present invention can be changed without changing the amino acid sequence of the protein.

[0362] In another general aspect, the present invention relates to a vector comprising a separated polynucleotide, the separated polynucleotide comprising a nucleic acid encoding a CAR, wherein the CAR comprises its antigen binding domain of the present invention. In view of the present disclosure, any vector known to those skilled in the art, such as a plasmid, a cosmid, a phage vector or a viral vector, can be used. In some embodiments, the vector is a recombinant expression vector, such as a plasmid. The vector may include any element of the conventional function of establishing an expression vector, for example, a promoter, a ribosome binding element, a terminator, an enhancer, a selection marker and an origin of replication. The promoter may be a constitutive, inducible or repressible promoter. Many expression vectors capable of delivering nucleic acids to cells are known in the art and can be used herein to produce their antigen binding domains in cells. Conventional cloning techniques or artificial gene synthesis can be used to generate recombinant expression vectors according to embodiments of the present invention.

[0363] In another general aspect, the present invention relates to a cell transduced with a vector comprising an isolated nucleic acid encoding the CAR of the present invention. The term "transduced" or "transduction" refers to the process of transferring or introducing an exogenous nucleic acid into a host cell. "Transduced" cells are cells that have been transduced with exogenous nucleic acids. The cells include primary test cells and their progeny. In certain embodiments, the cell is a CAR-T cell, preferably a human CAR-T cell, wherein the T cell is engineered to express the CAR of the present invention to treat diseases such as cancer. In certain embodiments, the cell is a CAR-NK cell, preferably a human CAR-NK cell, wherein the NK cell engineered to express the CAR of the present invention is used to treat diseases such as cancer.

[0364] In another general aspect, the present invention relates to a method of preparing CAR-T cells by transducing T cells with a vector comprising an isolated nucleic acid encoding the CAR of the present invention.

[0365] In another general aspect, the present invention relates to a method of producing a chimeric antigen receptor (CAR)-T cell of the present invention, the method comprising culturing a T cell comprising a nucleic acid encoding the CAR of the present invention under conditions that produce a CAR-T cell, and recovering the CAR-T cell.

[0366] In another general aspect, the present invention relates to a method of preparing CAR-NK cells by transducing NK cells with a vector comprising an isolated nucleic acid encoding the CAR of the present invention.

[0367] In another general aspect, the present invention relates to a method for producing a chimeric antigen receptor (CAR)-NK cell of the present invention, the method comprising culturing a NK cell comprising a nucleic acid encoding its CAR under conditions that produce a CAR-NK cell, and recovering the CAR-NK cell.

[0368] In another general aspect, the present invention relates to a method for generating an RNA engineered cell group comprising a chimeric antigen receptor (CAR) of the present invention. The method includes contacting a cell group with a polynucleotide comprising a separation of a nucleic acid encoding the CAR of the present invention, wherein the polynucleotides of the separation are RNA or synthetic RNA transcribed in vitro.

[0369] In another general aspect, the invention relates to a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to any one of SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184, or having a heavy chain variable region having a polypeptide sequence at least 95% identical to any one of SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184. NO:117, 118, 121, 122, 123, 126, 127, 131, 132, 133, 134, 139, 140, 141, 145, 146, 147, 151, 152, 153, 157, 158 or 185-198 has a light chain variable region of a polypeptide sequence that is at least 95% identical.

[0370] According to another specific aspect, the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof comprises:

[0371] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0372] (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0373] (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0374] (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0375] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 117;

[0376] (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0377] (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0378] (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0379] (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0380] (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0381] (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0382] (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0383] (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0384] (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0385] (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 126;

[0386] (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 127;

[0387] (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0388] (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0389] (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0390] (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0391] (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0392] (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0393] (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0394] (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0395] (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0396] (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0397] (27) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0398] (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0399] (29) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0400] (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0401] (31) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0402] (32) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0403] (33) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0404] (34) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0405] (35) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0406] (36) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0407] (37) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0408] (38) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0409] (39) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0410] (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0411] (41) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0412] (42) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0413] (43) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0414] (44) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0415] (45) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0416] (46) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0417] (47) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151;

[0418] (48) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 150 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 153;

[0419] (49) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0420] (50) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0421] (51) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0422] (52) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0423] (53) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0424] (54) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0425] (55) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 182 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190;

[0426] (56) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0427] (57) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0428] (58) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197;

[0429] (59) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198;

[0430] (60) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0431] (61) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0432] (62) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197; or

[0433] (63) A heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198.

[0434] According to another specific aspect, the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof is capable of binding to FOLR1, inducing effector-mediated tumor cell lysis, mediating the recruitment of conjugated drugs, and / or forming a bispecific antibody with another monoclonal antibody or antigen-binding fragment having a cancer killing effect.

[0435] In another general aspect, the invention relates to an isolated nucleic acid encoding a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the invention.

[0436] In another general aspect, the present invention relates to a vector comprising an isolated nucleic acid encoding a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the present invention.

[0437] In another general aspect, the invention relates to a host cell comprising a vector comprising an isolated nucleic acid encoding a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the invention.

[0438] In another general aspect, the present invention relates to a method of producing a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof of the present invention, the method comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions in which the monoclonal antibody or antigen-binding fragment is produced, and recovering the antibody or antigen-binding fragment from the cell or culture.

[0439] Pharmaceutical composition

[0440] In another general aspect, the present invention relates to a pharmaceutical composition comprising an isolated polynucleotide of the present invention, an isolated polypeptide of the present invention, a host cell of the present invention and / or an engineered immune cell of the present invention and a pharmaceutically acceptable carrier.

[0441] In another general aspect, the present invention relates to a pharmaceutical composition comprising the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the present invention and a pharmaceutically acceptable carrier.

[0442] As used herein, the term "pharmaceutical composition" means a product comprising an isolated polynucleotide of the present invention, an isolated polypeptide of the present invention, a host cell of the present invention, an engineered immune cell of the present invention, and / or a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment of the present invention and a pharmaceutically acceptable carrier. The polynucleotides, polypeptides, host cells, engineered immune cells and / or humanized anti-FOLR1 monoclonal antibodies or antigen-binding fragments of the present invention and compositions comprising the same can also be used to manufacture medicaments for the therapeutic applications mentioned herein.

[0443] As used herein, the term "carrier" refers to any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, oil, lipid, lipid-containing vesicle, microsphere, liposome encapsulation or other materials well known in the art for pharmaceutical preparations. It should be understood that the characteristics of the carrier, excipient or diluent will depend on the route of administration of the specific application. As used herein, the term "pharmaceutically acceptable carrier" refers to a non-toxic material that does not interfere with the effectiveness of the composition according to the present invention or the biological activity of the composition according to the present invention. According to a specific embodiment, in view of the present disclosure, any pharmaceutically acceptable carrier suitable for polynucleotides, polypeptides, host cells and / or engineered immune cell pharmaceutical compositions can be used in the present invention.

[0444] The preparation of pharmaceutically active ingredients and pharmaceutically acceptable carriers is known in the art, for example, Remington: The Science and Practice of Pharmacy (e.g., 2005, 21st edition, and any subsequent editions). Non-limiting examples of additional ingredients include: buffers, diluents, solvents, tonicity regulators, preservatives, stabilizers and chelating agents. One or more pharmaceutically acceptable carriers can be used to formulate the pharmaceutical compositions of the present invention.

[0445] In another general aspect, the present invention relates to a method for producing a pharmaceutical composition comprising a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof of the present invention, the method comprising combining the monoclonal antibody or the antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

[0446] How to use

[0447] In another general aspect, the present invention relates to a method for treating a subject's cancer in need thereof, the method comprising administering the CAR-T cells and / or CAR-NK cells of the present invention to the subject.Cancer may be, for example, selected from but not limited to lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, and non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0448] In another general aspect, the present invention relates to a method of targeting FOLR1 on the surface of cancer cells in a subject in need thereof, the method comprising administering to the subject in need thereof a pharmaceutical composition comprising a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof of the present invention.

[0449] In another general aspect, the present invention relates to a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof of the present invention. The cancer can be any liquid or solid cancer, for example, it can be selected from but not limited to lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0450] According to an embodiment of the present invention, the CAR-T cell or CAR-NK cell comprises a therapeutically effective amount of the expressed CAR of the present invention, and the pharmaceutical composition comprises a therapeutically effective amount of a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof. As used herein, the term "therapeutically effective amount" refers to the amount of an active ingredient or component that causes a desired biological or drug response in a subject. The therapeutically effective amount can be determined empirically and in a conventional manner, relevant to the purpose.

[0451] As used herein with respect to CAR, a therapeutically effective amount means the amount of a CAR molecule expressed in a transduced T cell or NK cell that modulates an immune response in a subject in need thereof. In addition, as used herein with respect to CAR, a therapeutically effective amount means the amount of a CAR molecule expressed in a transduced T cell or NK cell that leads to treatment of a disease, disorder or condition, prevention or slowing of the development of a disease, disorder or condition, or mitigation or complete alleviation of symptoms associated with a disease, disorder or condition.

[0452] As used herein for CAR-T cells or CAR-NK cells, a therapeutically effective amount means the amount of CAR-T cells or CAR-NK cells that regulate the immune response in a subject in need thereof. In addition, as used herein for CAR-T cells or CAR-NK cells, a therapeutically effective amount means the amount of CAR-T cells or CAR-NK cells that lead to the treatment of a disease, disorder or condition, the prevention or slowing of the progression of a disease, disorder or condition, or the alleviation or complete alleviation of symptoms associated with a disease, disorder or condition.

[0453] As used herein with respect to a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof, a therapeutically effective amount means an amount of a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof that modulates an immune response in a subject in need thereof. In addition, as used herein with respect to a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof, a therapeutically effective amount means an amount of a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof that results in treatment of a disease, disorder, or condition, prevention or slowing of the progression of a disease, disorder, or condition, or reduction or complete alleviation of symptoms associated with a disease, disorder, or condition.

[0454] According to a specific embodiment, the disease, disorder or condition to be treated is cancer, preferably a cancer selected from the group consisting of lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, as well as non-Hodgkin's lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0455] According to specific embodiments, a therapeutically effective amount refers to a therapeutic amount sufficient to achieve one, two, three, four or more of the following effects: (i) reduce or improve the severity of the disease, disorder or condition to be treated or the symptoms associated therewith; (ii) shorten the duration of the disease, disorder or condition to be treated or the symptoms associated therewith; (iii) prevent the progression of the disease, disorder or condition to be treated or the symptoms associated therewith; (iv) cause the regression of the disease, disorder or condition to be treated or the symptoms associated therewith; (v) prevent the development or onset of the disease, disorder or condition to be treated or the symptoms associated therewith; (v i) prevent the recurrence of the disease, disorder or condition being treated, or symptoms associated therewith; (vii) shorten the hospitalization of a subject suffering from the disease, disorder or condition being treated, or symptoms associated therewith; (viii) shorten the length of hospital stay of a subject suffering from the disease, disorder or condition being treated, or symptoms associated therewith; (ix) prolong the survival of a subject suffering from the disease, disorder or condition being treated, or symptoms associated therewith; (xi) inhibit or alleviate the disease, disorder or condition being treated, or symptoms associated therewith in a subject; and / or (xii) enhance or improve the prophylactic or therapeutic effect of another therapy.

[0456] The therapeutically effective amount or dose may vary depending on various factors, such as the disease, disorder or condition to be treated, the mode of administration, the target site, the physiological state of the subject (including, for example, age, weight, health), whether the subject is human or animal, other drugs administered, and whether the treatment is prophylactic or therapeutic. The therapeutic dose is optimally titrated to optimize safety and efficacy.

[0457] According to specific embodiments, the compositions described herein are formulated to be suitable for the intended route of administration to a subject. For example, the compositions described herein can be formulated to be suitable for intravenous administration, subcutaneous administration, or intramuscular administration.

[0458] The cells of the present invention can be administered in any convenient manner known to those skilled in the art. For example, the cells of the present invention can be administered to a subject by aerosol inhalation, injection, ingestion, infusion, implantation and / or transplantation. Compositions comprising the cells of the present invention can be administered intraarterially, subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, intrapleurally, by intravenous (iv) injection or intraperitoneally. In certain embodiments, the cells of the present invention can be administered with or without lymphocyte clearance of the subject.

[0459] Pharmaceutical compositions comprising cells of the invention expressing the CAR of the invention can be provided in sterile liquid preparations, typically aqueous solutions isotonic with cell suspensions, or optionally as emulsions, dispersions, etc., which are typically buffered to a selected pH. The composition may include a carrier suitable for the integrity and viability of the cells and for administration of the cell composition, such as water, saline, phosphate-buffered saline, etc.

[0460] Sterile injection solutions can be prepared by incorporating the cells of the present invention and various other ingredients as required into an appropriate amount of a suitable solvent. Such compositions may include a pharmaceutically acceptable carrier, diluent or excipient suitable for use with the cell composition and suitable for application to a subject such as a person, such as sterile water, normal saline, glucose, dextrose, etc. Suitable buffers for providing cell compositions are well known in the art. Any carrier, diluent or additive used is compatible with maintaining the integrity and viability of the cells of the present invention.

[0461] Cell of the present invention can be used in any physiologically acceptable carrier.The cell group comprising the cell of the present invention can comprise purified cell group.Those skilled in the art can easily determine the cell in the cell group using various well-known methods.The purity range of the cell group comprising the genetically modified cell of the present invention can be about 50% to about 55%, about 55% to about 60%, about 60% to about 65%, about 65% to about 70%, about 70% to about 75%, about 75% to about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95% or about 95% to about 100%.Those skilled in the art can easily adjust dosage, for example, the reduction of purity may need to increase dosage.

[0462] The cells of the present invention are typically administered at a dose of cells per kilogram of body weight (cells / kg) of the subject to whom the cells are administered. Typically, the cell dose is about 10 4 cells / kg body weight to about 10 10 cells / kg body weight, e.g. about 10 5 cells / kg body weight to about 10 9 cells / kg body weight, about 10 5 cells / kg body weight to about 10 8 cells / kg body weight, about 10 5 cells / kg body weight to about 10 7 cells / kg body weight or about 10 5 cells / kg body weight to about 10 6 In the case of systemic administration, higher doses are used than for local administration, where the immune cells of the invention are administered in the area of ​​a tumor and / or cancer. Exemplary dosage ranges include, but are not limited to, 1×10 4 cells / kg to 1×10 8 cells / kg, 2×10 4 cells / kg to 1×10 8 cells / kg, 3×10 4 cells / kg to 1×10 8 cells / kg, 4×10 4 cells / kg to 1×10 8 cells / kg, 5×10 4 cells / kg to 6×10 8 cells / kg, 7×10 4 cells / kg to 1×10 8 cells / kg, 8×10 4 cells / kg to 1×10 8 cells / kg, 9×10 4cells / kg to 1×10 8 cells / kg, 1×10 5 cells / kg to 1×10 8 cells / kg, 1×10 5 cells / kg to 9×10 7 cells / kg, 1×10 5 cells / kg to 8×10 7 cells / kg, 1×10 5 cells / kg to 7×10 7 cells / kg, 1×10 5 cells / kg to 6×10 7 cells / kg, 1×10 5 cells / kg to 5×10 7 cells / kg, 1×10 5 cells / kg to 4×10 7 cells / kg, 1×10 5 cells / kg to 4×10 7 cells / kg, 1×10 5 cells / kg to 3×10 7 cells / kg, 1×10 5 cells / kg to 2×10 7 cells / kg, 1×10 5 cells / kg to 1×10 7 cells / kg, 1×10 5 cells / kg to 9×10 6 cells / kg, 1×10 5 cells / kg to 8×10 6 cells / kg, 1×10 5 cells / kg to 7×10 6 cells / kg, 1×10 5 cells / kg to 6×10 6 cells / kg, 1×10 5 cells / kg to 5×10 6 cells / kg, 1×10 5 cells / kg to 4×10 6 cells / kg, 1×10 5 cells / kg to 4×10 6 cells / kg, 1×10 5 cells / kg to 3×10 6 cells / kg, 1×10 5 cells / kg to 2×10 6 cells / kg, 1×105 cells / kg to 1×10 6 cells / kg, 2×10 5 cells / kg to 9×10 7 cells / kg, 2×10 5 cells / kg to 8×10 7 cells / kg, 2×10 5 cells / kg to 7×10 7 cells / kg, 2×10 5 cells / kg to 6×10 7 cells / kg, 2×10 5 cells / kg to 5×10 7 cells / kg, 2×10 5 cells / kg to 4×10 7 cells / kg, 2×10 5 cells / kg to 4×10 7 cells / kg, 2×10 5 cells / kg to 3×10 7 cells / kg, 2×10 5 cells / kg to 2×10 7 cells / kg, 2×10 5 cells / kg to 1×10 7 cells / kg, 2×10 5 cells / kg to 9×10 6 cells / kg, 2×10 5 cells / kg to 8×10 6 cells / kg, 2×10 5 cells / kg to 7×10 6 cells / kg, 2×10 5 cells / kg to 6×10 6 cells / kg, 2×10 5 cells / kg to 5×10 6 cells / kg, 2×10 5 cells / kg to 4×10 6 cells / kg, 2×10 5 cells / kg to 4×10 6 cells / kg, 2×10 5 cells / kg to 3×10 6 cells / kg, 2×10 5 cells / kg to 2×10 6 cells / kg, 2×10 5 cells / kg to 1×10 6cells / kg, 3×10 5 cells / kg to 3×10 6 Cells / kg, etc. In addition, the dose may be adjusted to take into account whether a single dose or multiple doses are administered. The exact determination of what will be considered an effective dose can be based on individual factors for each subject.

[0463] As used herein, the terms "treat", "treating" and "treatment" are all intended to refer to the improvement or reversal of at least one measurable physical parameter associated with cancer, which physical parameter is not necessarily discernible in the subject, but can be discernible in the subject. The term "treatment" may also refer to causing regression of a disease, disorder or condition, preventing progression, or at least slowing progression. In a specific embodiment, "treatment" refers to alleviating, preventing the development or onset of one or more symptoms associated with a disease, disorder or condition such as a tumor or more preferably cancer, or shortening its duration. In a specific embodiment, "treatment" refers to preventing the recurrence of a disease, disorder or condition. In a specific embodiment, "treatment" refers to prolonging the survival of a subject suffering from the disease, disorder or condition. In a specific embodiment, "treatment" refers to eliminating a disease, disorder or condition in a subject.

[0464] According to a specific embodiment, a composition for treating cancer is provided. For cancer therapy, the provided composition can be used in combination with another treatment, which includes but is not limited to chemotherapy, anti-CD20 mAb, anti-TIM-3 mAb, anti-LAG-3 mAb, anti-EGFR mAb, anti-HER-2 mAb, anti-CD19 mAb, anti-CD33 mAb, anti-CD47 mAb, anti-CD73 mAb, anti-DLL-3 mAb, anti-apelin mAb, anti-TIP-1 mAb, anti-Claudin18.2 mAb, anti-CTLA-4 mAb, anti-PD-L1 mAb, anti-PD-1 mAb, other immuno-oncology drugs, anti-angiogenic agents, radiation therapy, antibody-drug conjugates (ADC), targeted therapy or other anti-cancer drugs.

[0465] According to a specific embodiment, a method for treating cancer in a subject in need thereof comprises administering to the subject a combination of a CAR-T cell and / or CAR-NK cell of the present invention and an agent that increases the efficacy of cells expressing the CAR molecule. Such agents include, but are not limited to, antibody fragments that bind to CD73, CD39, PD1, PD-L1, PD-L2, CTLA4, TIM3, or LAG3, or adenosine A2a receptor antagonists.

[0466] According to a specific embodiment, the method for treating cancer in a subject in need thereof comprises administering to the subject a combination of a CAR-T cell and / or CAR-NK cell of the present invention and an agent that improves one or more side effects associated with administering cells expressing CAR molecules. Such agents include, but are not limited to, steroids, TNFα inhibitors, or IL-6 inhibitors.

[0467] According to a specific embodiment, a method of treating cancer in a subject in need thereof comprises administering to the subject a combination of a CAR-T cell and / or CAR-NK cell of the present invention and an agent for treating a disease associated with FOLR1. Such agents include, but are not limited to, anti-FOLR1 monoclonal antibodies or bispecific antibodies.

[0468] As used herein, in the context of applying two or more therapies to a subject, the term "combination" refers to the use of more than one therapy. The use of the term "combination" does not limit the order in which the therapy is applied to the subject. For example, the first therapy (e.g., compositions as described herein) can be applied to the subject before the second therapy (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks before), at the same time or afterwards (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 16 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks after) is applied.

[0469] Implementation

[0470] The present invention also provides the following non-limiting embodiments.

[0471] Embodiment 1 is an isolated polynucleotide comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) an extracellular domain comprising at least one antigen binding domain that specifically binds to folate receptor 1 (FOLR1); (b) a hinge region; (c) a transmembrane region; and (d) an intracellular signaling domain.

[0472] Embodiment 2 is an isolated polynucleotide according to embodiment 1, wherein the antigen binding domain comprises a heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementary determining region 1 (LCDR1), LCDR2 and LCDR3 having the following polypeptide sequences, respectively:

[0473] (1) SEQ ID NO: 17, 18, 19, 41, 42 and 43;

[0474] (2) SEQ ID NOs: 20, 21, 22, 44, 45 and 46;

[0475] (3) SEQ ID NOs: 23, 24, 25, 47, 48 and 49;

[0476] (4) SEQ ID NO: 26, 27, 28, 50, 51 and 52;

[0477] (5) SEQ ID NO: 29, 30, 31, 53, 54 and 55;

[0478] (6) SEQ ID NOs: 32, 33, 34, 56, 57 and 58;

[0479] (7) SEQ ID NO:35, 36, 37, 59, 60 and 61; or

[0480] (8) SEQ ID NO: 38, 39, 40, 62, 63 and 64;

[0481] Wherein the antigen binding domain specifically binds to FOLR1, preferably human FOLR1.

[0482] Embodiment 3 is an isolated polynucleotide according to embodiment 1, wherein the antigen binding domain comprises a heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, a light chain complementary determining region 1 (LCDR1), LCDR2 and LCDR3 having the following polypeptide sequences, respectively:

[0483] (1) SEQ ID NO: 65, 66, 67, 89, 90 and 91;

[0484] (2) SEQ ID NOs: 68, 69, 70, 92, 93 and 94;

[0485] (3) SEQ ID NO:71, 72, 73, 95, 96 and 97;

[0486] (4) SEQ ID NO:74, 75, 76, 98, 99 and 100;

[0487] (5) SEQ ID NO: 77, 78, 79, 101, 102 and 103;

[0488] (6) SEQ ID NOs: 80, 81, 82, 104, 105 and 106;

[0489] (7) SEQ ID NO: 83, 84, 85, 107, 108 and 109; or

[0490] (8) SEQ ID NO: 86, 87, 88, 110, 111 and 112;

[0491] Wherein the antigen binding domain specifically binds to FOLR1, preferably human FOLR1.

[0492] Embodiment 4 is an isolated polynucleotide according to any one of embodiments 1 to 3, wherein the antigen binding domain comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13 or 15 or a light chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or 16.

[0493] Embodiment 5 is an isolated polynucleotide according to any one of embodiments 1 to 4, wherein the antigen binding domain comprises:

[0494] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 1 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 2;

[0495] (2) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 3 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 4;

[0496] (3) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 5 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 6;

[0497] (4) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 8;

[0498] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 9 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 10;

[0499] (6) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 11 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 12;

[0500] (7) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 13 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 14; or

[0501] (8) A heavy chain variable region having the polypeptide sequence of SEQ ID NO: 15 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16.

[0502] Embodiment 6 is an isolated polynucleotide according to any one of embodiments 1 to 4, wherein the antigen binding domain is humanized and comprises a heavy chain variable region having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184, or having a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: NO:117, 118, 121, 122, 123, 126, 127, 131, 132, 133, 134, 139, 140, 141, 145, 146, 147, 151, 152, 153, 157, 158 or 185-198 has a light chain variable region of a polypeptide sequence that is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical.

[0503] Embodiment 7 is the isolated polynucleotide of embodiment 6, wherein the antigen binding domain is humanized and comprises:

[0504] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0505] (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0506] (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0507] (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0508] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 117;

[0509] (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0510] (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0511] (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0512] (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0513] (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0514] (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0515] (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0516] (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0517] (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0518] (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 126;

[0519] (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 127;

[0520] (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0521] (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0522] (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0523] (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0524] (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0525] (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0526] (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0527] (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0528] (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0529] (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0530] (27) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0531] (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0532] (29) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0533] (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0534] (31) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0535] (32) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0536] (33) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0537] (34) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0538] (35) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0539] (36) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0540] (37) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0541] (38) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0542] (39) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0543] (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0544] (41) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0545] (42) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0546] (43) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0547] (44) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0548] (45) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0549] (46) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0550] (47) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151;

[0551] (48) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 150 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 153;

[0552] (49) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0553] (50) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0554] (51) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0555] (52) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0556] (53) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0557] (54) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0558] (55) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 182 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190;

[0559] (56) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0560] (57) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0561] (58) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197;

[0562] (59) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198;

[0563] (60) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0564] (61) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0565] (62) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197; or

[0566] (63) A heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198.

[0567] Embodiment 8 is an isolated polynucleotide according to any one of embodiments 1 to 7, wherein the antigen binding domain is a single chain variable fragment (scFv) that specifically binds FOLR1, preferably human FOLR1.

[0568] Embodiment 9 is an isolated polynucleotide according to embodiment 8, wherein the single chain variable fragment (scFv) is humanized.

[0569] Embodiment 10 is an isolated polynucleotide according to embodiment 8 or 9, wherein the single chain variable fragment (scFv) comprises a polypeptide sequence at least 95% identical to any one of SEQ ID NOs: 159-170.

[0570] Embodiment 11 is an isolated polynucleotide according to any one of embodiments 1 to 10, wherein the chimeric antigen receptor (CAR) comprises one or more antigen binding domains.

[0571] Embodiment 12 is an isolated polynucleotide according to any one of embodiments 1 to 11, wherein the intracellular signaling domain of the CAR comprises one or more co-stimulatory domains and one or more activation domains.

[0572] Embodiment 13 is a chimeric antigen receptor (CAR) encoded by the isolated polynucleotide according to any one of embodiments 1 to 12.

[0573] Embodiment 14 is a vector comprising the isolated polynucleotide according to any one of embodiments 1 to 12.

[0574] Embodiment 15 is a host cell comprising the vector according to embodiment 14.

[0575] Embodiment 16 is a host cell according to embodiment 15, wherein the cell is a CAR-T cell, preferably a human CAR-T cell.

[0576] Embodiment 17 is a host cell according to embodiment 15, wherein the cell is a CAR-NK cell, preferably a human CAR-NK cell.

[0577] Embodiment 18 is a method for preparing a host cell expressing a chimeric antigen receptor (CAR), the method comprising transducing a T cell with the vector according to embodiment 14.

[0578] Embodiment 19 is a method for producing a chimeric antigen receptor (CAR)-T cell, the method comprising culturing a T cell comprising an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) according to any one of Embodiments 1 to 12 under conditions that produce the CAR-T cell, and recovering the CAR-T cell.

[0579] Embodiment 20 is a method for preparing a host cell expressing a chimeric antigen receptor (CAR), the method comprising transducing NK cells with the vector according to embodiment 14.

[0580] Embodiment 21 is a method for producing chimeric antigen receptor (CAR)-NK cells, the method comprising culturing NK cells comprising an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) according to any one of Embodiments 1 to 12 under conditions that produce the CAR-NK cells, and recovering the CAR-NK cells.

[0581] Embodiment 22 is a method for generating a cell comprising a chimeric antigen receptor (CAR), the method comprising contacting the cell with an isolated polynucleotide comprising a nucleic acid encoding a chimeric antigen receptor (CAR) according to any one of embodiments 1 to 12, wherein the isolated polynucleotide is an in vitro transcribed RNA or a synthetic RNA.

[0582] Embodiment 23 is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject the host cell according to any one of Embodiments 15 to 17.

[0583] Embodiment 24 is a method according to embodiment 23, wherein the cancer is selected from lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, and non-Hodgkin lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0584] Embodiment 25 is a method according to embodiment 23 or 24, which further comprises administering to the subject in need thereof an agent that increases the efficacy of cells expressing CAR.

[0585] Embodiment 26 is a method according to embodiment 23 or 24, which further comprises administering to the subject in need thereof an agent that improves one or more side effects associated with the administration of cells expressing CAR.

[0586] Embodiment 27 is a method according to embodiment 23 or 24, further comprising administering to said subject in need thereof an agent for treating a disease associated with FOLR1.

[0587] Embodiment 28 is a humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to any one of SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184, or having a polypeptide sequence at least 95% identical to any one of SEQ ID NO: 113, 114, 115, 116, 119, 120, 124, 125, 128, 129, 130, 136, 137, 138, 142, 143, 144, 148, 149, 150, 154, 155, 156, or 171-184. NO:117, 118, 121, 122, 123, 126, 127, 131, 132, 133, 134, 139, 140, 141, 145, 146, 147, 151, 152, 153, 157, 158 or 185-198 has a light chain variable region of a polypeptide sequence that is at least 95% identical.

[0588] Embodiment 29 is a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to Embodiment 28, wherein the antibody or antigen-binding fragment thereof comprises:

[0589] (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0590] (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0591] (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0592] (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0593] (5) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 117;

[0594] (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0595] (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117;

[0596] (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118;

[0597] (9) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0598] (10) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0599] (11) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 119 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0600] (12) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 121;

[0601] (13) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 122;

[0602] (14) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 120 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 123;

[0603] (15) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 126;

[0604] (16) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 124 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 127;

[0605] (17) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0606] (18) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0607] (19) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0608] (20) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 128 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0609] (21) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0610] (22) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0611] (23) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0612] (24) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 129 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0613] (25) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131;

[0614] (26) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 132;

[0615] (27) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 133;

[0616] (28) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 130 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 134;

[0617] (29) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0618] (30) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0619] (31) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 136 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0620] (32) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0621] (33) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0622] (34) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 137 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0623] (35) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 139;

[0624] (36) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 140;

[0625] (37) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 138 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 141;

[0626] (38) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0627] (39) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0628] (40) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 142 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0629] (41) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0630] (42) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0631] (43) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 143 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0632] (44) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 145;

[0633] (45) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 146;

[0634] (46) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 144 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 147;

[0635] (47) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 148 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 151;

[0636] (48) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 150 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 153;

[0637] (49) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0638] (50) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0639] (51) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 171 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0640] (52) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 185;

[0641] (53) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 186;

[0642] (54) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 173 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 187;

[0643] (55) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 182 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 190;

[0644] (56) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0645] (57) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0646] (58) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197;

[0647] (59) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 198;

[0648] (60) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 195;

[0649] (61) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 196;

[0650] (62) a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 197; or

[0651] (63) A heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198.

[0652] Embodiment 30 is a humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to embodiment 28 or 29, wherein the monoclonal antibody or antigen-binding fragment thereof is capable of binding to FOLR1, inducing effector-mediated tumor cell lysis, mediating the recruitment of conjugated drugs, and / or forming a bispecific antibody with another monoclonal antibody or antigen-binding fragment having cancer killing effects.

[0653] Embodiment 31 is an isolated nucleic acid encoding the anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 28 to 30.

[0654] Embodiment 32 is a vector comprising the isolated nucleic acid according to embodiment 31.

[0655] Embodiment 33 is a host cell comprising the vector according to embodiment 32.

[0656] Embodiment 34 is a pharmaceutical composition comprising the anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to any one of Embodiments 28 to 30 and a pharmaceutically acceptable carrier.

[0657] Embodiment 35 is a method of targeting FOLR1 on the surface of cancer cells in a subject in need thereof, the method comprising administering to the subject in need thereof a pharmaceutical composition comprising the humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to any one of Embodiments 28 to 30.

[0658] Embodiment 36 is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition according to Embodiment 34.

[0659] Embodiment 37 is a method according to embodiment 36, wherein the cancer is selected from lung cancer, gastric cancer, colon cancer, hepatocellular carcinoma, renal cell carcinoma, bladder urothelial carcinoma, metastatic melanoma, breast cancer, ovarian cancer, cervical cancer, head and neck cancer, pancreatic cancer, glioma, glioblastoma and other solid tumors, and non-Hodgkin lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), multiple myeloma (MM), acute myeloid leukemia (AML) and other liquid tumors.

[0660] Embodiment 38 is a method of producing an anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to any one of embodiments 28 to 30, the method comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions in which the monoclonal antibody or antigen-binding fragment is produced, and recovering the antibody or antigen-binding fragment from the cell or culture.

[0661] Embodiment 39 is a method of producing the pharmaceutical composition according to embodiment 34, the method comprising combining the monoclonal antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

[0662] Example

[0663] Example 1: Identification of an antigen binding domain that specifically binds to FOLR1

[0664] The antigen binding domain that specifically binds to FOLR1 is an anti-FOLR1 mAb isolated and sequenced as described in PCT / US2019 / 021084, filed on March 7, 2019, which is incorporated herein by reference in its entirety.

[0665] The sequences of the heavy chain variable regions and light chain variable regions of the antigen binding domain that specifically binds to FOLR1 are provided in Tables 1 and 2, and the CDR regions of the antigen binding domain that specifically binds to FOLR1 are provided in Tables 3 to 6.

[0666] Table 1: Heavy chain variable region sequences of antigen binding domains that specifically bind to FOLR1

[0667]

[0668] VH: heavy chain variable region

[0669] Table 2: Light chain variable region sequences of antigen binding domains that specifically bind to FOLR1

[0670]

[0671] VL: light chain variable region

[0672] Table 3: Heavy chain CDR regions 1 to 3 of the antigen binding domain that specifically binds to FOLR1

[0673] Name HC CDR1 NO HC CDR2 NO HC CDR3 NO F4 GFTFSDYG 17 ISSGSNTI 18 ARLAEWDVAY 19 F5 GFTFSNYG 20 ISSGGSYT 21 STQGSSGYVGY 22 F7 GYSFTDYN 23 IDPNYGTT 24 AIKGYGNPAAY 25 F8 GFTLSTYA 26 ISGGGGDT 27 ARQSHYGSSYYFDN 28 F10 GYAFSSSW 29 IYPGDGYT 30 TRHGDFPYWYFDV 31 F17 GFTFSDFG 32 MSYTPGTF 33 ARVHVGTVDY 34 F19 GFTFSDYW 35 IGNKFHNYET 36 TKLGRGYYVMDY 37 F20 GYTFASYY 38 INPRSGGT 39 SRSGRLRGFYTMDY 40

[0674] HC: heavy chain; CDR: complementarity determining region; ID: SEQ ID NO

[0675] The HC CDRs of the antigen-binding domain that specifically binds to FOLR1 were determined using the IMGT method (Lefranc, M.-P. et al., Nucleic Acids Res., 1999, Vol. 27, pp. 209-212).

[0676] Table 4: Light chain CDR regions 1 to 3 of antigen binding domains that specifically bind to FOLR1

[0677] Name LC CDR1 NO LC CDR2 NO LC CDR3 NO F4 QNINNN 41 FAS 42 QQIYSWPQLT 43 F5 QDITNF 44 YTS 45 LQYYNLWT 46 F7 QDINKY 47 YTS 48 LQYYNLWT 49 F8 ENIDSY 50 AAT 51 QHYYTTPPT 52 F10 ENIDSY 53 AAT 54 QHHYSTPPT 55 F17 QNINNN 56 YAS 57 QQSNSWPALT 58 F19 QDITNH 59 YTS 60 QQDSQHPWT 61 F20 ENVGSY 62 GAS 63 GQTYRFLT 64

[0678] LC: light chain; CDR: complementarity determining region; NO: SEQ ID NO

[0679] The LC CDRs of the antigen-binding domain that specifically binds to FOLR1 were determined using the IMGT method (Lefranc, M.-P. et al., Nucleic Acids Res., 1999, Vol. 27, pp. 209-212).

[0680] Table 5: Heavy chain CDR regions 1 to 3 of antigen binding domains that specifically bind to FOLR1

[0681] Name HC CDR1 NO HC CDR2 NO HC CDR3 NO F4 GFTFSDYGMH 65 FISSGSNTIYYADIVKG 66 ARLAEWDVAY 67 F5 GFTFSNYGMS 68 TISSGGSYTYYPDSVKG 69 STQGSSGYVGY 70 F7 GYSFTDYNMN 71 VIDPNYGTTNYNQKFVG 72 AIKGYGNPAAY 73 F8 GFTLSTYAMS 74 TISGGGGDTYHLDTVKG 75 ARQSHYGSSYYFDN 76 F10 GYAFSSSWMN 77 RIYPGDGYTHYNGMFKG 78 TRHGDFPYWYFDV 79 F17 GFTFSDFGMH 80 YMSYTPGTFHYADTVKD 81 ARVHVGTVDY 82 F19 GFTFSDYWMN 83 QIGNKFHNYETYYSDSVKG 84 TKLGRGYYVMDY 85 F20 GYTFASYYLY 86 EINPRSGGTNFNEKFKS 87 SRSGRLRGFYTMDY 88

[0682] HC: heavy chain; CDR: complementarity determining region; NO: SEQ ID NO

[0683] The HC CDRs of the antigen binding domain that specifically binds to FOLR1 were determined using a combination of the IMGT (Lefranc, M.-P. et al., Nucleic Acids Res., 1999, Vol. 27, pp. 209-212) and Kabat (Elvin A. Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, 1991) methods.

[0684] Table 6: Light chain CDR regions 1 to 3 of antigen binding domains that specifically bind to FOLR1

[0685] Name LC CDR1 NO LC CDR2 NO LC CDR3 NO F4 RASQNINNNLH 89 FASQSIS 90 QQIYSWPQLT 91 F5 KASQDITNFIG 92 YTSILES 93 LQYYNLWT 94 F7 KASQDINKYLA 95 YTSILES 96 LQYYNLWT 97 F8 RVSENIDSYLA 98 AATNLAD 99 QHYYTTPPT 100 F10 RASENIDSYLA 101 AATNLAV 102 QHHYSTPPT 103 F17 RASQNINNNLH 104 YASQSIS 105 QQSNSWPALT 106 F19 RASQDITNHLN 107 YTSRLHS 108 QQQDSQHPWT 109 F20 KAGENVGSYVS 110 GASNRYT 111 GQTYRFLT 112

[0686] LC: light chain; CDR: complementarity determining region; NO: SEQ ID NO

[0687] The LC CDRs of the antigen binding domain that specifically binds to FOLR1 were determined using a combination of the IMGT (Lefranc, M.-P. et al., Nucleic Acids Res., 1999, Vol. 27, pp. 209-212) and Kabat (Elvin A. Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, 1991) methods.

[0688] Example 2: Humanization of mouse anti-FOLR1 mAb

[0689] The mouse anti-FOLR1 mAb was humanized to reduce the immunogenic potential when used in human patients, as described in PCT / US2019 / 021084 filed on March 7, 2019, which is incorporated herein by reference in its entirety. The sequences of the humanized VH and VL regions are shown in Table 7. The humanized VH and VL are named as follows: F5-H1 refers to the H1 sequence of the humanized VH of mouse mAb F5; F5-L1 refers to the L1 sequence of the humanized VL of mouse mAb F5. All other humanized VH and VL regions use the same naming convention.

[0690] Table 7: Heavy chain variable region and light chain variable region sequences of humanized antigen binding domains that specifically bind to FOLR1

[0691]

[0692]

[0693]

[0694] The humanized VH and VL regions were fused to the constant regions of human IgG1 heavy chain and kappa light chain, respectively. The humanized mAbs were named as follows: F5-H1L1 refers to the mAb with F5-H1 heavy chain variable region and F5-L1 light chain variable region; all other humanized mAbs used the same naming convention.

[0695] Example 3: Conversion of humanized mAb to single-chain variable fragment (scFv)

[0696] The humanized mAbs were converted into scFvs, each of which consisted of a VH and a VL with (G 4 S) n Linker (where "n" represents G 4The VH region or VL region was placed at the N-terminus of the fusion protein to identify the most effective scFv design. The sequences of the designed scFvs are shown in Table 8. The scFvs are named as follows: F5-H2 (G 4 S) 3 L2 refers to the heavy chain variable region of F5-H2, (G 4 S) 3 scFvs containing the linker and F5-L2 light chain variable region; all other scFvs follow the same naming convention.

[0697] Table 9: Sequences of humanized scFvs that specifically bind to FOLR1

[0698]

[0699]

[0700] Example 4: ELISA binding analysis of humanized mAb and scFv

[0701] To test the binding of mAbs and scFv to human FOLR1, mAbs and scFvs were produced by transient expression in ExpiCHO or Expi293 cells. 4 scFv fusion protein fused with S linker and human IgG4 Fc (scFv, G 4 The method of ELISA binding analysis of mAb or humanized scFv is described in PCT / US2019 / 021084 filed on March 7, 2019. The results of ELISA assay are shown in Figures 1A to 1L and Figures 3A to 3G middle.

[0702] Example 5: FACS analysis of mAbs and humanized scFv

[0703] The mAbs and scFv fusion proteins were also tested for their ability to bind to SK-OV-3 cells expressing endogenous FOLR1. The FACS analysis method for mAbs or humanized scFvs is described in PCT / US2019 / 021084, filed on March 7, 2019, with minor modifications. SK-OV-3 cells were plated at 100,000 cells / well. In each well of the plate, propidium iodide was incubated with the secondary antibody to label dead cells. The binding results are shown in FIG. 2A to FIG. 2E and FIG. 4A to FIG. 4G middle.

[0704] Example 6: Construction of chimeric antigen receptor constructs comprising anti-FOLR1 monoclonal antibodies or antigen-binding fragments thereof

[0705] To construct CAR, VH, VL and (G 4 Each mAb was converted into a scFv using a S)n linker, and the scFv was fused to the N-terminus of the hinge and transmembrane domains derived from human CD8α (aa 114-188, Boursier JP et al., J Biol Chem., 1993, Vol. 268, No. 3, pp. 2013 to 2020). By fusing the intracellular costimulatory domain of CD28 (aa162-202, Aruffo A and seed B, Proc Natl Acad Sci USA, 1987, Vol. 84, No. 23, pp. 8573 to 8577) followed by the activation domain from the CD3ζ chain (aa 52-162, Letournew F and Klausner RD, Proc. Nat'l. Acad. Sci. USA, 1991, Vol. 88, No. 20, pp. 8905 to 8909) to construct the C-terminal intracellular signaling domain of CAR. Using standard molecular biology cloning techniques, the DNA sequence encoding CAR is assembled and cloned into a retroviral vector to generate a CAR construct.

[0706] Those skilled in the art will appreciate that changes may be made to the above embodiments without departing from the broad inventive concept of the present invention. Therefore, it should be understood that the present invention is not limited to the specific embodiments disclosed, but the present invention is intended to cover modifications within the spirit and scope of the present invention as defined by this specification. Sequence Listing <110> Dongguan Fanenshi Biopharmaceutical Co., Ltd. <120> Humanized anti-folate receptor 1 chimeric antigen receptor and its use <130> P2021-2253 <150> US 62 / 832,975 <151> 2019-04-12 <150> US 62 / 863,330 <151> 2019-06-19 <150> US 62 / 931,988 <151> 2019-11-07 <160> 198 <170> PatentIn version 3.5 <210> 1 <211> 117 <212> PRT <213> Artificial Sequence <400> 1 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Glu Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Ser Ser Gly Ser Asn Thr Ile Tyr Tyr Ala Asp Ile Val 50 55 60 Lys Gly Arg Phe Ala Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln Met Ala Ser Leu Arg Ser Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Arg Leu Ala Glu Trp Asp Val Ala Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ala 115 <210> 2 <211> 108 <212> PRT <213> Artificial Sequence <400> 2 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Thr Pro Gly 1 5 10 15 Asp Arg Ile Ser Leu Ser Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile 35 40 45 Lys Phe Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile Asn Gly Val Glu Thr 65 70 75 80 Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ile Tyr Ser Trp Pro Gln 85 90 95 Leu Thr Phe Gly Ala Gly Thr Arg Leu Glu Leu Lys 100 105 <210> 3 <211> 118 <212> PRT <213> Artificial Sequence <400> 3 Glu Val Gln Leu Val Glu Ser Gly Gly Glu Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Thr Pro Asp Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Asp Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 4 <211> 106 <212> PRT <213> Artificial Sequence <400> 4 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Phe Leu Gly 1 5 10 15 Gly Lys Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Ile Gly Trp Tyr Gln His Lys Pro Gly Lys Gly Pro Arg Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Ile Leu Glu Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Ser Phe Ser Ile Ser Asn Leu Glu Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 5 <211> 118 <212> PRT <213> Artificial Sequence <400> 5 Glu Phe Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr 20 25 30 Asn Met Asn Trp Val Lys Gln Ser Asn Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Asn Tyr Gly Thr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Val Gly Lys Ala Thr Leu Thr Val Asp Gln Ser Ser Ile Thr Ala Tyr 65 70 75 80 Met Gln Leu Asn Ser Leu Thr Ala Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Ile Lys Gly Tyr Gly Asn Pro Ala Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala 115 <210> 6 <211> 106 <212> PRT <213> Artificial Sequence <400> 6 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Gly Lys Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Asn Lys Tyr 20 25 30 Leu Ala Trp Tyr Gln His Glu Pro Gly Lys Gly Pro Arg Leu Leu Ile 35 40 45 Arg Tyr Thr Ser Ile Leu Glu Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Ser Phe Ser Ile Ser Asn Leu Glu Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 7 <211> 121 <212> PRT <213> Artificial Sequence <400> 7 Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Gly Gly Gly Gly Asp Thr Tyr His Leu Asp Thr Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 8 <211> 107 <212> PRT <213> Artificial Sequence <400> 8 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Ile Val Thr Ile Ile Cys Arg Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Ser 65 70 75 80 Glu Asp Val Ala Arg Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Asp Ile Lys 100 105 <210> 9 <211> 120 <212> PRT <213> Artificial Sequence <400> 9 Gln Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Arg Gln Arg Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Tyr Thr His Tyr Asn Gly Met Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Gly Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val Trp Gly Thr 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 10 <211> 107 <212> PRT <213> Artificial Sequence <400> 10 Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Glu Ser Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Val Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Thr Leu Lys Ile Asn Ser Leu Gln Ser 65 70 75 80 Glu Asp Val Ala Arg Tyr Tyr Cys Gln His His Tyr Ser Thr Pro Pro 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 11 <211> 117 <212> PRT <213> Artificial Sequence <400> 11 Asp Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Arg Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe 20 25 30 Gly Met His Trp Ile Arg Gln Ala Pro Glu Arg Gly Leu Glu Trp Val 35 40 45 Ala Tyr Met Ser Tyr Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val 50 55 60 Lys Asp Arg Phe Phe Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln Met Thr Ser Leu Arg Ser Asp Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Arg Val His Val Gly Thr Val Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 <210> 12 <211> 108 <212> PRT <213> Artificial Sequence <400> 12 Asp Val Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Thr Pro Gly 1 5 10 15 Asp Ser Val Ser Leu Ser Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Phe Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Asn Val Glu Thr 65 70 75 80 Glu Asp Phe Gly Ile Tyr Phe Cys Gln Gln Ser Asn Ser Trp Pro Ala 85 90 95 Leu Thr Phe Gly Thr Gly Thr Lys Val Glu Leu Lys 100 105 <210> 13 <211> 121 <212> PRT <213> Artificial Sequence <400> 13 Glu Val Lys Leu Asp Glu Thr Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Pro Met Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ser Pro Asp Lys Gly Leu Glu Trp Val 35 40 45 Ala Gln Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ser Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Gly Ile Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 14 <211> 107 <212> PRT <213> Artificial Sequence <400> 14 Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Leu Asn Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Asp Gly Thr Phe Gln Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 15 <211> 121 <212> PRT <213> Artificial Sequence <400> 15 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Gln Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr 20 25 30 Tyr Leu Tyr Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Val Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 16 <211> 106 <212> PRT <213> Artificial Sequence(Artificial Sequence) <400> 16 Asn Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Val Ser Val Gly 1 5 10 15 Glu Arg Val Thr Leu Ser Cys Lys Ala Gly Glu Asn Val Gly Ser Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Glu Gln Ser Pro Glu Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Asp Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr 85 90 95 Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 17 <211> 8 <212> PRT <213> Artificial Sequence <400> 17 Gly Phe Thr Phe Ser Asp Tyr Gly 1 5 <210> 18 <211> 8 <212> PRT <213> Artificial Sequence <400> 18 Ile Ser Ser Gly Ser Asn Thr Ile 1 5 <210> 19 <211> 10 <212> PRT <213> Artificial Sequence <400> 19 Ala Arg Leu Ala Glu Trp Asp Val Ala Tyr 1 5 10 <210> 20 <211> 8 <212> PRT <213> Artificial Sequence <400> 20 Gly Phe Thr Phe Ser Asn Tyr Gly 1 5 <210> twenty one <211> 8 <212> PRT <213> Artificial Sequence <400> twenty one Ile Ser Ser Gly Gly Ser Tyr Thr 1 5 <210> twenty two <211> 11 <212> PRT <213> Artificial Sequence <400> twenty two Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr 1 5 10 <210> twenty three <211> 8 <212> PRT <213> Artificial Sequence <400> twenty three Gly Tyr Ser Phe Thr Asp Tyr Asn 1 5 <210> twenty four <211> 8 <212> PRT <213> Artificial Sequence <400> twenty four Ile Asp Pro Asn Tyr Gly Thr Thr 1 5 <210> 25 <211> 11 <212> PRT <213> Artificial Sequence <400> 25 Ala Ile Lys Gly Tyr Gly Asn Pro Ala Ala Tyr 1 5 10 <210> 26 <211> 8 <212> PRT <213> Artificial Sequence <400> 26 Gly Phe Thr Leu Ser Thr Tyr Ala 1 5 <210> 27 <211> 8 <212> PRT <213> Artificial Sequence <400> 27 Ile Ser Gly Gly Gly Gly Asp Thr 1 5 <210> 28 <211> 14 <212> PRT <213> Artificial Sequence <400> 28 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn 1 5 10 <210> 29 <211> 8 <212> PRT <213> Artificial Sequence <400> 29 Gly Tyr Ala Phe Ser Ser Ser Trp 1 5 <210> 30 <211> 8 <212> PRT <213> Artificial Sequence <400> 30 Ile Tyr Pro Gly Asp Gly Tyr Thr 1 5 <210> 31 <211> 13 <212> PRT <213> Artificial Sequence <400> 31 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val 1 5 10 <210> 32 <211> 8 <212> PRT <213> Artificial Sequence <400> 32 Gly Phe Thr Phe Ser Asp Phe Gly 1 5 <210> 33 <211> 8 <212> PRT <213> Artificial Sequence <400> 33 Met Ser Tyr Thr Pro Gly Thr Phe 1 5 <210> 34 <211> 10 <212> PRT <213> Artificial Sequence <400> 34 Ala Arg Val His Val Gly Thr Val Asp Tyr 1 5 10 <210> 35 <211> 8 <212> PRT <213> Artificial Sequence <400> 35 Gly Phe Thr Phe Ser Asp Tyr Trp 1 5 <210> 36 <211> 10 <212> PRT <213> Artificial Sequence <400> 36 Ile Gly Asn Lys Phe His Asn Tyr Glu Thr 1 5 10 <210> 37 <211> 12 <212> PRT <213> Artificial Sequence <400> 37 Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr 1 5 10 <210> 38 <211> 8 <212> PRT <213> Artificial Sequence <400> 38 Gly Tyr Thr Phe Ala Ser Tyr Tyr 1 5 <210> 39 <211> 8 <212> PRT <213> Artificial Sequence <400> 39 Ile Asn Pro Arg Ser Gly Gly Thr 1 5 <210> 40 <211> 14 <212> PRT <213> Artificial Sequence <400> 40 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr 1 5 10 <210> 41 <211> 6 <212> PRT <213> Artificial Sequence <400> 41 Gln Asn Ile Asn Asn Asn 1 5 <210> 42 <211> 3 <212> PRT <213> Artificial Sequence <400> 42 Phe Ala Ser 1 <210> 43 <211> 10 <212> PRT <213> Artificial Sequence <400> 43 Gln Gln Ile Tyr Ser Trp Pro Gln Leu Thr 1 5 10 <210> 44 <211> 6 <212> PRT <213> Artificial Sequence <400> 44 Gln Asp Ile Thr Asn Phe 1 5 <210> 45 <211> 3 <212> PRT <213> Artificial Sequence <400> 45 Tyr Thr Ser 1 <210> 46 <211> 8 <212> PRT <213> Artificial Sequence <400> 46 Leu Gln Tyr Tyr Asn Leu Trp Thr 1 5 <210> 47 <211> 6 <212> PRT <213> Artificial Sequence <400> 47 Gln Asp Ile Asn Lys Tyr 1 5 <210> 48 <211> 3 <212> PRT <213> Artificial Sequence <400> 48 Tyr Thr Ser 1 <210> 49 <211> 8 <212> PRT <213> Artificial Sequence <400> 49 Leu Gln Tyr Tyr Asn Leu Trp Thr 1 5 <210> 50 <211> 6 <212> PRT <213> Artificial Sequence <400> 50 Glu Asn Ile Asp Ser Tyr 1 5 <210> 51 <211> 3 <212> PRT <213> Artificial Sequence <400> 51 Ala Ala Thr 1 <210> 52 <211> 9 <212> PRT <213> Artificial Sequence <400> 52 Gln His Tyr Tyr Thr Thr Pro Pro Thr 1 5 <210> 53 <211> 6 <212> PRT <213> Artificial Sequence <400> 53 Glu Asn Ile Asp Ser Tyr 1 5 <210> 54 <211> 3 <212> PRT <213> Artificial Sequence <400> 54 Ala Ala Thr 1 <210> 55 <211> 9 <212> PRT <213> Artificial Sequence <400> 55 Gln His His Tyr Ser Thr Pro Pro Thr 1 5 <210> 56 <211> 6 <212> PRT <213> Artificial Sequence <400> 56 Gln Asn Ile Asn Asn Asn 1 5 <210> 57 <211> 3 <212> PRT <213> Artificial Sequence <400> 57 Tyr Ala Ser 1 <210> 58 <211> 10 <212> PRT <213> Artificial Sequence <400> 58 Gln Gln Ser Asn Ser Trp Pro Ala Leu Thr 1 5 10 <210> 59 <211> 6 <212> PRT <213> Artificial Sequence <400> 59 Gln Asp Ile Thr Asn His 1 5 <210> 60 <211> 3 <212> PRT <213> Artificial Sequence <400> 60 Tyr Thr Ser 1 <210> 61 <211> 9 <212> PRT <213> Artificial Sequence <400> 61 Gln Gln Asp Ser Gln His Pro Trp Thr 1 5 <210> 62 <211> 6 <212> PRT <213> Artificial Sequence <400> 62 Glu Asn Val Gly Ser Tyr 1 5 <210> 63 <211> 3 <212> PRT <213> Artificial Sequence <400> 63 Gly Ala Ser 1 <210> 64 <211> 8 <212> PRT <213> Artificial Sequence <400> 64 Gly Gln Thr Tyr Arg Phe Leu Thr 1 5 <210> 65 <211> 10 <212> PRT <213> Artificial Sequence <400> 65 Gly Phe Thr Phe Ser Asp Tyr Gly Met His 1 5 10 <210> 66 <211> 17 <212> PRT <213> Artificial Sequence <400> 66 Phe Ile Ser Ser Gly Ser Asn Thr Ile Tyr Tyr Ala Asp Ile Val Lys 1 5 10 15 Gly <210> 67 <211> 10 <212> PRT <213> Artificial Sequence <400> 67 Ala Arg Leu Ala Glu Trp Asp Val Ala Tyr 1 5 10 <210> 68 <211> 10 <212> PRT <213> Artificial Sequence <400> 68 Gly Phe Thr Phe Ser Asn Tyr Gly Met Ser 1 5 10 <210> 69 <211> 17 <212> PRT <213> Artificial Sequence <400> 69 Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val Lys 1 5 10 15 Gly <210> 70 <211> 11 <212> PRT <213> Artificial Sequence <400> 70 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr 1 5 10 <210> 71 <211> 10 <212> PRT <213> Artificial Sequence <400> 71 Gly Tyr Ser Phe Thr Asp Tyr Asn Met Asn 1 5 10 <210> 72 <211> 17 <212> PRT <213> Artificial Sequence <400> 72 Val Ile Asp Pro Asn Tyr Gly Thr Thr Asn Tyr Asn Gln Lys Phe Val 1 5 10 15 Gly <210> 73 <211> 11 <212> PRT <213> Artificial Sequence <400> 73 Ala Ile Lys Gly Tyr Gly Asn Pro Ala Ala Tyr 1 5 10 <210> 74 <211> 10 <212> PRT <213> Artificial Sequence <400> 74 Gly Phe Thr Leu Ser Thr Tyr Ala Met Ser 1 5 10 <210> 75 <211> 17 <212> PRT <213> Artificial Sequence <400> 75 Thr Ile Ser Gly Gly Gly Gly Asp Thr Tyr His Leu Asp Thr Val Lys 1 5 10 15 Gly <210> 76 <211> 14 <212> PRT <213> Artificial Sequence <400> 76 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn 1 5 10 <210> 77 <211> 10 <212> PRT <213> Artificial Sequence <400> 77 Gly Tyr Ala Phe Ser Ser Ser Trp Met Asn 1 5 10 <210> 78 <211> 17 <212> PRT <213> Artificial Sequence <400> 78 Arg Ile Tyr Pro Gly Asp Gly Tyr Thr His Tyr Asn Gly Met Phe Lys 1 5 10 15 Gly <210> 79 <211> 13 <212> PRT <213> Artificial Sequence <400> 79 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val 1 5 10 <210> 80 <211> 10 <212> PRT <213> Artificial Sequence <400> 80 Gly Phe Thr Phe Ser Asp Phe Gly Met His 1 5 10 <210> 81 <211> 17 <212> PRT <213> Artificial Sequence <400> 81 Tyr Met Ser Tyr Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val Lys 1 5 10 15 Asp <210> 82 <211> 10 <212> PRT <213> Artificial Sequence <400> 82 Ala Arg Val His Val Gly Thr Val Asp Tyr 1 5 10 <210> 83 <211> 10 <212> PRT <213> Artificial Sequence <400> 83 Gly Phe Thr Phe Ser Asp Tyr Trp Met Asn 1 5 10 <210> 84 <211> 19 <212> PRT <213> Artificial Sequence <400> 84 Gln Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ser Asp Ser 1 5 10 15 Val Lys Gly <210> 85 <211> 12 <212> PRT <213> Artificial Sequence <400> 85 Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr 1 5 10 <210> 86 <211> 10 <212> PRT <213> Artificial Sequence <400> 86 Gly Tyr Thr Phe Ala Ser Tyr Tyr Leu Tyr 1 5 10 <210> 87 <211> 17 <212> PRT <213> Artificial Sequence <400> 87 Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe Lys 1 5 10 15 Ser <210> 88 <211> 14 <212> PRT <213> Artificial Sequence <400> 88 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr 1 5 10 <210> 89 <211> 11 <212> PRT <213> Artificial Sequence <400> 89 Arg Ala Ser Gln Asn Ile Asn Asn Asn Leu His 1 5 10 <210> 90 <211> 7 <212> PRT <213> Artificial Sequence <400> 90 Phe Ala Ser Gln Ser Ile Ser 1 5 <210> 91 <211> 10 <212> PRT <213> Artificial Sequence <400> 91 Gln Gln Ile Tyr Ser Trp Pro Gln Leu Thr 1 5 10 <210> 92 <211> 11 <212> PRT <213> Artificial Sequence <400> 92 Lys Ala Ser Gln Asp Ile Thr Asn Phe Ile Gly 1 5 10 <210> 93 <211> 7 <212> PRT <213> Artificial Sequence <400> 93 Tyr Thr Ser Ile Leu Glu Ser 1 5 <210> 94 <211> 8 <212> PRT <213> Artificial Sequence <400> 94 Leu Gln Tyr Tyr Asn Leu Trp Thr 1 5 <210> 95 <211> 11 <212> PRT <213> Artificial Sequence <400> 95 Lys Ala Ser Gln Asp Ile Asn Lys Tyr Leu Ala 1 5 10 <210> 96 <211> 7 <212> PRT <213> Artificial Sequence <400> 96 Tyr Thr Ser Ile Leu Glu Ser 1 5 <210> 97 <211> 8 <212> PRT <213> Artificial Sequence <400> 97 Leu Gln Tyr Tyr Asn Leu Trp Thr 1 5 <210> 98 <211> 11 <212> PRT <213> Artificial Sequence <400> 98 Arg Val Ser Glu Asn Ile Asp Ser Tyr Leu Ala 1 5 10 <210> 99 <211> 7 <212> PRT <213> Artificial Sequence <400> 99 Ala Ala Thr Asn Leu Ala Asp 1 5 <210> 100 <211> 9 <212> PRT <213> Artificial Sequence <400> 100 Gln His Tyr Tyr Thr Thr Pro Pro Thr 1 5 <210> 101 <211> 11 <212> PRT <213> Artificial Sequence <400> 101 Arg Ala Ser Glu Asn Ile Asp Ser Tyr Leu Ala 1 5 10 <210> 102 <211> 7 <212> PRT <213> Artificial Sequence <400> 102 Ala Ala Thr Asn Leu Ala Val 1 5 <210> 103 <211> 9 <212> PRT <213> Artificial Sequence <400> 103 Gln His His Tyr Ser Thr Pro Pro Thr 1 5 <210> 104 <211> 11 <212> PRT <213> Artificial Sequence <400> 104 Arg Ala Ser Gln Asn Ile Asn Asn Asn Leu His 1 5 10 <210> 105 <211> 7 <212> PRT <213> Artificial Sequence <400> 105 Tyr Ala Ser Gln Ser Ile Ser 1 5 <210> 106 <211> 10 <212> PRT <213> Artificial Sequence <400> 106 Gln Gln Ser Asn Ser Trp Pro Ala Leu Thr 1 5 10 <210> 107 <211> 11 <212> PRT <213> Artificial Sequence <400> 107 Arg Ala Ser Gln Asp Ile Thr Asn His Leu Asn 1 5 10 <210> 108 <211> 7 <212> PRT <213> Artificial Sequence <400> 108 Tyr Thr Ser Arg Leu His Ser 1 5 <210> 109 <211> 9 <212> PRT <213> Artificial Sequence <400> 109 Gln Gln Asp Ser Gln His Pro Trp Thr 1 5 <210> 110 <211> 11 <212> PRT <213> Artificial Sequence <400> 110 Lys Ala Gly Glu Asn Val Gly Ser Tyr Val Ser 1 5 10 <210> 111 <211> 7 <212> PRT <213> Artificial Sequence <400> 111 Gly Ala Ser Asn Arg Tyr Thr 1 5 <210> 112 <211> 8 <212> PRT <213> Artificial Sequence <400> 112 Gly Gln Thr Tyr Arg Phe Leu Thr 1 5 <210> 113 <211> 118 <212> PRT <213> Artificial Sequence <400> 113 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 114 <211> 118 <212> PRT <213> Artificial Sequence <400> 114 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 115 <211> 118 <212> PRT <213> Artificial Sequence <400> 115 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 116 <211> 118 <212> PRT <213> Artificial Sequence <400> 116 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Asp Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 117 <211> 106 <212> PRT <213> Artificial Sequence <400> 117 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Ile Gly Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Ile Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 118 <211> 106 <212> PRT <213> Artificial Sequence <400> 118 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Ile Gly Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Ile Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 119 <211> 120 <212> PRT <213> Artificial Sequence <400> 119 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Tyr Thr His Tyr Asn Gly Met Phe 50 55 60 Lys Gly Arg Ala Ser Leu Thr Ala Asp Lys Ser Thr Ser Thr Gly Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Phe Phe Cys 85 90 95 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val Trp Gly Arg 100 105 110 Gly Thr Leu Val Thr Val Ser Pro 115 120 <210> 120 <211> 120 <212> PRT <213> Artificial Sequence <400> 120 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Tyr Thr His Tyr Asn Gly Met Phe 50 55 60 Lys Gly Arg Ala Ser Leu Thr Ala Asp Lys Ser Thr Ser Thr Gly Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Phe Phe Cys 85 90 95 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val Trp Gly Arg 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 121 <211> 107 <212> PRT <213> Artificial Sequence <400> 121 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 Arg Ala Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Val Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Ser Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 122 <211> 107 <212> PRT <213> Artificial Sequence <400> 122 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 Arg Ala Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Val Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Ser Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 123 <211> 107 <212> PRT <213> Artificial Sequence <400> 123 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 Arg Ala Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Val Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Ser Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 124 <211> 117 <212> PRT <213> Artificial Sequence <400> 124 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe 20 25 30 Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Tyr Met Ser Tyr Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val 50 55 60 Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val His Val Gly Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 125 <211> 117 <212> PRT <213> Artificial Sequence <400> 125 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe 20 25 30 Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Tyr Met Ser Tyr Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val 50 55 60 Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val His Val Gly Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 126 <211> 108 <212> PRT <213> Artificial Sequence <400> 126 Glu Val Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Asn Ser Trp Pro Ala 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 127 <211> 108 <212> PRT <213> Artificial Sequence <400> 127 Glu Val Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Thr 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Asn Ser Trp Pro Ala 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 128 <211> 121 <212> PRT <213> Artificial Sequence <400> 128 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr 20 25 30 Tyr Leu Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Ser Arg Ala Thr Val Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 129 <211> 121 <212> PRT <213> Artificial Sequence <400> 129 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr 20 25 30 Tyr Leu Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Ser Arg Ala Thr Val Thr Val Asp Ala Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 130 <211> 121 <212> PRT <213> Artificial Sequence <400> 130 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr 20 25 30 Tyr Leu Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Val Thr Val Asp Lys Ser Thr Asn Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 131 <211> 106 <212> PRT <213> Artificial Sequence <400> 131 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Gly Glu Asn Val Gly Ser Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 132 <211> 106 <212> PRT <213> Artificial Sequence <400> 132 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Gly Glu Asn Val Gly Ser Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 133 <211> 106 <212> PRT <213> Artificial Sequence <400> 133 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 Gly Glu Asn Val Gly Ser Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Ala Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Val Lys 100 105 <210> 134 <211> 106 <212> PRT <213> Artificial Sequence <400> 134 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 Gly Glu Asn Val Gly Ser Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Ala Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Val Lys 100 105 <210> 135 <211> 257 <212> PRT <213> Homo sapiens <400> 135 Met Ala Gln Arg Met Thr Thr Gln Leu Leu Leu Leu Leu Val Trp Val 1 5 10 15 Ala Val Val Gly Glu Ala Gln Thr Arg Ile Ala Trp Ala Arg Thr Glu 20 25 30 Leu Leu Asn Val Cys Met Asn Ala Lys His His Lys Glu Lys Pro Gly 35 40 45 Pro Glu Asp Lys Leu His Glu Gln Cys Arg Pro Trp Arg Lys Asn Ala 50 55 60 Cys Cys Ser Thr Asn Thr Ser Gln Glu Ala His Lys Asp Val Ser Tyr 65 70 75 80 Leu Tyr Arg Phe Asn Trp Asn His Cys Gly Glu Met Ala Pro Ala Cys 85 90 95 Lys Arg His Phe Ile Gln Asp Thr Cys Leu Tyr Glu Cys Ser Pro Asn 100 105 110 Leu Gly Pro Trp Ile Gln Gln Val Asp Gln Ser Trp Arg Lys Glu Arg 115 120 125 Val Leu Asn Val Pro Leu Cys Lys Glu Asp Cys Glu Gln Trp Trp Glu 130 135 140 Asp Cys Arg Thr Ser Tyr Thr Cys Lys Ser Asn Trp His Lys Gly Trp 145 150 155 160 Asn Trp Thr Ser Gly Phe Asn Lys Cys Ala Val Gly Ala Ala Cys Gln 165 170 175 Pro Phe His Phe Tyr Phe Pro Thr Pro Thr Val Leu Cys Asn Glu Ile 180 185 190 Trp Thr His Ser Tyr Lys Val Ser Asn Tyr Ser Arg Gly Ser Gly Arg 195 200 205 Cys Ile Gln Met Trp Phe Asp Pro Ala Gln Gly Asn Pro Asn Glu Glu 210 215 220 Val Ala Arg Phe Tyr Ala Ala Ala Met Ser Gly Ala Gly Pro Trp Ala 225 230 235 240 Ala Trp Pro Phe Leu Leu Ser Leu Ala Leu Met Leu Leu Trp Leu Leu 245 250 255 Ser <210> 136 <211> 117 <212> PRT <213> Artificial Sequence <400> 136 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Arg Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Ser Ser Gly Ser Asn Thr Ile Tyr Tyr Ala Asp Ile Val 50 55 60 Lys Gly Arg Phe Thr Ile Thr Arg Asn Asn Ser Thr Ser Thr Leu Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Leu Ala Glu Trp Asp Val Ala Tyr Trp Gly Ala Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 137 <211> 117 <212> PRT <213> Artificial Sequence <400> 137 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 Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Ser Ser Gly Ser Asn Thr Ile Tyr Tyr Ala Asp Ile Val 50 55 60 Lys Gly Arg Val Thr Ile Thr Arg Asn Asn Ser Thr Ser Thr Leu Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Leu Ala Glu Trp Asp Val Ala Tyr Trp Gly Ala Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 138 <211> 117 <212> PRT <213> Artificial Sequence <400> 138 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Gly Ile Ser Ser Gly Ser Asn Thr Ile Gly Tyr Ala Asp Ser Val 50 55 60 Gln Gly Arg Phe Thr Ile Ser Arg Asp Asn Gly Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Arg Leu Ala Glu Trp Asp Val Ala Tyr Trp Gly Gln Gly Thr Met 100 105 110 Val Thr Val Ser Ser 115 <210> 139 <211> 108 <212> PRT <213> Artificial Sequence <400> 139 Asp Ile Gln Leu Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Ile Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Lys Phe Ala Ser Gln Ser Ile Ser Gly Ala Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ile Tyr Ser Trp Pro Gln 85 90 95 Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 140 <211> 108 <212> PRT <213> Artificial Sequence <400> 140 Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Lys Phe Ala Ser Gln Ser Ile Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ile Tyr Ser Trp Pro Gln 85 90 95 Leu Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 141 <211> 108 <212> PRT <213> Artificial Sequence <400> 141 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Phe Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro 65 70 75 80 Glu Asp Leu Ala Val Tyr Phe Cys Gln Gln Ile Tyr Ser Trp Pro Gln 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 142 <211> 118 <212> PRT <213> Artificial Sequence <400> 142 Gln Phe 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 Asp Tyr 20 25 30 Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Trp Ile Asp Pro Asn Tyr Gly Thr Thr Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Ala Thr Leu Thr Val Asp Gln Ser Ile Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Ile Lys Gly Tyr Gly Asn Pro Ala Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 143 <211> 118 <212> PRT <213> Artificial Sequence <400> 143 Glu Phe Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr 20 25 30 Asn Phe Thr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Asn Tyr Gly Thr Thr His Tyr Ala Pro His Leu 50 55 60 Gln Gly Arg Ala Thr Leu Thr Val Asp Gln Ser Thr Ser Thr Ala Tyr 65 70 75 80 Leu Glu Leu Arg Asn Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Ile Lys Gly Tyr Gly Asn Pro Ala Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 144 <211> 118 <212> PRT <213> Artificial Sequence <400> 144 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 Asp Tyr 20 25 30 Asn Phe Thr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Asp Pro Asn Tyr Gly Thr Thr His Tyr Ala Pro His Leu 50 55 60 Gln Gly Arg Ala Thr Leu Thr Val Asp Gln Ser Thr Ser Thr Ala Tyr 65 70 75 80 Leu Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Ile Lys Gly Tyr Gly Asn Pro Ala Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 145 <211> 106 <212> PRT <213> Artificial Sequence <400> 145 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ala Ser Gln Asp Ile Asn Lys Tyr 20 25 30 Leu Ala Trp Tyr Gln His Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile 35 40 45 Arg Tyr Thr Ser Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Val Ala Val Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 146 <211> 106 <212> PRT <213> Artificial Sequence <400> 146 Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Asn Lys Tyr 20 25 30 Leu Ala Trp Tyr Gln His Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Arg Tyr Thr Ser Thr Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 147 <211> 106 <212> PRT <213> Artificial Sequence <400> 147 Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Asn Lys Tyr 20 25 30 Leu Ala Trp Tyr Gln His Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Arg Tyr Thr Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 148 <211> 121 <212> PRT <213> Artificial Sequence <400> 148 Leu Val Asn Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Phe Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 149 <211> 121 <212> PRT <213> Artificial Sequence <400> 149 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met Asn Trp Val Arg Gln Ala Pro Ala Lys Gly Leu Glu Trp Val 35 40 45 Ala Ile Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 150 <211> 121 <212> PRT <213> Artificial Sequence <400> 150 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Ala Lys Gly Leu Glu Trp Val 35 40 45 Ala Ile Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 151 <211> 107 <212> PRT <213> Artificial Sequence <400> 151 Glu Ile Gln Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 152 <211> 107 <212> PRT <213> Artificial Sequence <400> 152 Glu Ile Val Met Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys 1 5 10 15 Glu Lys Val Thr Ile Thr Cys Arg Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Gln Ser Phe Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Asp Tyr Thr Leu Thr Ile Asn Ser Leu Glu Ala 65 70 75 80 Glu Asp Ala Ala Ala Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 153 <211> 107 <212> PRT <213> Artificial Sequence <400> 153 Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys 1 5 10 15 Glu Lys Val Thr Ile Thr Cys Arg Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Gln Ser Phe Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Glu Ala 65 70 75 80 Glu Asp Ala Ala Ala Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 154 <211> 121 <212> PRT <213> Artificial Sequence <400> 154 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Glu Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Leu Arg Asp Asp Ser Lys Asn Gln 65 70 75 80 Val Ser Leu Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Ser Leu Val Thr Val Ser Ser 115 120 <210> 155 <211> 121 <212> PRT <213> Artificial Sequence <400> 155 Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Glu Tyr Asn Pro 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Leu Arg Asp Asp Ser Lys Asn Gln 65 70 75 80 Val Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Ser Leu Val Thr Val Ser Ser 115 120 <210> 156 <211> 121 <212> PRT <213> Artificial Sequence <400> 156 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ile Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 157 <211> 107 <212> PRT <213> Artificial Sequence <400> 157 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Phe Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Asn Leu Gln Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 158 <211> 107 <212> PRT <213> Artificial Sequence <400> 158 Glu Ile Val Met Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys 1 5 10 15 Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Asp Gln Ser Phe Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Gln Ser Phe Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Asn Ser Leu Glu Ala 65 70 75 80 Glu Asp Ala Ala Ala Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 159 <211> 239 <212> PRT <213> Artificial Sequence <400> 159 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 115 120 125 Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val 130 135 140 Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln 145 150 155 160 Asp Ile Thr Asn Phe Ile Gly Trp Tyr Gln His Lys Pro Gly Lys Ala 165 170 175 Pro Lys Leu Leu Ile Ser Tyr Thr Ser Ile Leu Glu Ser Gly Val Pro 180 185 190 Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile 195 200 205 Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr 210 215 220 Tyr Asn Leu Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 225 230 235 <210> 160 <211> 244 <212> PRT <213> Artificial Sequence <400> 160 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 115 120 125 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln 130 135 140 Ser Pro Ser Ser Val Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 145 150 155 160 Cys Lys Ala Ser Gln Asp Ile Thr Asn Phe Ile Gly Trp Tyr Gln His 165 170 175 Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Ser Tyr Thr Ser Ile Leu 180 185 190 Glu Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 195 200 205 Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 210 215 220 Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr Phe Gly Gly Gly Thr Lys 225 230 235 240 Val Glu Ile Lys <210> 161 <211> 239 <212> PRT <213> Artificial Sequence <400> 161 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Ile Gly Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Ile Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly 100 105 110 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser 115 120 125 Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala 130 135 140 Val Ser Gly Phe Thr Phe Ser Asn Tyr Gly Met Ser Trp Val Arg Gln 145 150 155 160 Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Thr Ile Ser Ser Gly Gly 165 170 175 Ser Tyr Thr Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser 180 185 190 Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg 195 200 205 Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ser Thr Gln Gly Ser Ser Gly 210 215 220 Tyr Val Gly Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 225 230 235 <210> 162 <211> 242 <212> PRT <213> Artificial Sequence <400> 162 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Tyr Thr His Tyr Asn Gly Met Phe 50 55 60 Lys Gly Arg Ala Ser Leu Thr Ala Asp Lys Ser Thr Ser Thr Gly Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Phe Phe Cys 85 90 95 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val Trp Gly Arg 100 105 110 Gly Thr Leu Val Thr Val Ser Pro Gly Gly Gly Gly Ser Gly Gly Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Ser 130 135 140 Thr Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala 145 150 155 160 Ser Glu Asn Ile Asp Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly 165 170 175 Arg Ala Pro Lys Leu Leu Val Tyr Ala Ala Thr Asn Leu Ala Val Gly 180 185 190 Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Tyr Thr Leu 195 200 205 Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln 210 215 220 His His Tyr Ser Thr Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu 225 230 235 240 Ile Lys <210> 163 <211> 247 <212> PRT <213> Artificial Sequence <400> 163 Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Arg Ile Tyr Pro Gly Asp Gly Tyr Thr His Tyr Asn Gly Met Phe 50 55 60 Lys Gly Arg Ala Ser Leu Thr Ala Asp Lys Ser Thr Ser Thr Gly Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Phe Phe Cys 85 90 95 Thr Arg His Gly Asp Phe Pro Tyr Trp Tyr Phe Asp Val Trp Gly Arg 100 105 110 Gly Thr Leu Val Thr Val Ser Pro Gly Gly Gly Gly Ser Gly Gly Gly 115 120 125 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met 130 135 140 Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly Asp Arg Val Thr 145 150 155 160 Ile Thr Cys Arg Ala Ser Glu Asn Ile Asp Ser Tyr Leu Ala Trp Tyr 165 170 175 Gln Gln Lys Pro Gly Arg Ala Pro Lys Leu Leu Val Tyr Ala Ala Thr 180 185 190 Asn Leu Ala Val Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly 195 200 205 Thr Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala 210 215 220 Thr Tyr Tyr Cys Gln His His Tyr Ser Thr Pro Pro Thr Phe Gly Gln 225 230 235 240 Gly Thr Lys Leu Glu Ile Lys 245 <210> 164 <211> 242 <212> PRT <213> Artificial Sequence <400> 164 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 Arg Ala Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Val Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His His Tyr Ser Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser 100 105 110 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Gln 115 120 125 Ser Gly Ala Glu Val Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys 130 135 140 Lys Ala Ser Gly Tyr Ala Phe Ser Ser Ser Trp Met Asn Trp Val Arg 145 150 155 160 Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Arg Ile Tyr Pro Gly 165 170 175 Asp Gly Tyr Thr His Tyr Asn Gly Met Phe Lys Gly Arg Ala Ser Leu 180 185 190 Thr Ala Asp Lys Ser Thr Ser Thr Gly Tyr Met Glu Leu Ser Ser Leu 195 200 205 Arg Ser Glu Asp Thr Ala Val Phe Phe Cys Thr Arg His Gly Asp Phe 210 215 220 Pro Tyr Trp Tyr Phe Asp Val Trp Gly Arg Gly Thr Leu Val Thr Val 225 230 235 240 Ser Pro <210> 165 <211> 240 <212> PRT <213> Artificial Sequence <400> 165 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe 20 25 30 Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Tyr Met Ser Tyr Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val 50 55 60 Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val His Val Gly Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Glu Val Val Leu Thr Gln Ser Pro Ala Thr Leu Ser 130 135 140 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn 145 150 155 160 Ile Asn Asn Asn Leu His Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro 165 170 175 Arg Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala 180 185 190 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 195 200 205 Ser Leu Glu Thr Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Asn 210 215 220 Ser Trp Pro Ala Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 225 230 235 240 <210> 166 <211> 245 <212> PRT <213> Artificial Sequence <400> 166 Glu Val Gln Leu Val Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe 20 25 30 Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Tyr Met Ser Tyr Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val 50 55 60 Lys Asp Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val His Val Gly Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Val Leu Thr Gln Ser 130 135 140 Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys 145 150 155 160 Arg Ala Ser Gln Asn Ile Asn Asn Asn Leu His Trp Phe Gln Gln Lys 165 170 175 Pro Gly Gln Ala Pro Arg Leu Leu Ile Lys Tyr Ala Ser Gln Ser Ile 180 185 190 Ser Gly Ile Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 195 200 205 Thr Leu Thr Ile Ser Ser Leu Glu Thr Glu Asp Phe Ala Val Tyr Phe 210 215 220 Cys Gln Gln Ser Asn Ser Trp Pro Ala Leu Thr Phe Gly Gln Gly Thr 225 230 235 240 Lys Val Glu Ile Lys 245 <210> 167 <211> 240 <212> PRT <213> Artificial Sequence <400> 167 Glu Val Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Ile Asn Asn Asn 20 25 30 Leu His Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Thr 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Ser Asn Ser Trp Pro Ala 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Gly Gly Gly Gly 100 105 110 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val 115 120 125 Glu Thr Gly Gly Gly Leu Ile Gln Pro Gly Gly Ser Leu Arg Leu Ser 130 135 140 Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Phe Gly Met His Trp Ile 145 150 155 160 Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Tyr Met Ser Tyr 165 170 175 Thr Pro Gly Thr Phe His Tyr Ala Asp Thr Val Lys Asp Arg Phe Thr 180 185 190 Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser 195 200 205 Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val His Val 210 215 220 Gly Thr Val Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 225 230 235 240 <210> 168 <211> 242 <212> PRT <213> Artificial Sequence <400> 168 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr 20 25 30 Tyr Leu Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Ser Arg Ala Thr Val Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro 130 135 140 Ser Thr Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys 145 150 155 160 Ala Gly Glu Asn Val Gly Ser Tyr Val Ser Trp Tyr Gln Gln Lys Pro 165 170 175 Gly Lys Ala Pro Lys Leu Leu Ile Tyr Gly Ala Ser Asn Arg Tyr Thr 180 185 190 Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Ala Thr Glu Phe Thr 195 200 205 Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys 210 215 220 Gly Gln Thr Tyr Arg Phe Leu Thr Phe Gly Gln Gly Thr Lys Val Glu 225 230 235 240 Val Lys <210> 169 <211> 247 <212> PRT <213> Artificial Sequence <400> 169 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Tyr 20 25 30 Tyr Leu Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Arg Ser Gly Gly Thr Asn Phe Asn Glu Lys Phe 50 55 60 Lys Ser Arg Ala Thr Val Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Ser Gly Arg Leu Arg Gly Phe Tyr Thr Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln 130 135 140 Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly Asp Arg Val 145 150 155 160 Thr Ile Thr Cys Lys Ala Gly Glu Asn Val Gly Ser Tyr Val Ser Trp 165 170 175 Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Gly Ala 180 185 190 Ser Asn Arg Tyr Thr Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser 195 200 205 Ala Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe 210 215 220 Ala Thr Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr Phe Gly Gln 225 230 235 240 Gly Thr Lys Val Glu Val Lys 245 <210> 170 <211> 242 <212> PRT <213> Artificial Sequence <400> 170 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 Gly Glu Asn Val Gly Ser Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Ala Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gly Gln Thr Tyr Arg Phe Leu Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Val Lys Gly Gly Gly Gly Ser Gly 100 105 110 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser 115 120 125 Gly Ala Glu Val Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys 130 135 140 Ala Ser Gly Tyr Thr Phe Ala Ser Tyr Tyr Leu Tyr Trp Val Arg Gln 145 150 155 160 Ala Pro Gly Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro Arg Ser 165 170 175 Gly Gly Thr Asn Phe Asn Glu Lys Phe Lys Ser Arg Ala Thr Val Thr 180 185 190 Val Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg 195 200 205 Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ser Arg Ser Gly Arg Leu Arg 210 215 220 Gly Phe Tyr Thr Met Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 225 230 235 240 Ser Ser <210> 171 <211> 121 <212> PRT <213> Artificial Sequence <400> 171 Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 172 <211> 121 <212> PRT <213> Artificial Sequence <400> 172 Gln Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Ser Gly Gly Gly Gly Asp Thr Tyr His Leu Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Met Val Thr Val Ser Ser 115 120 <210> 173 <211> 121 <212> PRT <213> Artificial Sequence <400> 173 Glu Val Gln Leu Val Val Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Ser Thr Tyr 20 25 30 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Val Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gln Ser His Tyr Gly Ser Ser Tyr Tyr Phe Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 174 <211> 121 <212> PRT <213> Artificial Sequence <400> 174 Glu Val Val Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala His Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Asp Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 175 <211> 121 <212> PRT <213> Artificial Sequence <400> 175 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala His Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Asp Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 176 <211> 121 <212> PRT <213> Artificial Sequence <400> 176 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 177 <211> 121 <212> PRT <213> Artificial Sequence <400> 177 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala His Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Asp Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 178 <211> 121 <212> PRT <213> Artificial Sequence <400> 178 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Thr Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Lys Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 179 <211> 121 <212> PRT <213> Artificial Sequence <400> 179 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Thr Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Lys Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 180 <211> 121 <212> PRT <213> Artificial Sequence <400> 180 Gln Val Val Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ser Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Phe Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 181 <211> 121 <212> PRT <213> Artificial Sequence <400> 181 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ala Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 65 70 75 80 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 182 <211> 121 <212> PRT <213> Artificial Sequence <400> 182 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Gln Ile Gly Asn Lys Phe His Asn Tyr Glu Thr Tyr Tyr Ser Asp 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Lys Asp Asp Ser Lys Asn Thr 65 70 75 80 Val Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 85 90 95 Tyr Cys Thr Lys Leu Gly Arg Gly Tyr Tyr Val Met Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 183 <211> 118 <212> PRT <213> Artificial Sequence <400> 183 Gln Val Glu Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Asp Cys Lys Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 184 <211> 118 <212> PRT <213> Artificial Sequence <400> 184 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Lys Val Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Ser Gly Gly Ser Tyr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Thr Gln Gly Ser Ser Gly Tyr Val Gly Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 185 <211> 107 <212> PRT <213> Artificial Sequence <400> 185 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 Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Asp Gly Thr Lys Val Glu Ile Lys 100 105 <210> 186 <211> 107 <212> PRT <213> Artificial Sequence <400> 186 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 Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Asn Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Asp Gly Thr Lys Val Glu Ile Lys 100 105 <210> 187 <211> 107 <212> PRT <213> Artificial Sequence <400> 187 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 Arg Val Ser Glu Asn Ile Asp Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Val 35 40 45 Tyr Ala Ala Thr Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Asp Ile Lys 100 105 <210> 188 <211> 107 <212> PRT <213> Artificial Sequence <400> 188 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Phe Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Arg Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 189 <211> 107 <212> PRT <213> Artificial Sequence <400> 189 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Phe Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Glu Ile Lys 100 105 <210> 190 <211> 107 <212> PRT <213> Artificial Sequence <400> 190 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Phe Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 191 <211> 107 <212> PRT <213> Artificial Sequence <400> 191 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ala Phe Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 192 <211> 107 <212> PRT <213> Artificial Sequence <400> 192 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 193 <211> 107 <212> PRT <213> Artificial Sequence <400> 193 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala Phe Arg Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 194 <211> 107 <212> PRT <213> Artificial Sequence <400> 194 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Thr Asn His 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Asp Ser Gln His Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 195 <211> 106 <212> PRT <213> Artificial Sequence <400> 195 Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Leu Gly Trp Tyr Gln His Lys Pro Gly Glu Ala Pro Lys Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Val Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 196 <211> 106 <212> PRT <213> Artificial Sequence <400> 196 Glu Ile Val Met Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys 1 5 10 15 Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Ile Gly Trp Tyr Gln His Lys Pro Asp Gln Ser Pro Lys Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Gln Ser Phe Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Asn Ser Leu Glu Ala 65 70 75 80 Glu Asp Ala Ala Ala Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 197 <211> 106 <212> PRT <213> Artificial Sequence <400> 197 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Leu Gly Trp Tyr Gln His Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Arg Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 198 <211> 106 <212> PRT <213> Artificial Sequence <400> 198 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asp Ile Thr Asn Phe 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Tyr Thr Ser Ile Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Leu Gln Tyr Tyr Asn Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105

Claims

1. A single-chain variable fragment (scFv) that specifically binds to FOLR1, comprising the heavy chain complementary determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementary determining region 1 (LCDR1), LCDR2 and LCDR3 of the following polypeptide sequence: SEQ ID NOs: 20, 21, 22, 44, 45 and 46, wherein the heavy chain CDRs and light chain CDRs of the antigen binding domain are determined using the IMGT method; or SEQ ID NOs: 68, 69, 70, 92, 93 and 94, wherein the heavy chain CDRs and light chain CDRs of the antigen binding domain are determined using a combination of IMGT and Kabat methods.

2. The single-chain variable fragment according to claim 1, wherein the single-chain variable fragment specifically binds to human FOLR1.

3. The single-chain variable fragment of claim 1, wherein the single-chain variable fragment comprises a heavy chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 3, 113, 114, 115, 116, 183 or 184, or a light chain variable region having a polypeptide sequence at least 95% identical to SEQ ID NO: 4, 117, 118, 195, 196, 197 or 198.

4. The single-chain variable fragment according to claim 1, wherein the single-chain variable fragment comprises: (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 3 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 4; (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (5) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (9) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (10) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 195; (11) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 196; (12) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 197; (13) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198; (14) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 195; (15) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 196; (16) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 197; or (17) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO:

198.

5. The single chain variable fragment of claim 1, wherein the single chain variable fragment (scFv) is humanized.

6. The single-chain variable fragment of claim 1, wherein the single-chain variable fragment (scFv) comprises a polypeptide sequence that is at least 95% identical to any one of SEQ ID NO: 159, 160 or 161.

7. An isolated polynucleotide encoding the single-chain variable fragment according to any one of claims 1 to 6.

8. A vector comprising the isolated polynucleotide according to claim 7.

9. A host cell comprising the vector according to claim 8.

10. A humanized anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region of a polypeptide sequence as shown in any one of SEQ ID NOs: 113, 114, 115, 116, 183 or 184, and a light chain variable region of a polypeptide sequence as shown in any one of SEQ ID NOs: 117, 118, 195, 196, 197 or 198.

11. The humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to claim 10, wherein the antibody or antigen-binding fragment thereof comprises: (1) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (2) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 113 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (3) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (4) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 114 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (5) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (6) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 115 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (7) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 117; (8) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 116 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 118; (9) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 195; (10) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 196; (11) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 197; (12) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 183 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 198; (13) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 195; (14) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 196; (15) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO: 197; or (16) a heavy chain variable region having a polypeptide sequence of SEQ ID NO: 184 and a light chain variable region having a polypeptide sequence of SEQ ID NO:

198.

12. The humanized anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to claim 10 or 11, wherein the monoclonal antibody or antigen-binding fragment thereof is capable of binding to FOLR1, inducing effector-mediated tumor cell lysis, mediating the recruitment of conjugated drugs, and / or forming a bispecific antibody with another monoclonal antibody or antigen-binding fragment having cancer killing effects.

13. An isolated nucleic acid encoding the anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to any one of claims 10-12.

14. A vector comprising the isolated nucleic acid according to claim 13.

15. A host cell comprising the vector according to claim 14. 16 . A pharmaceutical composition comprising the anti-FOLR1 monoclonal antibody or antigen-binding fragment thereof according to any one of claims 10 to 12 and a pharmaceutically acceptable carrier.

17. Use of the pharmaceutical composition according to claim 16 in the preparation of a drug for treating cancer; Wherein the cancer is selected from lung cancer, breast cancer and ovarian cancer.

18. A method for producing an anti-FOLR1 monoclonal antibody or an antigen-binding fragment thereof according to any one of claims 10 to 12, the method comprising culturing a cell comprising a nucleic acid encoding the monoclonal antibody or antigen-binding fragment under conditions in which the monoclonal antibody or antigen-binding fragment is produced, and recovering the antibody or antigen-binding fragment from the cell or culture.

19. A method of producing the pharmaceutical composition according to claim 16, comprising combining the monoclonal antibody or antigen-binding fragment thereof with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.

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