RU2777662C2 - Conjugates of antibody to ccr7 and drug - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: antibodies to CCR7, their antigen-binding fragments, and antibody-drug conjugates based on the specified antibodies or antigen-binding fragments are proposed. The invention also relates to pharmaceutical compositions and methods for the treatment or prevention of cancer, using the specified antibodies, antigen-binding fragments, and conjugates. In addition, nucleic acids, expression vectors, and host cells for the production of antibodies are disclosed.

EFFECT: invention can be used in cancer therapy in the absence of significant depletion of normal hemopoietic cells expressing CCR7.

40 cl, 22 dwg, 27 tbl, 21 ex

Description

Cross-reference to related applications

[0001] This application claims benefit from U.S. Provisional Application No. 62/454,476, filed Feb. 3, 2017, the contents of which are hereby incorporated by reference in its entirety.

SEQUENCE LIST

[0002] This application contains a sequence listing that has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. The specified ASCII copy, created on January 10, 2018, is named PAT057594-WO-PCT_SL.txt and is 387054 bytes in size.

The technical field to which the invention belongs

[0003] The present invention relates to anti-CCR7 antibodies, antibody fragments and immunoconjugates thereof, as well as their uses for the treatment or prevention of cancer.

Background of the invention

[0004] CC-chemokine receptor 7 (CCR7) was first identified in 1993 as a lymphocyte-specific receptor (see, for example, Birkenbach et al., J Virol. 1993 Apr;67(4):2209-20). Its expression is limited to immune cell subpopulations such as naive T cells, central memory T cells (Tcm), regulatory T cells (Treg), naive B cells, NK cells, and mature antigen presenting dendritic cells (DC). CCR7 regulates immune cell homing to and migration within lymphoid organs and thus plays a key role in the balance of immunity and tolerance (see, for example, Förster et al., Nat Rev Immunol. 2008 May;8(5):362 -71).

[0005] CCR7 is a class A rhodopsin-like G protein coupled receptor (GPCR) with two ligands, CCL21 and CCL19. The structure of CCR7 is not yet fully elucidated, however, several motifs required for receptor activity have been discovered (see, for example, Legler et al., Int J Biochem Cell Biol. 2014 Jul 1).

CCR7 and cancer

[0006] CCR7 (also referred to as EBI1, BLR2, CC-CKR-7, CMKBR7, CD197, and CDw197) is also known to be overexpressed on a variety of cancers, including but not limited to B-cell cancers (e.g., CLL, MCL , Burkitt's lymphoma), T-cell malignancies (eg, ATLL), HNSCC, ESCC, gastric carcinoma, NSCLC, colorectal carcinoma, pancreatic cancer, thyroid cancer, breast cancer, and cervical cancer. For example, overexpression of CCR7 on colorectal carcinoma, ESCC, pancreatic cancer, HNSCC, and gastric cancer cells has been associated with late tumor stage, lymph node metastasis, and poor prognosis for survival (see, e.g., Malietzis et al., Journal of Surgical Oncology 2015;112:86-92; Irino et al., BMC Cancer 2014, 14:291; Guo et al., Oncology Letters 5: 1572-1578, 2013; Xia et al., Oral Dis. 2015 Jan;21( 1):123-31; Du et al., Gastric Cancer. 2016 Mar 16).

[0007] In addition, for example, CCR7 expression on HNSCC cells has been shown to play a role in resistance to chemotherapy (see, for example, Wang et al., JNCI J Natl Cancer Inst (2008) 100 (7): 502-512 .). In some types of cancer, such as pancreatic cancer and nasopharyngeal carcinoma (NPC), CCR7 is known to stimulate cancer stem cell metastasis and sphere formation (see, e.g., Zhang et al., PLOS ONE 11 (8); Lun et al . ., PLOS ONE 7(12)). The role of CCR7 in cell migration, invasiveness, and EMT (epithelial-mesenchymal transition) has been described in various types of cancer such as breast and pancreatic cancer in vitro and in vivo (see, for example, Pang et al., Oncogene (2015), 1-13); Sperveslage et al., Int. J. Cancer: 131, E371-E381 (2012)). Key pathways that have been described as required for CCR7 signaling include the b-arrestin-mediated p38/ERK1/2 signaling pathway and the Rho signaling pathway (see e.g. Noor et al., J Neuroinflammation 2012 Apr 25; 9:77).

[0008] Numerous cancer-associated processes are known to induce CCR7 expression. In HNSCC, CCR7 expression has been shown to be induced by transcription factors NF-kB and AP1 by direct binding to sites in the CCR7 promoter (Mburu et al., J. Biol. Chem. 2012, 287:3581-3590). In particular, CCR7 expression is regulated by various factors in the tumor microenvironment. In this context, it is known that CCR7 expression is induced by the b-defensin 3/NF-kB pathway in HNSCC (see e.g. Mburu et al., Carcinogenesis vol.32 no.2 pp.168-174, 2010) and endothelin receptor A and hypoxia inducible factor-1 in breast tumor cells (see, for example, Wilson et al., Cancer Res 2006;66:11802-11807).

Antibody drug conjugates

[0009] Antibody drug conjugates ("ADCs") have found use in the local delivery of cytotoxic agents in the treatment of cancer (see, for example, Lambert, Curr. Opinion In Pharmacology 5:543-549, 2005). ADCs allow targeted delivery of a drug moiety that can achieve maximum efficacy with minimum toxicity. ADCs include an antibody selected for its ability to bind to the cell targeted by the therapeutic intervention, coupled to a drug selected for its cytostatic or cytotoxic activity. Thus, when the antibody binds to the target cell, the drug is delivered to the site where its therapeutic effect is required.

[0010] A variety of antibodies have been disclosed for use in ADCs that recognize and selectively bind to target cells, such as cancer cells. Despite extensive work on ADC, binding of an antibody to a particular target of interest is not sufficient to predict success in ADC applications. Examples of factors that can affect the therapeutic efficacy of ADCs (other than the inherent characteristics of the target) include various aspects that need to be modified by fine tuning, such as optimal antibody affinity as a balance between target-mediated distribution (TMDD) and performance-driven exposure, evaluation Fc-mediated functions (antibody-dependent cell-mediated cytotoxicity, ADCC), method of conjugation (site-specific or not), drug/payload ratio of such a conjugate with each antibody ("DAR" or "drug-to-antibody ratio"), cleavage or linker stability, ADC stability, and ADC tendency to aggregate.

[0011] There remains a need for antibodies, attachment methods, and cytotoxic payloads with improved properties for use as effective ADC-based therapeutic compositions and methods associated therewith.

Brief description of the present invention

[0012] Disclosed herein is an antibody or antigen-binding fragment thereof that binds to human CCR7 protein, wherein the antibody or antigen-binding fragment thereof has reduced or negligible effector function compared to a wild-type antibody of the same isotype. In one embodiment, the antibody, or antigen-binding fragment thereof, has a reduced or negligible level of antibody-dependent cell-mediated cytotoxicity (ADCC) activity. In one embodiment, the antibody, or antigen-binding fragment thereof, comprises a silent Fc region. In some embodiments, the antibody contains in the Fc region a mutation selected from: D265A; P329A; P329G; N297A; D265A and P329A; D265A and N297A; L234 and L235A; P329A, L234A and L235A; as well as P329G, L234A and L235A. In one embodiment, the antibody or antigen-binding fragment has little cell cytolysis activity. In one embodiment, the antibody, or antigen-binding fragment thereof, binds with greater affinity to cells expressing higher levels of CCR7 than to cells expressing lower levels of CCR7. In some embodiments, the antibody, or antigen-binding fragment thereof, binds with greater affinity to cancer cells that express higher levels of CCR7 than to normal cells that express lower levels of CCR7. In some embodiments, the antibody or antigen-binding fragment thereof slightly depletes normal hematopoietic cells that express CCR7.

[0013] In one embodiment, the present application discloses an antibody or antigen-binding fragment that binds CCR7, comprising:

a. a heavy chain variable region that contains HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO:1, HCDR2 (heavy chain complementarity determining region 2) of SEQ ID NO:2, and HCDR3 (heavy chain complementarity determining region 3) of SEQ ID NO:3; and a light chain variable region which contains LCDR1 (light chain complementarity determining region 1) of SEQ ID NO:17, LCDR2 (light chain complementarity determining region 2) of SEQ ID NO:18, and LCDR3 (light chain complementarity determining region 3) of SEQ ID NO:19;

b. a heavy chain variable region that contains HCDR1 of SEQ ID NO:4, HCDR2 of SEQ ID NO:5, and HCDR3 of SEQ ID NO:6; and a light chain variable region that contains LCDR1 of SEQ ID NO:20, LCDR2 of SEQ ID NO:21, and LCDR3 of SEQ ID NO:22;

c. a heavy chain variable region that contains HCDR1 of SEQ ID NO:7, HCDR2 of SEQ ID NO:8, and HCDR3 of SEQ ID NO:9; and a light chain variable region that contains LCDR1 of SEQ ID NO:23, LCDR2 of SEQ ID NO:24, and LCDR3 of SEQ ID NO:25;

d. a heavy chain variable region that contains HCDR1 of SEQ ID NO:10, HCDR2 of SEQ ID NO:11, and HCDR3 of SEQ ID NO:12; and a light chain variable region that contains LCDR1 of SEQ ID NO:26, LCDR2 of SEQ ID NO:27, and LCDR3 of SEQ ID NO:28;

e. a heavy chain variable region that contains HCDR1 of SEQ ID NO:33, HCDR2 of SEQ ID NO:34, and HCDR3 of SEQ ID NO:35; and a light chain variable region that contains LCDR1 of SEQ ID NO:49, LCDR2 of SEQ ID NO:50, and LCDR3 of SEQ ID NO:51;

f. a heavy chain variable region that contains HCDR1 of SEQ ID NO:36, HCDR2 of SEQ ID NO:37, and HCDR3 of SEQ ID NO:38; and a light chain variable region that contains LCDR1 of SEQ ID NO:52, LCDR2 of SEQ ID NO:53, and LCDR3 of SEQ ID NO:54;

g. a heavy chain variable region that contains HCDR1 of SEQ ID NO:39, HCDR2 of SEQ ID NO:40, and HCDR3 of SEQ ID NO:41; and a light chain variable region that contains LCDR1 of SEQ ID NO:55, LCDR2 of SEQ ID NO:56, and LCDR3 of SEQ ID NO:57;

h. a heavy chain variable region that contains HCDR1 of SEQ ID NO:42, HCDR2 of SEQ ID NO:43, and HCDR3 of SEQ ID NO:44; and a light chain variable region that contains LCDR1 of SEQ ID NO:58, LCDR2 of SEQ ID NO:59, and LCDR3 of SEQ ID NO:60;

i. a heavy chain variable region that contains HCDR1 of SEQ ID NO:65, HCDR2 of SEQ ID NO:66, and HCDR3 of SEQ ID NO:67; and a light chain variable region that contains LCDR1 of SEQ ID NO:81, LCDR2 of SEQ ID NO:82, and LCDR3 of SEQ ID NO:83;

j. a heavy chain variable region that contains HCDR1 of SEQ ID NO:68, HCDR2 of SEQ ID NO:69, and HCDR3 of SEQ ID NO:70; and a light chain variable region that contains LCDR1 of SEQ ID NO:84, LCDR2 of SEQ ID NO:85, and LCDR3 of SEQ ID NO:86;

k. a heavy chain variable region that contains HCDR1 of SEQ ID NO:71, HCDR2 of SEQ ID NO:72, and HCDR3 of SEQ ID NO:73; and a light chain variable region that contains LCDR1 of SEQ ID NO:87, LCDR2 of SEQ ID NO:88, and LCDR3 of SEQ ID NO:89;

l. a heavy chain variable region that contains HCDR1 of SEQ ID NO:74, HCDR2 of SEQ ID NO:75, and HCDR3 of SEQ ID NO:76; and a light chain variable region that contains LCDR1 of SEQ ID NO:90, LCDR2 of SEQ ID NO:91, and LCDR3 of SEQ ID NO:92;

m. a heavy chain variable region that contains HCDR1 of SEQ ID NO:596, HCDR2 of SEQ ID NO:597, and HCDR3 of SEQ ID NO:598; and a light chain variable region that contains LCDR1 of SEQ ID NO:612, LCDR2 of SEQ ID NO:613, and LCDR3 of SEQ ID NO:614;

n. a heavy chain variable region that contains HCDR1 of SEQ ID NO:599, HCDR2 of SEQ ID NO:600, and HCDR3 of SEQ ID NO:601; and a light chain variable region that contains LCDR1 of SEQ ID NO:615, LCDR2 of SEQ ID NO:616, and LCDR3 of SEQ ID NO:617;

o. a heavy chain variable region that contains HCDR1 of SEQ ID NO:602, HCDR2 of SEQ ID NO:603, and HCDR3 of SEQ ID NO:604; and a light chain variable region that contains LCDR1 of SEQ ID NO:618, LCDR2 of SEQ ID NO:619, and LCDR3 of SEQ ID NO:620; or

p. a heavy chain variable region that contains HCDR1 of SEQ ID NO:605, HCDR2 of SEQ ID NO:606, and HCDR3 of SEQ ID NO:607; and a light chain variable region that contains LCDR1 of SEQ ID NO:621, LCDR2 of SEQ ID NO:622, and LCDR3 of SEQ ID NO:623.

[0014] An antibody or antigen-binding fragment thereof that binds CCR7 of the present application may also contain:

a. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:13 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:29;

b. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:45 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:61;

c. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:77 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:93; or

d. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:608; and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:624.

[0015] In another embodiment, the antibody or antigen-binding fragment thereof that binds CCR7 comprises:

a. a heavy chain containing the amino acid sequence of SEQ ID NO:15 and a light chain containing the amino acid sequence of SEQ ID NO:31;

b. a heavy chain containing the amino acid sequence of SEQ ID NO:47 and a light chain containing the amino acid sequence of SEQ ID NO:63;

c. a heavy chain containing the amino acid sequence of SEQ ID NO:79 and a light chain containing the amino acid sequence of SEQ ID NO:95; or

d. a heavy chain containing the amino acid sequence of SEQ ID NO:610; and a light chain containing the amino acid sequence of SEQ ID NO:626.

[0016] An antibody or antigen-binding fragment described herein may contain one or more cysteine substitutions. In one embodiment, the antibody or antigen-binding fragment thereof contains one or more cysteine substitutions selected from S152C, S375C, or both S152C and S375C, in the heavy chain of the antibody or antigen-binding fragment, where the position numbering follows the EU system. An antibody disclosed herein may be a monoclonal antibody.

[0017] This application discloses an antibody-drug conjugate comprising the formula:

Ab-(L-(D) _m ) _n ,

or a pharmaceutically acceptable salt thereof; where

Ab is an antibody, or an antigen-binding antigen-binding fragment thereof, as disclosed herein;

L is a linker;

D is a drug moiety;

m is an integer from 1 to 8; and

n is an integer from 1 to 12.

In some embodiments, m is 1. In one embodiment, n is from about 3 to about 4. In one embodiment, the linker is selected from the group consisting of a cleavable linker, a non-cleavable linker, a hydrophilic linker, a precharged linker, and a dicarboxylic acid linker. .

[0018] In one embodiment, the linker is derived from a crosslinking reagent selected from the group consisting of N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl-4-(2-pyridyldithio)pentanoate (SPP) , N-succinimidyl-4-(2-pyridyldithio)butanoate (SPDB), N-succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB), N-succinimidyl iodoacetate (SIA), N-succinimidyl (4-iodoacetyl)aminobenzoate (SIAB), maleimide-PEG-NHS, N-succinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-sulfosuccinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (sulfo-SMCC) and 2,5- dioxopyrrolidin-1-yl-17-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5,8,11,14-tetraoxo-4,7,10,13-tetraazaheptadecan- 1-oata (CX1-1).

[0019] In other embodiments, the implementation of the linker has the following formula (IIA):

(IIA);

(IIA);

where * is connected to the thiol functional group on the antibody, and ** is connected to the thiol functional group of the fragment, which is a drug; and where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11;

X is -C(O)-NH-, -NHC(O)- or triazole; and alkylene is linear or branched.

[0020] In another embodiment, the linker has the following formula:

where y is from 1 to 11; * is linked to a thiol functional group on the antibody, and ** is linked to a thiol functional group on the drug moiety.

[0021] In one embodiment, the drug moiety is selected from the group consisting of a V-ATPase inhibitor, a proapoptotic agent, a Bcl2 inhibitor, an MCL1 inhibitor, an HSP90 inhibitor, an IAP inhibitor, an mTor inhibitor, a microtubule stabilizer, a microtubule destabilizer, auristatin , amanitin, pyrrolobenzodiazepine, RNA polymerase inhibitor, dolastatin, maytansinoid, MetAP (methionine aminopeptidase), CRM1 nuclear export inhibitor, DPPIV inhibitor, proteasome inhibitors, mitochondrial phosphoryl transfer inhibitors, protein synthesis inhibitor, kinase inhibitor, CDK2 inhibitor, CDK9 inhibitor , a kinesin inhibitor, an HDAC inhibitor, a DNA damaging agent, a DNA alkylating agent, a DNA intercalator, a DNA minor groove binding agent, and a DHFR inhibitor. In some embodiments, the cytotoxic agent is a maytansinoid, wherein the maytansinoid is N(2')-deacetyl-N(2')-(3-mercapto-l-oxopropyl)-maytansine (DM1), N(2')-deacetyl -N(2')-(4-mercapto-1-oxopentyl)-maytansine (DM3) or N(2')-deacetyl-N2-(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4) .

[0022] In one embodiment, the antibody drug conjugates disclosed herein provide the following formula (VIII):

where L ¹ represents C _1-6 alkylene, while one methylene group can be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11;

X is -C(O)-NH-, -NHC(O)- or triazole; and alkylene is linear or branched; and where n is from about 3 to about 4; or a pharmaceutically acceptable salt thereof.

[0023] In one embodiment, the antibody drug conjugates disclosed herein have the following formula:

where n is from about 3 to about 4, and Ab is an antibody containing a heavy chain containing the amino acid sequence under SEQ ID NO:47, and a light chain containing the amino acid sequence under SEQ ID NO:63; or a pharmaceutically acceptable salt thereof.

[0024] The present application also discloses a pharmaceutical composition containing the antibodies or antigen-binding fragments disclosed herein and a pharmaceutically acceptable carrier. The present application also discloses a pharmaceutical composition containing the antibody-drug conjugates disclosed herein.

[0025] The present application also discloses methods for treating or preventing cancer in a patient in need thereof, comprising administering to said patient the antibody-drug conjugates or pharmaceutical compositions disclosed herein wherein the cancer expresses CCR7.

[0026] In some embodiments of methods for treating or preventing cancer, the antibody drug conjugate or pharmaceutical composition is administered to a patient in combination with one or more additional therapeutic compounds. In one embodiment, one or more additional therapeutic compounds are selected from a standard chemotherapeutic agent, a costimulatory molecule, or a checkpoint inhibitor. In one embodiment, the costimulatory molecule is selected from OX40 agonist, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, STING or CD83 ligand. In another embodiment, the checkpoint inhibitor is selected from a PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR-beta inhibitor.

[0027] The present application also discloses antibody-drug conjugates or pharmaceutical compositions disclosed herein for use as a drug. In one embodiment, the antibody drug conjugates or pharmaceutical compositions disclosed herein are for use in the treatment or prevention of CCR7 expressing cancer in a patient in need thereof.

[0028] In one embodiment, the present application discloses the use of antibodies or antigen-binding fragments thereof, antibody-drug conjugates or pharmaceutical compositions disclosed herein for the treatment or prevention of CCR7-expressing cancer in a patient in need thereof.

[0029] In one embodiment, the present application discloses the use of antibodies or antigen-binding fragments, antibody-drug conjugates or pharmaceutical compositions disclosed herein in the manufacture of a drug.

[0030] In one embodiment, the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL), peripheral T cell lymphomas (PTCL), such as adult T cell leukemia/lymphoma (ATLL), and anaplastic large cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL) such as mantle cell lymphoma (MCL), Burkitt's lymphoma and diffuse large B cell lymphoma (DLBCL), gastric carcinomas, non-small cell lung cancer, small cell lung cancer, head and neck cancer, nasopharyngeal carcinoma (NPC) , esophageal cancer, colorectal carcinoma, pancreatic cancer, thyroid cancer, breast cancer, renal cell carcinoma, and cervical cancer. In specific embodiments, the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL), peripheral T-cell lymphomas (PTCL) such as adult T-cell leukemia/lymphoma (ATLL) and anaplastic large cell lymphoma (ALCL), non-Hodgkin's lymphoma ( NHL) such as mantle cell lymphoma (MCL), Burkitt's lymphoma and diffuse large B cell lymphoma (DLBCL), and non-small cell lung cancer.

[0031] This application also discloses nucleic acids that encode the antibodies or antigen-binding fragments disclosed herein. In one embodiment, the nucleic acid contains the nucleotide sequence under SEQ ID NO: 14, 16, 30, 32, 46, 48, 62, 64, 78, 80, 94, 96, 481, 483, 497, or 499. This application also disclosed are vectors containing nucleic acids and host cells containing vectors or nucleic acids. This application also discloses a method for obtaining the antibodies or antigen-binding fragments disclosed herein, which involves culturing the host cell and extracting the antibody from the cell culture. In one embodiment, a method for recovering an antibody from a cell culture comprises the steps of:

a) removing cells and filtering the culture;

b) carrying out purification of the culture using affinity chromatography;

c) inactivating any viruses in the culture by adjusting the pH to 3.4-3.6, then re-adjusting the pH to 5.8-6.2 and filtering the culture;

d) carrying out the purification of the culture using cation exchange chromatography and performing recovery of the culture on the column;

e) performing anion exchange chromatography on the culture;

f) removal of viruses by nanofiltration;

g) filtering the culture containing the antibody; and

h) obtaining a purified antibody.

[0032] In yet another embodiment, disclosed herein is a method for preparing an anti-CCR7 antibody drug conjugate comprising:

(a) preforming a linker-drug moiety of the following formula:

where:

the drug moiety is DM1, DM3, or DM4, and the drug moiety is attached to the linker via its thiol functionality;

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11;

X is -C(O)-NH-, -NHC(O)- or triazole; and alkylene is linear or branched;

(b) conjugating said linker-drug fragment with an antibody derived from a cell culture disclosed herein to form an antibody-drug conjugate; and

(c) performing purification of the antibody-drug conjugate.

In one embodiment, the method includes:

(a) preforming a linker-drug moiety of the following formula:

, и

, and

(b) conjugating said linker-drug fragment with an antibody derived from a cell culture disclosed herein to form an antibody-drug conjugate; and

(c) performing purification of the antibody-drug conjugate.

In another embodiment, a method for preparing an anti-CCR7 antibody drug conjugate comprises:

(a) preforming a linker-drug moiety of the following formula:

where:

the drug moiety is DM1, DM3, or DM4, and the drug moiety is attached to the linker via its thiol functionality;

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11;

X is -C(O)-NH-, -NHC(O)- or triazole; and alkylene is linear or branched;

(b) conjugating said linker drug moiety with an antibody disclosed herein to form an antibody-drug conjugate; and

(c) performing purification of the antibody-drug conjugate.

In another embodiment, a method for preparing an anti-CCR7 antibody drug conjugate comprises:

(a) preforming a linker-drug moiety of the following formula:

(b) conjugating said linker drug moiety with an antibody or antigen-binding fragment thereof disclosed herein to form an antibody-drug conjugate; and

(c) performing purification of the antibody-drug conjugate.

In another embodiment, the step of pre-forming said linker-drug moiety comprises:

a) reacting the drug moiety via its thiol functional group with:

;

;

with education:

;

;

b) the implementation of the reaction formed

с:

With:

;

;

with the formation of a fragment, which is a linker drug:

;

;

where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched; and

RG1 and RG2 are 2 reactive groups forming the X group.

In another embodiment, the step of pre-forming said linker-drug moiety comprises:

a) reacting the drug moiety via its thiol functional group with:

;

;

with education:

;

;

b) the implementation of the reaction formed

with the formation of a fragment, which is a linker drug:

In some embodiments, the antibody drug conjugate prepared according to the above methods has an average DAR, as measured by a UV spectrophotometer, of about 3 to about 4.

[0033] In another embodiment of this application, a method for preparing an anti-CCR7 antibody drug conjugate is disclosed, comprising:

(a) chemically linking the SMCC or MPET to the drug moiety, DM-1 or DM-4, to form a drug linker;

(b) conjugating said drug linker to an antibody or antigen-binding fragment thereof as disclosed herein; and

(c) performing purification of the antibody-drug conjugate.

In one embodiment, the antibody-drug conjugate prepared according to this method has an average DAR, as measured by a UV spectrophotometer, of about 3 to about 4.

[0034] This application also discloses a diagnostic reagent containing the antibody or antigen-binding fragment disclosed herein. In some embodiments, the antibody or antigen-binding fragment thereof is labeled with a radioactive label, a fluorophore, a chromophore, an imaging agent, or a metal ion.

Brief description of graphic materials

[0035] Figure 1 depicts experimental data on the in vitro ADCC activity of non-humanized and humanized anti-CCR7 antibodies in CysMab format using a surrogate ADCC reporter assay.

[0036] Figure 2 depicts experimental data on in vitro ADCC activity in non-humanized anti-CCR7 antibodies containing DAPA mutated Fc variants using a surrogate ADCC reporter assay.

[0037] Figure 3 depicts experimental data on binding to recombinant hCCR7 of anti-CCR7 antibodies in CysMab.DAPA format using an ELISA-based assay.

[0038] Figure 4A-C depicts experimental data on the functionality of the original anti-CCR7 antibodies using the β-arrestin assay in agonist mode (Figure 4A) and antagonist mode (Figure 4B, Figure 4C).

[0039] Figure 5 depicts experimental data on competition with the CCR7 ligand of anti-CCR7 antibodies in CysMab.DAPA format using FACS analysis.

[0040] Figure 6 depicts experimental epitope mapping data for parental anti-CCR7 antibodies using mutant CCR7 proteins.

[0041] Figure 7A-B depicts experimental data from piggyback ADC (pgADC) assays of parent anti-CCR7 antibodies complexed with a payload-conjugated secondary antibody fragment.

[0042] Figure 8 depicts experimental data from piggyback ADC (pgADC) cytolysis assays of the cytotoxic effects of parental 121G12 Ab complexed with a payload-conjugated secondary antibody fragment using target-negative cell lines .

[0043] Figure 9 is a graph illustrating the depletion of CD4+ and CD8a+ T cells with mouse parental CCR7 cross-reactive CysMab Ab 121G12 with wild-type Fc, either as an antibody alone or conjugated to the cytotoxin auristatin, however, these effects are eliminated by switching to the silent Fc format with DAPA mutations.

[0044] Figure 10 is a graph illustrating the dose-dependent efficacy of antibody-drug conjugates, 121G12.CysMab.DAPA.MPET.DM4 and 121G12.DAPA.sSPDB.DM4, against the KE97 xenograft model of multiple myeloma.

[0045] Figure 11 is a graph illustrating the activity of the antibody-drug conjugates, 121G12.CysMab.DAPA.MPET.DM4 and 121G12.sSPDB.DM4, in the KE97 multiple myeloma model, with dose initiation at a higher initial tumor burden than fig. ten.

[0046] Figure 12 is a graph illustrating the in vivo activity of the 121G12.CysMab.DAPA.MPET.DM4 conjugate in the HLUX1934 primary non-small cell lung tumor model.

[0047] Figure 13 is a graph illustrating the activity of the conjugated parent antibody 684E12.SMCC.DM1 in the KE97 multiple myeloma xenograft model.

[0048] Figure 14A-B depicts IHC photographs for phospho-histone H3 detection (Figure 14A) and phospho-histone H3 signal quantification (Figure 14B) for KE97 tumors 48 hours after single dose treatment with either 121G12.CysMab.DAPA. MPET.DM4 at 2, 5, or 10 mg/kg, or isotype control, IgG1.CysMab.DAPA.MPET.DM4, at 10 mg/kg, demonstrating induction of mitotic block (phospho-histone H3) after treatment with ADC to CCR7 .

[0049] Figure 15 is a graph illustrating the dose-dependent efficacy of 121G12.CysMab.DAPA.MPET.DM4 against the ABC-DLBCL OCI-LY3 xenograft model.

[0050] Figure 16 is a graph illustrating the dose-response efficacy of 121G12.CysMab.DAPA.MPET.DM4 against the GCB-DLBCL Toledo xenograft model.

[0051] Figure 17 is a graph illustrating the efficacy of 121G12.CysMab.DAPA.MPET.DM4 against the ALCL DEL xenograft model.

[0052] Figure 18 is a graph illustrating the dose-response efficacy of 121G12.CysMab.DAPA.MPET.DM4 against a patient-derived HLUX1787 xenograft NSCLC model.

Detailed description of the invention

Definitions

[0053] Unless otherwise indicated, the following terms and phrases used herein are meant to have the following meanings.

[0054] The term "alkyl" refers to a monovalent saturated hydrocarbon chain containing the specified number of carbon atoms. For example, C _1-6 alkyl refers to an alkyl group containing 1 to 6 carbon atoms. The alkyl group may be straight or branched. A typical branched alkyl group contains one, two or three branches. Examples of alkyl groups include, without limitation, methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, sec-butyl and t-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl. The term "alkylene" refers to the divalent form of "alkyl".

[0055] As used herein, the term "antibody" refers to a polypeptide from the immunoglobulin family that is capable of binding the corresponding antigen in a non-covalent, reversible, and specific manner. For example, a naturally occurring IgG antibody is a tetramer containing at least two heavy (H) chains and two light (L) chains linked together by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of a single domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) that alternate with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from amino to carboxy in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with the antigen. Antibody constant regions can mediate immunoglobulin binding to host tissues or factors, which include various cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.

[0056] The term "antibody" includes, without limitation, monoclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, and anti-idiotypic (anti-Id) antibodies (including, for example, anti-Id antibodies to antibodies of the present invention). Antibodies may be of any isotype/class (eg, IgG, IgE, IgM, IgD, IgA, and IgY) or subclass (eg, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2).

[0057] "Complementarity-determining domains" or "complementarity-determining regions" ("CDRs") refer interchangeably to VL and VH hypervariable regions. In antibody chains, the CDRs are the binding site of a target protein that confer specificity for that target protein. Each human VL or VH has three CDRs (CDR1-3, numbered sequentially from the N-terminus), representing approximately 15-20% of the variable domains. The CDRs are structurally complementary to the epitope of the target protein and are thus directly responsible for binding specificity. The remaining stretches of VL or VH, the so-called framework regions, exhibit less amino acid sequence variability (Kuby, Immunology, 4th ed., Chapter 4. W.H. Freeman & Co., New York, 2000).

[0058] The positions of CDRs and framework regions can be determined using various definitions widely known in the art, such as Kabat, Chothia, ImMunoGeneTics International Database (IMGT) (on the web at www.imgt.org/) and AbM (See, for example, Johnson et al., Nucleic Acids Res., 29:205-206 (2001); Chothia and Lesk, J. Mol. Biol., 196:901-917 (1987); Chothia et al., Nature, 342:877-883 (1989); Chothia et al., J. Mol. Biol., 227:799-817 (1992); Al-Lazikani et al ., J. Mol. Biol., 273:927- 748 (1997)). Definitions of antigen-binding active sites are also described in the following references: Ruiz et al., Nucleic Acids Res., 28:219-221 (2000); and Lefranc, MP, Nucleic Acids Res., 29:207-209 (2001); MacCallum et al., J. Mol. Biol., 262:732-745 (1996); and Martin et al., Proc. Natl. Acad. sci. USA 86:9268-9272 (1989); Martin et al., Methods Enzymol., 203:121-153 (1991); and Rees et al., in Sternberg MJE (ed.), Protein Structure Prediction, Oxford University Press, Oxford, 141-172 (1996).

[0059] Both light and heavy chains are subdivided into regions of structural and functional homology. The terms "constant" and "variable" are used in a functional sense. In this regard, it is to be understood that variable domains from both the light (VL) and heavy (VH) chain portions determine antigen recognition and specificity for it. In contrast, the light chain (CL) and heavy chain (CH1, CH2, or CH3) constant domains confer important biological properties such as secretion, placental movement, Fc receptor binding, complement binding, etc. Domain numbers are assumed to be constant region increase as they move away from the antigen-binding site or amino-terminus of the antibody. The N-terminus is the variable region and the C-terminus is the constant region; the CH3 and CL domains actually contain the heavy and light chain carboxy-terminal domains, respectively.

[0060] As used herein, the term "antigen binding fragment" refers to one or more portions of an antibody that retain the ability to specifically interact (eg, through binding, steric mismatch, stabilization/destabilization, spatial distribution) with an antigen epitope. Examples of binding fragments include, but are not limited to, single chain Fv (scFv), camelid disulfide-linked Fv (sdFv), Fab fragments, F(ab') fragments, a monovalent fragment consisting of VL, VH, CL, and CH1 domains; F(ab)2 fragment, bivalent fragment containing two Fab fragments connected by a disulfide bridge in the hinge region; Fd fragment consisting of VH and CH1 domains; Fv fragment, consisting of the VL and VH domains of one antibody arm; dAb fragment (Ward et al., Nature 341:544-546, 1989) which consists of a VH domain; as well as an isolated complementarity determining region (CDR) or other epitope-binding antibody fragments.

[0061] In addition, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be connected using recombinant methods using a synthetic linker, which provides them with a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as single-stranded Fv ("scFv"); see, e.g., Bird et al., Science 242:423-426, 1988; and Huston et al., Proc. Natl. Acad. Sci 85:5879-5883, 1988). It is envisaged that such single chain antibodies are also covered by the term "antigen binding fragment". Such antigen binding fragments are prepared using conventional techniques known to those skilled in the art and the fragments are screened for utility in the same manner as intact antibodies.

[0062] Antigen binding fragments can also be incorporated into single domain antibodies, maxitel, minitel, single domain antibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, e.g., Hollinger and Hudson, Nature Biotechnology 23: 1126-1136, 2005). Antigen binding fragments can be grafted onto polypeptide backbones such as type III fibronectin (Fn3) (see US Pat. No. 6,703,199, which describes fibronectin polypeptide monobodies).

[0063] Antigen-binding fragments can be incorporated into single-stranded molecules containing a pair of tandem Fv segments (VH-CH1-VH-CH1) which, together with complementary light chain polypeptides, form a pair of antigen-binding regions (Zapata et al., Protein Eng. 8:1057 -1062, 1995; and US Pat. No. 5,641,870).

[0064] As used herein, the term "monoclonal antibody" or "monoclonal antibody composition" refers to polypeptides comprising antibodies and antigen-binding fragments that have substantially identical amino acid sequences or are derived from the same genetic source. The term also covers preparations of antibody molecules of the same molecular composition. A monoclonal antibody composition exhibits a single binding specificity and affinity for a particular epitope.

[0065] As used herein, the term "human antibody" encompasses antibodies having variable regions in which both the framework regions and the CDRs are derived from human-derived sequences. In addition, if the antibody contains a constant region, then the constant region is also derived from such human sequences, for example, human germline sequences, or mutant variants of human germline sequences, or an antibody containing consensus framework sequences obtained by analyzing human framework sequences, for example, as described in Knappik et al., J. Mol. Biol. 296:57-86, 2000). Also contemplated are antibodies derived from human sequences where one or more CDRs have been mutated during affinity maturation or for manufacturing/payload conjugation purposes. See Kilpatrick et al., "Rapid development of affinity matured monoclonal antibodies using RIMMS", Hybridoma. 1997 Aug;16(4):381-9.

[0066] Human antibodies of the present invention may include amino acid residues that are not encoded in human sequences (e.g., mutations introduced by in vitro random or site-directed mutagenesis or by in vivo somatic mutations, or a conservative substitution that promotes stability or facilitates production).

[0067] As used herein, the term "recognize" refers to an antibody or antigen-binding fragment thereof that seeks out and interacts with (eg, binds to) its epitope, whether the epitope is linear or conformational. The term "epitope" refers to a site on an antigen to which an antibody or antigen-binding fragment of the present invention specifically binds. Epitopes can be formed by both adjacent amino acids and non-adjacent amino acids placed side by side due to the tertiary folding of the protein. Epitopes derived from contiguous amino acids are generally retained upon exposure to denaturing solvents, while epitopes derived from tertiary folding are generally lost upon exposure to denaturing solvents. An epitope typically contains at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids in a unique spatial conformation. Methods for determining the spatial conformation of epitopes include prior art techniques such as X-ray diffraction crystallography and 2D nuclear magnetic resonance (see, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, G. E. Morris, Ed. (1996)) .

[0068] As used herein, the term "affinity" refers to the strength of the interaction between an antibody and an antigen at individual antigenic sites. Within each antigenic site, the antibody "arm" variable region interacts with the antigen through weak non-covalent forces at multiple sites; moreover, the more interactions, the stronger the affinity.

[0069] The term "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigenic specificity. However, an isolated antibody that specifically binds to one antigen may be cross-reactive with other antigens. Moreover, the isolated antibody may be substantially free of other cellular material and/or chemicals.

[0070] The term "corresponding human germline sequence" refers to a nucleic acid sequence encoding a human variable region amino acid sequence or subsequence that has the highest established amino acid sequence identity with a reference amino acid sequence or variable region subsequence compared to all other all other known variable region amino acid sequences encoded in human germline immunoglobulin variable region sequences. A corresponding human germline sequence can also refer to a human variable region amino acid sequence or subsequence having the highest amino acid sequence identity with a reference variable region amino acid sequence or subsequence compared to all other variable region amino acid sequences evaluated. The corresponding human germline sequence can be framework regions only, complementarity-determining regions only, framework and complementarity-determining regions, a variable segment (as defined above), or other combinations of sequences or subsequences that constitute a variable region. Sequence identity can be determined using the methods described herein, for example, aligning two sequences using BLAST, ALIGN, or another alignment algorithm known in the art. The corresponding human germline nucleic acid or amino acid sequence may have at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity. with a reference nucleic acid sequence or variable region amino acid sequence. Corresponding human germline sequences can be determined, for example, using the international public domain ImMunoGeneTics (IMGT) database (on the web at www.imgt.org/) and V-base (on the web at vbase. mrc-cpe.cam.ac.uk).

[0071] The phrase "specifically binds" or "selectively binds", when used in the context of describing an interaction between an antigen (e.g., protein) and an antibody, antibody fragment, or binding agent derived from an antibody, refers to a binding reaction that is defining for establishing the presence of an antigen in a heterogeneous population of proteins and other biological substances, for example, in a biological sample, such as blood, serum, plasma, or a tissue sample. Thus, under certain specified immunoassay conditions, antibodies or binding agents with a particular binding specificity bind to a particular antigen at least twice as strongly as background and do not substantially bind to other antigens present in the sample. . In one embodiment, under designated immunoassay conditions, an antibody or binding agent with a particular binding specificity binds to a particular antigen at least ten (10) times stronger than background, and does not substantially bind significantly to other antigens present in the sample. Specific binding to an antibody or binding agent under such conditions may require that the antibody or agent be selected for its specificity for a particular protein. If desired or necessary, this selection can be made by discarding antibodies that cross-react with molecules from another species (eg mouse or rat) or other subtypes. Alternatively, in some embodiments, antibodies or antibody fragments are selected that cross-react with certain desired molecules.

[0072] A variety of immunoassay formats can be used to screen for antibodies that have specific immune reactivity for a particular protein. For example, ELISA immunoassays have traditionally been used to screen for antibodies having specific protein reactivity (see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for descriptions of immunoassay formats and conditions that can be used to determination of specific immune reactivity). Typically, a specific or selective binding reaction will result in a signal at least two times background, and more typically at least 10-100 times background.

[0073] The term "equilibrium dissociation constant (KD [M])" refers to the dissociation rate constant (kd [time-1]) divided by the association rate constant (ka [time-1, M-1]). Equilibrium dissociation constants can be measured using any method known in the art. Typically, antibodies of the present invention will have an equilibrium dissociation constant of less than about 10 ^-7 or 10 ^-8 M, such as less than about 10 ^-9 M or 10 ^-10 M, in some embodiments, less than about 10 ^-11 M, 10 ^{- 12} M or 10 ^-13 M.

[0074] The term "bioavailability" refers to the systemic availability (ie, blood/plasma levels) of a given amount of drug administered to a patient. Bioavailability is an absolute term referring to both the time (rate) it takes a drug to reach the general circulation from an administered dosage form and the total amount (amount) of drug in it.

[0075] As used herein, the phrase "consisting essentially of" refers to the genera or types of active pharmaceutical agents included in the method or composition, as well as any excipients that are not active in relation to the intended purpose of the methods or compositions. In some embodiments, the phrase "consisting essentially of" expressly excludes the inclusion of one or more additional active agents other than the antibody-drug conjugate of the present invention. In some embodiments, the phrase "consisting essentially of" expressly excludes the inclusion of one or more additional active agents other than the antibody-drug conjugate of the present invention and the second agent co-administered.

[0076] The term "amino acid" refers to naturally occurring, synthetic and non-natural amino acids, as well as amino acid analogs and amino acid mimetics that function similarly to naturally occurring amino acids. Naturally occurring amino acids are the amino acids encoded in the genetic code, as well as those amino acids that have been subsequently modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e. have an α-carbon that is bonded to hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide , methionine methylsulfonium. Such analogs have modified R groups (eg, norleucine) or modified peptide backbones, but retain the same basic chemical structure as the naturally occurring amino acid. Amino acid mimetics refer to chemical compounds that have a structure that differs from the general chemical structure of an amino acid, but that function similarly to a naturally occurring amino acid.

[0077] The term "conservatively modified variant" applies to both amino acid sequences and nucleic acid sequences. With respect to specific nucleic acid sequences, conservatively modified variants refer to those nucleic acids that encode identical or substantially identical amino acid sequences, or, if the nucleic acid does not encode an amino acid sequence, substantially identical sequences. Due to the degeneracy of the genetic code, any given protein is encoded by a large number of functionally identical nucleic acids. For example, all of the codons GCA, GCC, GCG, and GCU code for the amino acid alanine. Thus, at each position at which the codon is given by alanine, the codon can be changed to any of the corresponding codons described without changing the encoded polypeptide. Such nucleic acid variations are "silent variations", which are one kind of variation with conservative modifications. In this document, each nucleic acid sequence that encodes a polypeptide also describes each possible silent variation of the nucleic acid. One skilled in the art will appreciate that every codon in a nucleic acid (with the exception of AUG, which is usually the only codon for methionine, and TGG, which is usually the only codon for tryptophan) can be modified to produce a functionally identical molecule. Accordingly, each silent variation of a nucleic acid that encodes a polypeptide is implicitly defined in each described sequence.

[0078] In the case of polypeptide sequences, "conservatively modified variants" encompass individual substitutions, deletions, or additions in a polypeptide sequence that result in the substitution of an amino acid for a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to, and not exclusive to, the polymorphic variants, cross-species homologues and alleles of the present invention. The following eight groups contain amino acids that are conservative substitutions for each other: 1) alanine (A), glycine (G); 2) aspartic acid (D), glutamic acid (E); 3) asparagine (N), glutamine (Q); 4) arginine (R), lysine (K); 5) isoleucine (I), leucine (L), methionine (M), valine (V); 6) phenylalanine (F), tyrosine (Y), tryptophan (W); 7) serine (S), threonine (T); and 8) cysteine (C), methionine (M) (see, for example, Creighton, Proteins (1984)). In some embodiments, the term "conservative sequence modifications" is used to refer to amino acid modifications that do not significantly affect or alter the binding characteristics of an antibody comprising an amino acid sequence.

[0079] As used herein, the term "optimized" refers to a nucleotide sequence encoding an amino acid sequence that has been altered using codons preferred in a producing cell or organism, typically a eukaryotic cell, e.g., a yeast cell, a Pichia cell, a fungus cell, Trichoderma cell, Chinese hamster ovary (CHO) cell, or human cell. The optimized nucleotide sequence is designed in such a way as to fully or as far as possible retain the amino acid sequence originally encoded in the original nucleotide sequence, which is also known as the "original" sequence.

[0080] The terms "percent identity" or "percent identity", in the context of two or more nucleic acids or polypeptide sequences, refers to the degree to which two or more sequences or subsequences are the same. Two sequences are "identical" if they have the same amino acid or nucleotide sequence over the region to be compared. Two sequences are "substantially identical" if two sequences have a specified percentage of amino acid residues or nucleotides that are the same (i.e. 60% identity, optionally 65%, 70%, 75%, 80%, 85%, 90% , 95% or 99% identity over the specified region or, if not specified, throughout the entire sequence), when compared and aligned to ensure maximum agreement throughout the comparison window or specified region, as measured using one of the following sequence comparison algorithms, or through manual alignment and visual inspection. Optionally, identity exists over a region that is at least about 30 nucleotides (or 10 amino acids) long, or more preferably over a region that is 100-500 or 1000 or more nucleotides (or 20, 50, 200 or more amino acids) long. ).

[0081] When comparing sequences, typically one sequence acts as a reference sequence against which the test sequences are compared. When using the sequence comparison algorithm, the test and reference sequences are entered into the computer, if necessary, the coordinates of the subsequences are set, and the software parameters of the algorithm for sequence analysis are set. Default program settings may be used, or alternative settings may be set. Based on the program parameters, the sequence comparison algorithm then calculates percent sequence identity values for the test sequences relative to the reference sequence.

[0082] As used herein, a "comparison window" refers to a segment of any number of contiguous positions selected from the group consisting of 20 to 600, typically about 50 to about 200, more typically about 100 to about 150, in which the sequence can be compared to a reference sequence with the same number of contiguous positions after the two sequences have been optimally aligned. Methods for aligning sequences for comparison are well known in the art. Optimal alignment of sequences for comparison can be performed, for example, using the Smith-Waterman Local Homology Search Algorithm, Adv. Appl. Math. 2:482c (1970), using the Needleman-Wunsch homology region alignment algorithm, J. Mol. Biol. 48:443 (1970), using the Pearson-Lipman similarity search method, Proc. Natl. Acad. sci. USA 85:2444 (1988), using computer implementations of such algorithms (GAP, BESTFIT, FASTA, and TFASTA as part of the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Dr., Madison, WI) or using manual alignment and visual browsing (see, for example, Brent et al., Current Protocols in Molecular Biology, 2003).

[0083] Two examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402, 1977; and Altschul et al., J. Mol. biol. 215:403-410, 1990, respectively. BLAST analysis software is available from the National Center for Biotechnology Information. At the first stage, this algorithm involves the identification of pairs of sequences with a high score of similarity (HSP) by identifying short words of length W in the requested sequence that either match or satisfy some threshold score T with a positive value when aligned with a word of the same length in the sequence of Database. T is known as the neighborhood word threshold score (Altschul et al., supra). These initial neighbor word matches act as "seeds" to start searches designed to find longer HSPs containing them. Word matches are extended in both directions along each sequence for as long as the total alignment score can increase. For nucleotide sequences, total scores are calculated using the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatched residues; always <0). In the case of amino acid sequences, a substitution matrix is used to calculate the total score. The extension of word matches in each direction stops when the total alignment score decreases by X from its maximum achieved value; the total score tends to zero or lower due to the accumulation of one or more negative-scoring residual alignments; or the end of either sequence is reached. The W, T and X parameters of the BLAST algorithm determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) defaults to a word length (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program defaults to a word length of 3 and an expectation (E) of 10 and a BLOSUM62 substitution matrix (see Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915), alignments (B) of 50, wait (E) of 10, M=5, N=-4, and comparison of both strands.

[0084] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873-5787, 1993). One measure of similarity provided in the BLAST algorithm is the smallest sum probability (P(N)), which indicates the probability that a match between two nucleotide or amino acid sequences occurred by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability when comparing the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.

[0085] Percent identity between two amino acid sequences can also be determined using the algorithm from E. Meyers and W. Miller (Comput. Appl. Biosci. 4:11-17, (1988) which was included in the ALIGN program (version 2.0), using a PAM120 residue substitution weight table, a gap extension penalty of 12, and a gap opening penalty of 4. In addition, percent identity between two amino acid sequences can be determined using the Needleman-Wunsch algorithm, J. Mol. Biol. 48:444-453, (1970), which was included in the GAP program as part of the GCG software package (available at www.gcg.com), using either the BLOSUM62 matrix or the PAM250 matrix, and also a gap opening penalty of 16, 14, 12, 10, 8, 6, or 4, and a gap extension penalty of 1, 2, 3, 4, 5, or 6.

[0086] In addition to the percent sequence identity mentioned above, another indication that two nucleic acid or polypeptide sequences are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with antibodies induced by the polypeptide, encoded by the second nucleic acid, as described below. Thus, the polypeptide is typically substantially identical to the second polypeptide, for example, if the two peptides differ only in conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules, or their complementary strands, hybridize to each other under stringent conditions, as described below. Another indication that two nucleic acid sequences are substantially identical is that the same primers can be used to amplify the sequence.

[0087] The term "nucleic acid" is used herein interchangeably with the term "polynucleotide" and refers to deoxyribonucleotides or ribonucleotides and polymers based on them in single or double stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages that are synthetic, naturally occurring or non-naturally occurring, that have binding properties similar to a reference nucleic acid, and that are metabolized like reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methylphosphonates, chiral methylphosphonates, 2-O-methylribonucleotides, peptidonucleic acids (PNAs).

[0088] Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses its conservatively modified variants (eg, substitutions based on degenerate codons) and complementary sequences, as well as the sequence specified explicitly. Namely, as detailed below, substitutions based on degenerate codons can be performed by creating sequences in which the third position in one or more selected (or all) codons is replaced by any of the canonical nucleosides and/or a deoxyinosine residue (Batzer et al. , Nucleic Acid Res. 19:5081, 1991; Ohtsuka et al., J. Biol. Chem. 260:2605-2608, 1985; and Rossolini et al., Mol. Cell. Probes 8:91-98, 1994).

[0089] In the context of nucleic acids, the term "operably linked" refers to the functional relationship of two or more segments of a polynucleotide (eg, DNA). Generally, it refers to the functional relationship of a transcriptional regulatory sequence to a transcribed sequence. For example, a promoter or enhancer sequence is operably linked to a coding sequence if it stimulates or modulates the transcription of the coding sequence in an appropriate host cell or other expression system. Typically, transcription-regulating promoter sequences that are operably linked to the transcribed sequence are physically contiguous to the transcribed sequence, i.e. they function in the cis position. However, some transcriptional control sequences, such as enhancers, need not be physically contiguous or in close proximity to the coding sequences whose transcription they enhance.

[0090] The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to polymers of amino acids in which one or more amino acid residues is an artificial chemical mimetic of the corresponding naturally occurring amino acid, as well as to polymers of naturally occurring amino acids and a polymer of non-naturally occurring amino acids. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

[0091] As used herein, the term "antibody drug conjugate" or "immunoconjugate" refers to the association of an antibody or antigen-binding fragment thereof with another agent, such as a chemotherapeutic agent, toxin, immunotherapeutic agent, imaging probe, and the like. The bond may be covalent bonds or non-covalent interactions such as through electrostatic forces. Various linkers known in the art can be used to form an antibody-drug conjugate. In addition, the antibody-drug conjugate may be provided in the form of a fusion protein that can be expressed from a polynucleotide encoding an immunoconjugate. As used herein, a "fusion protein" refers to proteins created by combining two or more genes or gene fragments that originally encoded separate proteins (including peptides and polypeptides). Translation of the fused gene results in a single protein with functional properties derived from each of the original proteins.

[0092] The term "subject" encompasses humans and non-human animals. Non-human animals include all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians and reptiles. Except where noted, the terms "patient" or "subject" are used interchangeably herein.

[0093] As used herein, the term "cytotoxin" or "cytotoxic agent" refers to any agent that is detrimental to cell growth and proliferation and that can function to reduce, inhibit, or destroy a cell or cancer.

[0094] As used herein, the term "anti-cancer agent" refers to any agent that can be used to treat or prevent a disorder of cell proliferation such as cancer, including, without limitation, cytotoxic agents, chemotherapeutic agents, radiotherapy, and radiotherapy agents. therapy, targeting anticancer agents and immunotherapeutic agents.

[0095] As used herein, the term "drug moiety" or "payload" refers to a chemical moiety that is conjugated to an antibody or antigen-binding moiety of the present invention, and may encompass any therapeutic or diagnostic agent, such as an anticancer, anti-inflammatory, anti-infective (eg, antifungal, antibacterial, antiparasitic, antiviral), or anesthetic. For example, a drug moiety may be an anticancer agent such as a cytotoxin. In some embodiments, the drug moiety is selected from V-ATPase inhibitor, HSP90 inhibitor, IAP inhibitor, mTor inhibitor, microtubule stabilizer, microtubule destabilizer, auristatin, dolastatin, maytansinoid, MetAP (methionine aminopeptidase), RNA polymerase inhibitor, pyrrolobenzodiazepine (PBD), amanitin, CRM1 nuclear export inhibitor, DPPIV inhibitor, mitochondrial phosphoryl transfer inhibitor, protein synthesis inhibitor, kinase inhibitor, CDK2 inhibitor, CDK9 inhibitor, proteasome inhibitor, kinesin inhibitor, HDAC inhibitor, DNA damaging agent, DNA an alkylating agent, a DNA intercalator, a DNA minor groove binding agent, and a DHFR inhibitor. Methods for attaching each of these to an antibody-compatible linker and method of the present invention are known in the art. See, for example, Singh et al., (2009) Therapeutic Antibodies: Methods and Protocols, vol. 525, 445-457. In addition, the payload may be a biophysical detection probe, a fluorophore, a spin label, an infrared detection probe, an affinity detection probe, a chelator, a spectroscopic detection probe, a radioactive probe, a lipid molecule, polyethylene glycol, a polymer, a spin label, DNA, RNA, protein, peptide, surface, antibody, antibody fragment, nanoparticle, quantum dot, liposome, PLGA particle, saccharide or polysaccharide.

[0096] The term "maytansinoid drug moiety" refers to the portion of an antibody-drug conjugate that has the structure of a maytansinoid compound. First, maytansine was isolated from the bush Maytenus serrata, growing in East Africa (US patent No. 3896111). Subsequently, it was found that some microorganisms also produce maytansinoids, such as maytansinol and esters of C-3-maytansinol (US patent No. 4151042). Synthetic maytansinol and maytansinol analogues have been reported. See U.S. Patent Nos. 4,137,230; 4248870; 4256746; 4260608; 4265814; 4294757; 4307016; 4308268; 4308269; 4309428; 4313946; 4315929; 4317821; 4322348; 4331598; 4361650; 4364866; 4424219; 4450254; 4362663 and 4371533, and Kawai et al., (1984) Chem. Pharm. Bull. 3441-3451), each of which is expressly incorporated by reference. Examples of specific maytansinoids useful for conjugation include DM1, DM3 and DM4.

[0097] "Tumor" refers to the growth and proliferation of neoplastic cells, whether malignant or benign, and all precancerous and cancerous cells and tissues.

[0098] The term "antitumor activity" means a reduction in the rate of proliferation, viability or metastatic activity of tumor cells. For example, antitumor activity can be shown by a drop in the rate of growth of abnormal cells that occurs during therapy, or stabilization or reduction in tumor size, or longer survival due to therapy compared to a control without therapy. Such activity can be assessed using accepted in vitro or in vivo tumor models, including, but not limited to, xenograft models, allogeneic graft models, MMTV models, and other known models known in the art for testing antitumor activity.

[0099] The term "malignant tumor" refers to a benign tumor or cancer. As used herein, the term "cancer" encompasses a malignant tumor that is characterized by deregulated or uncontrolled cell growth. Exemplary types of cancer include: carcinomas, sarcomas, leukemias, and lymphomas.

[00100] The term "cancer" encompasses primary malignant tumors (e.g., tumors whose cells have not migrated to locations in the subject's body other than the site of the original tumor) and secondary malignant tumors (e.g., tumors that have developed by metastasis, migration of tumor cells during secondary sites that are different from the site of the original tumor).

[00101] The term "CCR7" (also known as BLR2, CC-CKR-7, CCR-7, CD197, CDw197, CMKBR7, EBI1, or C-C motif chemokine receptor 7) refers to a member of the G protein-coupled receptor family. Последовательность нуклеиновой кислоты и аминокислотная последовательность CCR7 человек была опубликована в GenBank под следующими №№ доступа: NP_001829, NP_001288643, NP_001288645, NP_001288646, NP_001288647 (аминокислотные последовательности) и NM_001838, NM_001301714, NM_001301716, NM_001301717, NM_001301718 (нуклеотидные последовательности). As used herein, the term "CCR7" is used collectively to refer to all naturally occurring isoforms of the CCR7 protein or a variant thereof.

[00102] The term "variant" refers to a polypeptide that has a substantially identical amino acid sequence to a reference polypeptide, or is encoded by a substantially identical nucleotide sequence, and may have one or more of the activities of the reference polypeptide. For example, a variant may have approximately 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity with a reference polypeptide, while retains one or more activities of the reference polypeptide.

[00103] As used herein, the terms "treat," "treat," or "treat" any disease or disorder, in one embodiment, refer to lessening the severity of the disease or disorder (i.e., slowing, or stopping, or reducing the progression of the disease, or at least one of its clinical symptoms). In another embodiment, "treat", "treating", or "treatment" refers to alleviating or reducing the severity of at least one physical parameter, including those that may not be apparent to the patient. In yet another embodiment, "treating", "administering treatment", or "treatment" means modulating a disease or disorder, either physically (eg, stabilizing an overt symptom), physiologically (eg, stabilizing a physical parameter), or both.

[00104] As used herein, the term "prevent", "prevent" or "prevention" of any disease or disorder refers to the prophylactic treatment of a disease or disorder or to slow the onset or progression of a disease or disorder.

[00105] The term "therapeutically acceptable amount" or "therapeutically effective dose" interchangeably refers to an amount sufficient to produce the desired result (i.e., reduction in tumor size, inhibition of tumor growth, prevention of metastasis, inhibition or prevention of viral, bacterial, fungal or parasitic infection). In some embodiments, the therapeutically acceptable amount does not induce or cause unwanted side effects. In some embodiments, the therapeutically acceptable amount induces or causes side effects, but only those that are acceptable from the point of view of medical professionals, given the condition of the patient. A therapeutically acceptable amount can be determined by first administering a low dose and then gradually increasing this dose until the desired effect is achieved. A "prophylactically effective dose" and a "therapeutically effective dose" of the molecules of the present invention may prevent the onset of or, respectively, result in a reduction in the severity of symptoms of a disease, including symptoms associated with cancer.

[00106] The term "co-administration" refers to the presence of two active agents in the blood of an individual. Active agents that are co-administered may be delivered simultaneously or sequentially.

[00107] The present invention provides antibodies, antibody fragments (eg, antigen-binding fragments), and drug conjugates thereof, i. antibody drug conjugates or ADCs that bind to CCR7. In particular, the present invention provides antibodies and antibody fragments (eg, antigen-binding fragments) that bind to CCR7 and are internalized after such binding. The antibodies and antibody fragments (eg, antigen-binding fragments) of the present invention can be used to prepare antibody-drug conjugates. In addition, the present invention provides antibody-drug conjugates that have the desired pharmacokinetic characteristics and other desired properties, and thus can be used to treat or prevent cancer expressing CCR7. The present invention further provides pharmaceutical compositions containing the antibody-drug conjugates of the present invention; as well as methods of making and using such pharmaceutical compositions for the treatment or prevention of cancer.

Antibody drug conjugates

[00108] The present invention provides antibody-drug conjugates, also referred to as immunoconjugates, wherein an antibody, antigen-binding fragment, or functional equivalent thereof that specifically binds to CCR7 is linked to a drug moiety. In one aspect, the antibodies, antigen-binding fragments, or functional equivalents thereof of the present invention are linked via a covalent bond through a linker to a drug moiety that is an anticancer agent. The antibody-drug conjugates of the present invention can deliver an effective dose of an anticancer agent (eg, a cytotoxic agent) to tumor tissue expressing CCR7, whereby greater selectivity (and lower effective dose) can be achieved.

[00109] In one aspect of the present invention, an immunoconjugate of formula (I) is provided:

Ab-(L-(D) _m ) _n ,

where Ab is a CCR7-binding antibody described in this document;

L is a linker;

D is a drug moiety;

m is an integer from 1 to 8; and

n is an integer from 1 to 20. In one embodiment, n is an integer from 1 to 10, 2 to 8, or 2 to 5. In a specific embodiment, n is 2, 3, or 4. In some embodiments, m is one; in other embodiments, m is 2, 3, or 4.

[00110] Although the ratio of drug to antibody has an exact value in a particular conjugate molecule (e.g., n times m in formula (I)), it is known that the value will often be an average when used to describe a sample containing many molecules due to some degree of heterogeneity usually associated with the conjugation step. The average loading for an immunoconjugate sample is referred to herein as the drug to antibody ratio or "DAR". In some embodiments, when the drug is a maytansinoid, it is referred to as "MAR." In some embodiments, the DAR is from about 2 to about 6, and is typically about 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7.0, 7.5, 8 ,0. In some embodiments, at least 50% of the sample, by weight, is a compound having an average DAR of plus or minus 2, and preferably at least 50% of the sample is a conjugate that has an average DAR of plus or minus 1. Embodiments encompass immunoconjugates in which DAR is approximately 3.5, 3.6, 3.7, 3.8 or 3.9. In some embodiments, a DAR of "about n" means that the measured DAR value is within 20% of n.

[00111] The present invention is also directed to immunoconjugates containing antibodies, antibody fragments (eg, antigen-binding fragments) and their functional equivalents disclosed herein, connected or conjugated to a drug moiety. In one embodiment, drug moiety D is a maytansinoid drug moiety, including those having the structure:

where the wavy line indicates the covalent bond of the sulfur atom of the maytansinoid to the linker of the antibody-drug conjugate. R is at each occurrence independently H or C ₁ -C ₆ alkyl. The alkylene chain attaching the amide group to the sulfur atom may be methanyl, ethanyl or propyl, i.e. r is 1, 2 or 3. (U.S. Patent No. 633410, U.S. Patent No. 5208020, Chari et al. (1992) Cancer Res. 52;127-131, Lui et al. (1996) Proc. Natl. Acad. Sci. 93:8618-8623).

[00112] All stereoisomers of the maytansinoid drug moiety are contemplated for use in the immunoconjugates of the present invention, i.e., any combination of the R and S configurations at the chiral carbons of the maytansinoid. In one embodiment, the maytansinoid drug moiety has the following stereochemistry.

[00113] In one embodiment, the maytansinoid drug moiety is N 2'-deacetyl-N ² '-( ³ -mercapto-1-oxopropyl)-maytansine (also known as DM1). DM1 is represented by the following structural formula.

DM1

[00114] In another embodiment, the maytansinoid drug moiety is N 2'-deacetyl- N ² ' ^- (4-mercapto-1-oxopentyl)-maytansine (also known as DM3). DM3 is represented by the following structural formula.

DM3

[00115] In another embodiment, the maytansinoid drug moiety is N 2'-deacetyl-N ² ' ^- (4-methyl-4-mercapto-1-oxopentyl)-maytansine (also known as DM4). DM4 is represented by the following structural formula.

DM4

[00116] A drug moiety D can be linked to an antibody Ab using a linker L. L is any chemical moiety capable of linking a drug moiety to an antibody through covalent bonds. A crosslinking reagent is a bifunctional or multifunctional reagent that can be used to join a drug moiety and an antibody to form antibody-drug conjugates. Antibody-drug conjugates can be prepared using a cross-linking reagent having reactive functional groups capable of binding to both the drug moiety and the antibody. For example, a cysteine, thiol, or amine, for example, at the N-terminus or side chain of an amino acid such as lysine, in an antibody can form a bond with a functional group of a crosslinker. Alternatively, antibody-drug conjugates can be prepared by first forming a linker-drug moiety (or a linker-drug moiety, both terms are used interchangeably) and reacting the linker-drug moiety agent with an antibody. In some cases, the linker moiety is gradually assembled on the drug by several connecting moieties until the desired linker-drug moiety is obtained.

[00117] In one embodiment, L is a cleavable linker. In another embodiment, L is a non-cleavable linker. In some embodiments, L is an acid-labile linker, a photo-labile linker, a peptidase-cleavable linker, an esterase-cleavable linker, a disulfide-cleavable linker, a hydrophilic linker, a pre-charged linker, a glycosidase-cleavable linker, a phosphodiesterase-cleavable linker, a phosphatase-cleavable linker, or a dicarboxylic linker. acids.

[00118] Suitable cross-linking reagents that form a non-cleavable linker between a drug moiety, such as a maytansinoid, and an antibody are well known in the art and can form non-cleavable linkers that contain a sulfur atom (such as SMCC) or linkers that do not contain a sulfur atom. Preferred crosslinking reagents that form non-cleavable linkers between a drug moiety, such as a maytansinoid, and an antibody, contain a maleimide or haloacetyl moiety. In accordance with the present invention, such non-cleavable linkers are said to be derived from maleimide or haloacetyl moieties.

[00119] Cross-linking reagents containing a maleimide moiety include, but are not limited to, N-succinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC), N- succinimidyl-4-(maleimidomethyl)cyclohexane-1-carboxy-(6-amidocaproate), which is the "long chain" analogue of SMCC (LC-SMCC), κ-maleimidoundecanoic acid N-succinimidyl ester (KMUA), N-succinimidyl complex γ-maleimidobutyric acid ester (GMBS), ε-maleimidocaproic acid N-succinimidyl ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(α-maleimidoacetoxy)-succinimide ester (AMSA), succinimidyl-6-(β-maleimidopropionamido)hexanoate (SMPH), N-succinimidyl-4-(p-maleimidophenyl)-butyrate (SMPB), N-(-p-maleimidophenyl)isocyanate (PMIP) and maleimide-based crosslinkers, containing a polyethylene glycol spacer such as MAL-PEG-NHS. These crosslinkers form non-cleavable linkers derived from maleimide-based moieties. Representative structures of maleimide-based crosslinkers are shown below.

[00120] In another embodiment, linker L is derived from N -succinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC), or MAL-PEG-NHS.

[00121] Cross-linking reagents containing a haloacetyl-based moiety include N-succinimidyl iodoacetate (SIA), N-succinimidyl (4-iodoacetyl) aminobenzoate (SIAB), N-succinimidyl bromoacetate (SBA), and N-succinimidyl-3-(bromoacetamido) propionate (SBAP). These crosslinkers form a non-cleavable linker derived from haloacetyl-based moieties. Representative structures of haloacetyl crosslinkers are shown below.

или

or

[00122] In one embodiment, linker L is from N-succinimidyliodoacetate (SIA) or N-succinimidyl(4-iodoacetyl)aminobenzoate (SIAB).

[00123] Suitable crosslinking reagents that form a cleavable linker between a drug moiety, such as a maytansinoid, and an antibody, are well known in the art. Disulfide-containing linkers are linkers cleavable by disulfide exchange, which can occur under physiological conditions. In accordance with the present invention, such cleavable linkers are said to be derived from disulfide-based moieties. Suitable disulfide crosslinkers include N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl-4-(2-pyridyldithio)pentanoate (SPP), N-succinimidyl-4-(2-pyridyldithio)butanoate ( SPDB) and N -succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB), the structures of which are shown below. These disulfide crosslinkers form a cleavable linker derived from disulfide-based moieties.

N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP),

N-succinimidyl-4-(2-pyridyldithio)pentanoate (SPP),

N-succinimidyl-4-(2-pyridyldithio)butanoate (SPDB) and

N -succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB).

[00124] In one embodiment, linker L is derived from N-succinimidyl-4-(2-pyridyldithio)butanoate (SPDB).

[00125] Suitable crosslinkers that form a charged linker between a drug moiety, such as a maytansinoid, and an antibody are known as precharged crosslinkers. In one embodiment, the linker L is from a pre-charged CX1-1 crosslinker. The structure of CX1-1 is shown below.

2,5-dioxopyrrolidin-1-yl-17-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5,8,11,14-tetraoxo-4,7,10, 13-tetraazaheptadecan-1-oate (CX1-1)

[00126] Each of the crosslinkers depicted above has an NHS ester at one end that reacts with the primary amine of the antibody to form an amide bond, and at the other end a maleimide group or a pyridinyl disulfide group that reacts with the sulfhydryl of the moiety representing is a maytansinoid drug, with the formation of a thioether or disulfide bond.

[00127] In another embodiment, suitable crosslinking moieties that form a cleavable linker between the drug (e.g., maytansinoid) moiety and the antibody are represented by the following formula (II):

(II);

(II);

where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched.

In one aspect of this embodiment, y is 5, 7, 9, or 11. In another aspect of this embodiment, y is less than 5.

In yet another embodiment, suitable crosslinking moieties according to Formula I are selected from the group consisting of:

;

;

where y is between 1 and 11.

[00128] In the case of the crosslinking moieties depicted above (i.e., MBT, MPET, MEPET; MMTBT, MPPT, MPBT), the maleimide group allows reaction with the cysteine sulfhydryl (or thiol) in the antibody, thereby forming a thioether communications; and the thiol functional group of the crosslinking moiety combines with the thiol of the maytansinoid drug moiety to form a cleavable disulfide bond. Due to the cross-reactive nature of the linking moiety of formula (I) (thiol and maleimide can cross-react), one skilled in the art will recognize that the linking moiety must assemble progressively onto the drug moiety as depicted in Scheme 1 .

[00129] In accordance with the above embodiment, linkers derived from crosslinking fragments (i.e. MBT, MPET, MEPET) can be depicted as follows:

(IIA);

(IIA);

where * is connected to a thiol functional group on the antibody, and ** is connected to the thiol functional group of a drug moiety (eg, a maytansinoid drug moiety, DM1, DM3, or DM4).

[00130] In accordance with the above embodiment, linkers derived from crosslinking fragments (i.e. MBT, MPET, MEPET) can be depicted as follows:

where y is from 1 to 11; * linked to a thiol functional group on an antibody, and ** linked to a thiol functional group on a drug moiety (eg, maytansinoid drug, DM1, DM3, or DM4).

[00131] In a preferred embodiment, the linker has the following formula:

;

;

where * is connected to the thiol functional group on the antibody, and ** is connected to the thiol functional group of the maytansinoid drug (DM1, DM3 or DM4).

[00132] In one embodiment, the present invention provides a linker drug moiety with the formula:

; где

; where

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched.

[00133] In another embodiment, the present invention relates to the gradual formation of the above linker-drug conjugate disclosed in Scheme 1 herein.

[00134] In one embodiment, the present invention provides compounds of the linker drug moiety having one of the following formulas (III), (IV) and (V):

where L ¹ represents C _1-6 alkylene, while one methylene group can be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched.

[00135] In one embodiment, the present invention provides compounds of the linker drug moiety selected from the following formulae:

;

;

;

;

и

and

.

.

[00136] In another embodiment, the present invention provides compounds of the linker drug moiety selected from the following formulae:

[00137] In another embodiment, the linker drug of the present invention is any of the following formulae:

где y составляет от 1 до 11, предпочтительно от 1 до 5.

where y is from 1 to 11, preferably from 1 to 5.

[00138] In one embodiment, the linker drug of the present invention is represented by any of the following structural formulas:

[00139] In one embodiment, the conjugate of the present invention is represented by any of the following structural formulas:

,

,

where:

Ab is an antibody or antigen-binding fragment thereof that specifically binds CCR7;

n, which indicates the number of linker-drug groups (L-D-) linked to the Ab by forming an amide bond with a primary amine on the Ab, is an integer from 1 to 20. In one embodiment, n is an integer from 1 to 10 , 2 to 8, or 2 to 5. In a particular embodiment, n is 3 or 4.

[00140] In another embodiment, the conjugate of the present invention is any of the following formulas (VI), (VII) and (VIII):

;

;

where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched; and

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of linker-drug groups (L-D-) linked to the Ab by forming an amide bond with a primary amine on the Ab, is an integer from 1 to 20. In one embodiment, n is an integer from 1 to 10 , 2 to 8, or 2 to 5. In a particular embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00141] In another embodiment, the conjugate of the present invention has formula (VIA) or (VIB) corresponding to open forms of the succinimide conjugate of formula (VI):

where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched; and

Ab is an antibody or antigen binding fragment;

n, which indicates the number of linker-drug groups (L-D-) linked to the Ab by forming an amide bond with a primary amine on the Ab, is an integer from 1 to 20. In one embodiment, n is an integer from 1 to 10 , 2 to 8, or 2 to 5. In a particular embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00142] In another embodiment, the conjugate of the present invention has formula (VIIA) or (VIIB) corresponding to open forms of the succinimide conjugate of formula (VII):

where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched; and

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of linker-drug groups (L-D) linked to the Ab by forming an amide bond with a primary amine on the Ab, is an integer from 1 to 20. In one embodiment, n is an integer from 1 to 10, 2 to 8 or 2 to 5. In a particular embodiment, n is 3 or 4. In some embodiments, the antibody or antigen-binding fragment thereof binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00143] In another embodiment, the conjugate of the present invention has formula (VIIIA), (VIIIB) corresponding to open forms of the succinimide conjugate of formula (VIII):

where:

L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched; and

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of linker-drug groups (L-D-) linked to the Ab by forming an amide bond with a primary amine on the Ab, is an integer from 1 to 20. In one embodiment, n is an integer from 1 to 10 , 2 to 8, or 2 to 5. In a particular embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00144] In one embodiment, each antibody drug conjugate disclosed herein, wherein the drug linker moiety is attached to the antibody via succinimide, can also exist as open forms of succinimide, as generally depicted in formulas (VIA), (VIB), (VIIA), (VIIB), (VIIIA) and (VIIIB).

[00145] In another embodiment, the conjugate of the present invention is represented by any of the following structural formulas:

where:

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of D-L groups linked to the Ab by forming a thioether bond with the sulfhydryl of Ab, is an integer from 1 to 12, or from 1 to 8, or preferably from 1 to 4. In a specific embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00146] In another embodiment, the conjugate of the present invention is represented by any of the following structural formulas:

, и

, and

;

;

as well as the corresponding open forms of succinimide;

where:

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of D-L groups linked to the Ab by forming a thioether bond with the sulfhydryl of Ab, is an integer from 1 to 12, or from 1 to 8, or preferably from 1 to 4. In a specific embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00147] In another embodiment, the conjugate of the present invention is represented by any of the following structural formulas:

;

;

as well as the corresponding open forms of succinimide;

where:

y is from 1 to 11, preferably from 1 to 5;

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of D-L groups linked to the Ab by forming a thioether bond with the sulfhydryl of Ab, is an integer from 1 to 12, or from 1 to 8, or preferably from 1 to 4. In a specific embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00148] In another embodiment, the conjugate of the present invention is represented by the following formulas:

as well as the corresponding open forms of succinimide;

where:

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of D-L groups linked to the Ab by forming a thioether bond with the sulfhydryl of Ab, is an integer from 1 to 12, or from 1 to 8, or preferably from 1 to 4. In a specific embodiment, n is 3 or 4. In some embodiments, the antibody, or antigen-binding fragment thereof, binds to an antigen that is expressed on tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00149] In a preferred embodiment, the conjugate of the present invention is represented by the following formula:

where:

Ab is an antibody or antigen-binding fragment thereof;

n, which indicates the number of D-L groups bonded to the Ab by forming a thioether bond with the sulfhydryl Ab, is an integer from 1 to 20. In some embodiments, n is an integer from 1 to 12. In some embodiments, n is an integer from 1 to 8. In some embodiments, n is an integer from 1 to 4. In a specific embodiment, n is 3 or 4. In another embodiment, the average value of n is from about 3 to about 4. In some embodiments, the antibody or its the antigen-binding moiety binds to an antigen that is expressed on the tumor cells. In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds CCR7. In other embodiments, the antibody, or antigen-binding fragment thereof, specifically binds P-cadherin, cadherin 6, FGFR2, or FGFR4.

[00150] In one embodiment, the average molar ratio of the drug (e.g., DM1, DM3 or DM4) to the antibody in the conjugate (i.e. the mean n, also known as the maytansinoid to antibody ratio (MAR), is about 1 to about 10, about 2 to about 8 (e.g., 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3, 1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5, 6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, or 8 ,1), about 2.5 to about 7, about 3 to about 5, about 2.5 to about 4.5 (e.g., about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5) , from about 3.0 to about 4.0, from about 3.2 to about 4.2, or from about 4.5 to 5.5 (for example, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4 or approximately 5.5).

[00151] In one aspect of the present invention, the conjugate of the present invention is substantially highly pure and has one or more of the following: (a) greater than about 90% (e.g., greater than or equal to about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%), preferably greater than about 95% of the conjugate species are monomeric, (b) the level of unconjugated linker in the conjugate preparation is less than about 10% (for example, less than or equal to about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0%) (relative to total linker), (c) less than 10% of the conjugate species are stitched (eg. less than or equal to about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0%), (d) free drug level (e.g., DM1, DM3, or DM4) in a conjugate preparation is less than about 2% (e.g., less than or equal to approximately 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0 .6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, or 0%) (mol/mol relative to total cytotoxic agent); and/or (e) there is no significant increase in free drug levels (e.g., DM1, DM3 or DM4) after storage (for example, after about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 1 year, about 2 years, about 3 years, about 4 years or approximately 5 years). A "significant increase" in the level of free drug (e.g., DM1, DM3 or DM4) means that after some storage time (for example, approximately 1 week approximately 2 weeks approximately 3 weeks approximately 1 month approximately 2 months approximately 3 months approximately 4 months approximately 5 months approximately 6 months approximately 1 year approximately 2 years approximately 3 years approximately 4 years or approximately 5 years) an increase in the level of free drug (for example, DM1, DM3 or DM4) is less than about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1 .6%, approximately 1.7%, approximately 1.8%, approximately 1.9%, approximately 2.0%, approximately 2.2%, approximately 2.5%, approximately 2.7%, approximately 3.0 %, about 3.2%, about 3.5%, about 3.7%, or about 4.0%.

[00152] As used herein, the term "unconjugated linker" refers to an antibody that is covalently linked to a linker derived from a crosslinking reagent (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1), where the antibody is not covalently linked to the drug (e.g., DM1, DM3, or DM4) via a linker (i.e., "non-conjugated linker" can be represented as Ab-MCC, Ab-SPDB, or Ab-CX1-1).

1. Fragment representing a drug

[00153] The present invention provides immunoconjugates that specifically bind to CCR7. The antibody-drug conjugates of the present invention comprise anti-CCR7 antibodies, antibody fragments (e.g., antigen-binding fragments), or functional equivalents that are conjugated to a drug moiety, e.g., an anticancer agent, an autoimmune drug, an anti-inflammatory agent, an antifungal agent. , an antibacterial agent, an antiparasitic agent, an antiviral agent, or an anesthetic agent. Antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention may be conjugated to multiple identical or different drug fragments using any of the methods known in the art.

[00154] In some embodiments, the drug fragment of the immunoconjugates of the present invention is selected from the group consisting of a V-ATPase inhibitor, a proapoptotic agent, a Bcl2 inhibitor, an MCL1 inhibitor, an HSP90 inhibitor, an IAP inhibitor, an mTor inhibitor, a stabilizer microtubules, microtubule destabilizer, RNA polymerase inhibitor, amanitin, pyrrolobenzodiazepine, auristatin, dolastatin, maytansinoid, MetAP (methionine aminopeptidase), CRM1 nuclear export inhibitor, DPPIV inhibitor, Eg5 inhibitor, proteasome inhibitors, mitochondrial phosphoryl transfer reactions inhibitor, protein synthesis inhibitor , a kinase inhibitor, a CDK2 inhibitor, a CDK9 inhibitor, a kinesin inhibitor, an HDAC inhibitor, a DNA damaging agent, a DNA alkylating agent, a DNA intercalator, a DNA minor groove binding agent, and a DHFR inhibitor.

[00155] In a specific embodiment, the drug moiety of the immunoconjugates of the present invention is auristatin disclosed in PCT Publications WO 2015/095301 and WO2015/189791, both applications hereby incorporated by reference. Non-limiting examples of constructs from an auristatin drug moiety and a linker are:

(which is AURIX2 as disclosed in this application) and

which is AURIX1 as disclosed in this application).

[00156] In one embodiment, the drug moiety of the immunoconjugates of the present invention is a maytansinoid drug moiety, such as, but not limited to, DM1, DM3, or DM4.

[00157] In addition, antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention can be conjugated to a drug moiety that modifies a given biological response. Drug moieties should not be construed as being limited to classic chemical therapeutic agents. For example, a drug moiety may be a protein, peptide, or polypeptide having the desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, Pseudomonas exotoxin, cholera toxin, or diphtheria toxin; a protein such as tumor necrosis factor, α-interferon, β-interferon, nerve growth factor, platelet growth factor, tissue plasminogen activator, cytokine, apoptotic agent, anti-angiogenic agent or biological response modifier such as, for example, lymphokine.

[00158] In one embodiment, the antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention are conjugated to a drug moiety, such as a cytotoxin, a drug (eg, an immunosuppressant), or a radiotoxin. Examples of cytotoxins include, without limitation, taxanes (see, e.g., International (PCT) Patent Application Nos. WO 01/38318 and PCT/US03/02675), DNA alkylating agents (e.g., CC-1065 analogs), anthracyclines, tubulisin analogs , duocarmycin analogs, auristatin E, auristatin F, maytansinoids, and cytotoxic agents containing a reactive polyethylene glycol moiety (see, for example, Sasse et al., J. Antibiot. (Tokyo), 53, 879-85 (2000), Suzawa et al., Bioorg Med. Chem., 8, 2175-84 (2000), Ichimura et al., J. Antibiot (Tokyo), 44, 1045-53 (1991), Francisco et al., Blood ( 2003) (electronic publication prior to printed publication), U.S. Patent Nos. 5475092, 6340701, 6372738 and 6436931, U.S. Patent Application Publication No. 2001/0036923 A1, pending U.S. Patent Applications Serial Nos. 10/024290 and 10/ 116053, and International (PCT) Patent Application No. WO 01/49698), taxon, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenopo zid, vincristine, vinblastine, t. colchicine, doxorubicin, daunorubicin, dihydroxyanthracindione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol and puromycin, as well as their analogues or homologues. Therapeutic agents also include, for example, antimetabolites (eg, methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil, decarbazine), disruptive agents (eg, mechlorethamine, thiotepa, chlorambucil, melphalan, carmustine (BSNU), and lomustine ( CCNU), cyclophosphamide, busulfan, dibromomannite, streptozotocin, mitomycin C and cis-dichlorodiaminplatinum(II) (DDP), cisplatin, anthracyclines (eg, daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (eg, dactinomycin (formerly actinomycin), bleomycin , mithramycin and anthramycin (AMC)) and antimitotic agents (eg vincristine and vinblastine) (See eg US20090304721 from Seattle Genetics).

[00159] Other examples of cytotoxins that can be conjugated to antibodies, antibody fragments (antigen-binding fragments), or functional equivalents of the present invention include duocarmycins, calicheamicins, maytansins, and auristatins, as well as derivatives thereof.

[00160] Various types of cytotoxins, linkers, and methods for conjugating therapeutics to antibodies are known in the art, see, for example, Saito et al., (2003) Adv. drug deliv. Rev. 55:199-215; Trail et al., (2003) Cancer Immunol. Immunother. 52:328-337; Payne, (2003) Cancer Cell 3:207-212; Allen, (2002) Nat. Rev. Cancer 2:750-763; Pastan and Kreitman, (2002) Curr. Opin. Investig. Drugs 3:1089-1091; Senter and Springer, (2001) Adv. drug deliv. Rev. 53:247-264.

[00161] Antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention can also be conjugated with a radioactive isotope to create cytotoxic radiopharmaceuticals called radioimmunoconjugates. Examples of radioactive isotopes that can be conjugated to antibodies for diagnostic or therapeutic use include, but are not limited to, iodine-131, indium-111, yttrium-90, and lutetium-177. Methods for obtaining radioimmunoconjugates are defined in the prior art. Examples of radioimmunoconjugates are commercially available, including Zevalin ^TM (DEC Pharmaceuticals) and Bexxar ^TM (Corixa Pharmaceuticals), and similar methods can be used to obtain radioimmunoconjugates using the antibodies of the present invention. In some embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), which can be attached to the antibody via a linker molecule. Such linker molecules are well known in the art and are described in Denardo et al., (1998) Clin Cancer Res. 4(10):2483-90; Peterson et al., (1999) Bioconjug. Chem. 10(4):553-7; and Zimmerman et al., (1999) Nucl. Med. Biol. 26(8):943-50, each of which is incorporated by reference in its entirety.

[00162] Antibodies, antibody fragments (e.g., antigen-binding fragments), or functional equivalents of the present invention can also be conjugated to a heterologous protein or polypeptide (or fragment thereof, preferably a polypeptide of at least 10, at least 20, at least 30 , at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 amino acids) to create fusion proteins. In particular, the present invention provides fusion proteins containing an antibody fragment (e.g., antigen-binding fragment) described herein (e.g., Fab fragment, Fd fragment, Fv fragment, F(ab) ₂ fragment, VH domain, VH CDR, VL domain, or VL CDR), and a heterologous protein, polypeptide, or peptide.

[00163] Additional fusion proteins can be generated by gene shuffling, motif shuffling, exon shuffling, and/or codon shuffling (collectively referred to as "DNA shuffling") techniques. DNA shuffling can be used to alter the activity of antibodies of the present invention or fragments thereof (eg, producing antibodies or fragments thereof with higher affinity levels and lower dissociation rates). See, in general, US Pat. Nos. 5,605,793; 5,811,238; 5,830,721; Patten et al., (1997) Curr. Opinion Biotechnol. 8:724-33; Harayama, (1998) Trends Biotechnol. 16(2):76-82; Hansson et al., (1999) J. Mol. Biol. 287:265-76; and Lorenzo and Blasco, (1998) Biotechniques 24(2):308-313 (each of these patents and publications is hereby incorporated by reference in its entirety). Antibodies or fragments thereof, or encoded antibodies or fragments thereof, can be modified by subjecting them to random mutagenesis by means of PCR with the introduction of errors, random insertion of nucleotides or other methods prior to recombination. A polynucleotide encoding an antibody or fragment thereof that specifically binds to an antigen may be recombined with one or more components, motifs, sections, moieties, domains, fragments, etc. of one or more heterologous molecules.

[00164] Moreover, antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention can be conjugated to marker sequences, such as a peptide, to facilitate purification. In preferred embodiments, the marker amino acid sequence is a hexahistidine peptide (SEQ ID NO: 628), such as the tag provided in the pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, CA 91311), among others, many of which are commercially available. As described in Gentz et al., (1989) Proc. Natl. Acad. sci. USA 86:821-824, for example, hexahistidine (SEQ ID NO: 628) provides a convenient purification of the fusion protein. Other peptide tags useful for purification include, but are not limited to, a hemagglutinin ("HA") tag that corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., (1984) Cell 37:767) and the "FLAG" tag ( A. Einhauer et al., J. Biochem. Biophys. Methods 49: 455-465, 2001). In accordance with the present invention, antibodies or antigen-binding fragments can also be conjugated to tumor-penetrating peptides to increase the effectiveness of antibodies or antigen-binding fragments.

[00165] In other embodiments, antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention are conjugated to a diagnostic or detectable agent. Such immunoconjugates may be useful in monitoring or predicting the manifestation, development, progression and/or severity of a disease or disorder in a clinical testing procedure such as determining the efficacy of a particular therapy. Such diagnosis and detection can be accomplished by coupling the antibody to detectable substances, including, but not limited to, various enzymes such as, but not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; prosthetic groups such as, but not limited to, streptavidin/biotin and avidin/biotin; fluorescent materials such as without limitation Alexa Fluor 350, Alexa Fluor 405, Alexa Fluor 430, Alexa Fluor 488, Alexa Fluor 500, Alexa Fluor 514, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 555, Alexa Fluor 568, Alexa Fluor 594 , Alexa Fluor 610, Alexa Fluor 633, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor 680, Alexa Fluor 700, Alexa Fluor 750, umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin; luminescent materials such as, but not limited to, luminol; bioluminescent materials such as, but not limited to, luciferase, luciferin, and aequorin; radioactive materials such as, without limitation, iodine ( ¹³¹ I, ¹²⁵ I, ¹²³ I and ¹²¹ I), carbon ( ¹⁴ C), sulfur ( ³⁵ S), tritium ( ³ H), indium ( ¹¹⁵ In, ¹¹³ In, ¹¹² In and ¹¹¹ In), technetium ( ⁹⁹ Tc), thallium ( ²⁰¹ Ti), gallium ( ⁶⁸ Ga, ⁶⁷ Ga), palladium ( ¹⁰³ Pd), molybdenum ( ⁹⁹ Mo), xenon ( ¹³³ Xe), fluorine ( ¹⁸ F), ¹⁵³ Sm, ¹⁷⁷ Lu, ¹⁵⁹ Gd, ¹⁴⁹ Pm, ¹⁴⁰ La, ¹⁷⁵ Yb, ¹⁶⁶ Ho, ⁹⁰ Y, ⁴⁷ Sc, ¹⁸⁶ Re, ¹⁸⁸ Re, ¹⁴² Pr, ¹⁰⁵ Rh, ⁹⁷ Ru, ⁶⁸ Ge, ⁵⁷ Co, ⁶⁵ Zn , ⁸⁵ Sr, ³² P, ¹⁵³ Gd, ¹⁶⁹ Yb, ⁵¹ Cr, ⁵⁴ Mn, ⁷⁵ Se, ⁶⁴ Cu, ¹¹³ Sn and ¹¹⁷ Sn; and positron-active metals using various types of positron emission tomography, and non-radioactive ions of paramagnetic metals.

[00166] Antibodies, antibody fragments (eg, antigen-binding fragments), or functional equivalents of the present invention can also be attached to solid supports, which are particularly useful for immunoassays or target antigen purification. Such solid substrates include, without limitation, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene.

2. Linker

[00167] As used herein, a "linker" is any chemical moiety that is capable of linking an antibody, antibody fragment (eg, antigen-binding fragments), or functional equivalent to another moiety, such as a drug fragment. Linkers may be amenable to cleavage (cleavable linker), such as acid-induced cleavage, photoinduced cleavage, peptidase-induced cleavage, esterase-induced cleavage, glycosidase-induced cleavage, phosphodiesterase-induced cleavage, phosphatase-induced cleavage, and disulfide bond cleavage, under conditions in which the compound or antibody remain active. Alternatively, the linkers may be substantially resistant to cleavage (eg, a stable linker or a non-cleavable linker). In some aspects, the linker is a pre-charged linker, a hydrophilic linker, or a dicarboxylic acid linker.

[00168] In one aspect, the linker used in the present invention is derived from a crosslinking reagent such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl-4-(2-pyridyldithio)pentanoate (SPP ), N-succinimidyl-4-(2-pyridyldithio)butanoate (SPDB), N -succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB), N-succinimidyl iodoacetate (SIA), N- succinimidyl-(4-iodoacetyl)aminobenzoate (SIAB), maleimide-PEG-NHS, N-succinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-sulfosuccinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (sulfo-SMCC) or 2, 5-dioxopyrrolidin-1-yl-17-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5,8,11,14-tetraoxo-4,7,10,13- tetraazaheptadecan-1-oate (CX1-1).

[00169] Non-cleavable linkers are any chemical moiety that is capable of linking a drug, such as a maytansinoid, to an antibody via a stable covalent bond and does not fall into the categories listed above for cleavable linkers. Thus, non-cleavable linkers are largely resistant to acid-induced cleavage, photo-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage. In addition, non-cleavable refers to the ability of a chemical bond in or attached to a linker to resist cleavage induced by an acid, a photolabile cleaving agent, a peptidase, an esterase, or a chemical or physiological compound that cleaves a disulfide bond under conditions where a drug such as a maytansinoid or antibody does not lose its activity.

[00170] Acid labile linkers are linkers that are cleavable at acidic pH. For example, some intracellular compartments, such as endosomes and lysosomes, have an acidic pH (pH 4-5) and provide conditions suitable for cleavage of acid-labile linkers.

[00171] Photolabile linkers are linkers that are applicable on the surface of the body and in many body cavities that are exposed to light. In addition, infrared light can penetrate into the tissue.

[00172] Some linkers can be cleaved by peptidases, ie. are peptidase-cleavable linkers. Only some peptides are easily cleaved inside or outside of cells, see, for example, Trout et al., 79 Proc. Natl. Acad. sci. USA, 626-629 (1982) and Umemoto et al. 43 Int. J. Cancer, 677-684 (1989). In addition, peptides are composed of α-amino acids and peptide bonds, which are chemically amide bonds between the carboxyl group of one amino acid and the amino group of a second amino acid. It is understood that other amide bonds, such as the bond between the carboxyl group and the ε-amino group of lysine, are not peptide bonds and are considered non-cleavable.

[00173] Some linkers can be cleaved by esterases, ie. are esterase-cleavable linkers. Again, only some esters can be cleaved by esterases present inside or outside the cells. Esters are formed by the condensation of a carboxylic acid and an alcohol. Ethers are esters made with simple alcohols such as aliphatic alcohols and small cyclic and small aromatic alcohols.

[00174] Pre-charged linkers are derived from charged cross-linking reagents that retain their charge after being introduced into the antibody-drug conjugate. Examples of pre-charged linkers can be found in US 2009/0274713.

3. Conjugation and preparation of ADC

[00175] Numerous methods for conjugating linker payloads to an antigen-binding moiety are known in the art (for a review see, for example, Antibody-Drug Conjugate, Methods in Molecular Biology, Vol. 1045, Editor L. Ducry, Humana Press (2013)) . Traditionally, drugs are conjugated to native lysine or native cysteine residues of the antibody. The preparations obtained are complex mixtures. Recently, site-specific conjugation techniques have been used to improve the therapeutic index and homogeneity of ADC preparations (reviewed in: Panowski, S.; Bhakta, S.; Raab, H.; Polakis, P.; Junutula, JR mAbs 2014, 6 . 34). Beyond the application of glycoengineering (Zhou, Q. et al. Bioconjugate chemistry 2014, 25 , 510; Zhu, Z. et al. mAbs 2014, 6 , 1190); some of the more well-known methods for producing site-specific ADCs are based on the introduction of engineered cysteines (Junutula, JR et al., Nature biotechnology 2008, 26 , 925; Shinmi, D. et al., Bioconjugate chemistry 2016, 27 , 1324), non-canonical amino acids ( Tian, F. et al., Proceedings National Academy of Sciences USA 2014, 111 , 1766; Axup, JY et al., Proceedings National Academy of Sciences USA 2012, 109 , 16101) or short peptide sequences in an antibody backbone (Drake, PM et al., Bioconjugate chemistry 2014, 25 , 1331; Strop, P. et al., Chemistry & biology 2013, 20 , 161; Beerli, RR et al., PloS one 2015, 10 , e0131177; Grunewald, J. et al. ., Bioconjugate chemistry 2015, 26 , 2554). These methods provide control over cytotoxin stoichiometry and site of attachment, resulting in better pharmacokinetics (PK), safety and efficacy profiles of conjugates compared to conventionally prepared ADCs.

[00176] The conjugates of the present invention can be prepared using any of the methods known in the art, such as those described in US Pat. /0253021 and 2012/0259100 and PCT Publications WO2014/124316 and WO2015/138615. A full summary of the ideas behind these patents and patent application publications is incorporated herein by reference.

Conjugation method with engineered antibody cysteine residues

[00177] the Conjugates of the present invention can be obtained using cysteine residues introduced by design in the antibody, for example, using site-directed mutagenesis. Such site-specific conjugates are homogeneous and have improved properties (Junutula JR, Raab H, Clark S, Bhakta S, Leipold DD, Weir S, Chen Y, Simpson M, Tsai SP, Dennis MS, Lu Y, Meng YG, Ng C , Yang J, Lee CC, Duenas E, Gorrell J, Katta V, Kim A, McDorman K, Flagella K, Venook R, Ross S, Spencer SD, Lee Wong W, Lowman HB, Vandlen R, Sliwkowski MX, Scheller RH, Polakis P, Mallet W. (2008) Nature Biotechnology 26:925-932.)

[00178] Because engineered cysteines in antibodies expressed in mammalian cells are modified during biosynthesis by adducts (disulfides) such as glutathione (GSH) and/or cysteine (Chen et al . 2009) originally expressed by the engineered cysteine residues in the product do not react with reagents that react with thiols, such as maleimide or bromine or iodo-acetamide groups. In order to effect conjugation of the payload to the engineered cysteine after expression, the glutathione or cysteine adducts must be removed by reduction of these disulfide adducts, which usually also causes the restoration of native disulfides in the expressed protein. Removal of adducts from engineered cysteines can be accomplished by first exposing the antibody to a reducing agent, such as dithiothreitol (DTT), TCEP, or reduced cysteine, followed by a procedure that allows all native disulfide bonds of the antibody to be reoxidized to reduce and/or stabilize functional structure of the antibody.

[00179] Several methods of reducing and reoxidizing antibodies with engineered Cys residues can be used to produce antibody-drug conjugates. Attempts to follow reoxidation protocols previously described in the literature using high concentrations of CuSO ₄ resulted in protein precipitation (Junutula JR, Raab H, Clark S, Bhakta S, Leipold DD, Weir S, Chen Y, Simpson M, Tsai SP, Dennis MS, Lu Y, Meng YG, Ng C, Yang J, Lee CC, Duenas E, Gorrell J, Katta V, Kim A, McDorman K, Flagella K, Venook R, Ross S, Spencer SD, Lee Wong W, Lowman HB, Vandlen R, Sliwkowski MX, Scheller RH, Polakis P, Mallet W. (2008) Nature Biotechnology 26:925). The present inventors have successfully prepared and prepared antibody-drug conjugates using several different antibody reduction and reoxidation techniques.

[00180] In one example, freshly generated DTT is added to purified Cys mutated antibodies to a final concentration of 10 mM. After incubation with DTT at room temperature for 1 hour, the mixture was dialyzed against PBS at 4° C. for three days with daily buffer changes to remove DTT and reoxidize the native disulfide bonds of the antibody. An alternative method is to remove the reducing reagents by passing through a desalting column such as Sephadex G-25 equilibrated with PBS. After the protein has completely recovered, 1 mM oxidized ascorbate (dihydroascorbic acid) is optionally added to the desalted samples and incubations for re-oxidation are carried out for 20-24 hours.

[00181] In another illustrative method, release of engineered Cys residues is accomplished by adding fully reduced cysteine at a concentration of 20 mM to antibodies bound to Protein A Sepharose resin. minutes at room temperature, then the reducing agent is quickly removed by washing the resin with 50 bed volumes of PBS. Re-oxidation of the reduced antibody is achieved by incubating the washed suspension at room temperature with or without the addition of 50-2000 nM CuCl ₂ as a catalyst. With the exception of the use of copper sulfate, the examples of this document used each of the protocols described in this document, with similar results. Reoxidation reduces the intrachain disulfides, while dialysis, desalting, or protein A chromatography removes the reducing agent, as well as the cysteines and glutathiones originally attached to the antibody cysteine(s) introduced by construction. Reverse phase HPLC is commonly used to monitor the reoxidation process: antibodies are loaded onto a PLRP-S column (4000 Å, 50 mm × 2.1 mm, Agilent) heated to 80°C and eluted using a linear gradient of 30-45 % CH ₃ CN in water containing 0.1% TFA at 1.5 ml/min and detection peak at 215, 254 and 280 nm.

[00182] After re-oxidation, the antibody is conjugated to a preformed linker-drug fragment. As an example, a preformed linker drug fragment (such as, for example, MMTBT-DM4; MPET-DM4; MBT-DM4; MEPET-DM4, MPBT-DM1; and other linker drug fragments, described herein) are added to the re-oxidized Cys-mutated antibody at 10 molar equivalents relative to the antibody in PBS buffer (pH 7.2). Incubations are carried out for 1 hour. The conjugation process is monitored by reverse phase HPLC, which can separate conjugated antibodies from non-conjugated ones. Reaction mixtures during conjugation are analyzed on a PRLP-S column (4000 Å, 50 mm × 2.1 mm, Agilent) heated to 80°C, and the column is eluted with a linear gradient of 30-60% acetonitrile in water containing 0 .1% TFA at a flow rate of 1.5 ml/min. The elution of proteins from the column is monitored at 280 nm, 254 nm and 215 nm.

[00183] In one embodiment, examples of a linker drug moiety for cysteine conjugation can be prepared according to Schemes 1-3:

Scheme 1.

where:

the drug moiety is attached to the linker via a thiol functional group;

L ¹ represents C _1-6 alkylene, while one of the methylene groups can be replaced by oxygen;

L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; and

X is -C(O)-NH-, -NHC(O)- or triazole;

where alkylene is linear or branched; and

RG1 and RG2 are 2 reactive groups forming the X group.

The reaction of groups that form an amide or triazole is well known in the art.

One example of the preformation of a linker drug moiety is shown in Scheme 2, where the drug moiety is DM4; RG1 is an amino group and RG2 is an activated acid, which lead to the formation of an amide bond (X):

Scheme 2.

Another example of pre-formation of a linker-drug moiety is shown in Scheme 3, where the drug moiety is DM4; RG1 is an azide group and RG2 is an alkyne group, which lead to the formation of a tetrazole (X):

Scheme 3.

[00184] The efficiency of conjugation of various maleimide-attached drug moieties to a Cys-mutated antibody varies depending on the solubility of the drug moiety used, however, many reactions result in over 90% conjugate. To evaluate the state of aggregation, the resulting conjugates are analyzed on a size exclusion chromatography column (GE, Superdex200, 3.2/30) at a flow rate of 0.1 ml/min in PBS. All conjugates are predominantly monomeric. Most of the conjugates contain less than 3% dimeric and oligomeric material, indicating that conjugation of the maleimide-attached drug moiety to the Cys-mutated antibody does not cause aggregation.

[00185] Immunoconjugates are also characterized in terms of the average load of drug moiety per antibody binding moiety, commonly referred to as the drug-to-antibody ratio (DAR). The DAR value is extrapolated from, for example, LC-MS data for reconstituted and deglycosylated samples. LC/MS provides the ability to quantify the average number of payload molecules (drug-forming fragment) attached to an antibody in an ADC. HPLC separates the antibody into light and heavy chains, and also separates the heavy chain (HC) and light chain (LC) according to the number of linker-payload groups per chain. The mass spectrum data allows identification of the species in the mixture, e.g. LC, LC+1, LC+2, HC, HC+1, HC+2, etc. Based on the average load of the LC and HC circuits for the ADC, an average DAR can be calculated. The DAR for a given immunoconjugate sample is the average number of drug molecules (payload) attached to a tetrameric antibody containing two light chains and two heavy chains.

Method for conjugation with native cysteine residues of an antibody

[00186] The linker drug fragments described herein can also be conjugated to native cysteine residues of unengineered antibodies using a procedure that involves partial antibody recovery (Doronina, SO, Toki, BE, Torgov, MY, Mendelsohn, BA, Cerveny, CG, Chace, DF, DeBlanc, RL, Gearing,RP, Bovee, TD, Siegall, CB, Francisco, JA, Wahl, AF, Meyer, DL, and Senter, PD (2003) Development of potent monoclonal antibody auristatin conjugates for cancer therapy Nat Biotechnol 21 , 778-784). The following protocol is a non-limiting example of how such conjugates can be made. Interchain and intrachain disulfide bonds of the antibody (typically at a concentration of 5-10 mg/ml) are first partially reduced in PBS containing 2 mM EDTA by adding TCEP to a final concentration of 10 mM and incubating the mixture at 37°C for 1 hour. After desalting and adding 1% w/v PS-20 detergent, partially reduced antibodies (1-2 mg/mL) are reacted overnight at 4°C with 0.5-1 mg of a maleimide-containing linker compound and payload per 10 mg of antibody . The resulting conjugates are purified by protein A chromatography using standard methods and buffer exchanged with PBS, and are typically analyzed by mass spectrometry (MS), analytical size exclusion chromatography (AnSEC) and analytical hydrophobic interaction chromatography (AnHIC) to determine the ratio of drug- k-antibody, aggregation propensity and hydrophobicity, as well as using activity assays.

One-step method for cross-linking with lysine residues of an antibody

[00187] In one embodiment, the conjugates of the present invention can be obtained using a one-step method of cross-linking a drug with lysine residues on an antibody. The method involves combining the antibody, drug, and crosslinker in a substantially aqueous medium, optionally containing one or more co-solvents, at a suitable pH. In one embodiment, the method comprises the step of contacting an antibody of the present invention with a drug (e.g., DM1 or DM4) to form a first antibody/drug mixture, and then contacting the first antibody/drug mixture with a crosslinker (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) in a solution having a pH of about 4 to about 9 to form a mixture containing (i) a conjugate (e.g., Ab-MCC-DM1, Ab-SPDB-DM4 or Ab-CX1-1-DM1), (ii) free drug (e.g., DM1 or DM4) and (iii) reaction by-products.

[00188] In one embodiment, the one-step method involves bringing the antibody into contact with a drug (e.g., DM1 or DM4) and then with a crosslinker (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) in a solution having a pH of about 6 or greater (e.g., about 6 to about 9, about 6 to about 7, about 7 to about 9, about 7 to about 8.5, about 7.5 to about 8.5, about 7.5 to about 8, 0, about 8.0 to about 9.0, or about 8.5 to about 9.0). For example, the method of the present invention involves contacting a cell-binding agent with a drug (DM1 or DM4) and then with a cross-linking agent (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) in a solution having a pH of about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5 , about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5 , about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, about 8.5 , about 8.6, about 8.7, about 8.8, about 8.9, or about 9.0. In a specific embodiment, the method of the present invention involves bringing a cell-binding agent into contact with a drug (e.g., DM1 or DM4) and then with a crosslinker (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) in a solution having a pH of approximately 7.8 (e.g., pH 7.6 to 8.0 or pH 7.7 to 7.9).

[00189] One-step method (i.e., bringing the antibody into contact with a drug (for example, DM1 or DM4) and then with a crosslinker (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1) can be carried out at any suitable temperature known in the art. For example, a one-step process can take place at about 20°C or less (for example, from approximately -10°C (provided that the solution is not allowed to freeze, for example, due to the presence of an organic solvent used to dissolve the cytotoxic agent and the bifunctional crosslinker) to about 20°C, from about 0°C to about 18°C, from about 4°C to about 16°C), at room temperature (for example , from about 20°C to about 30°C or from about 20°C to about 25°C) or at an elevated temperature (for example, from about 30°C to about 37°C). In one embodiment, the one-step process occurs at a temperature of from about 16°C to about 24°C (e.g., about 16°C, about 17°C, about 18°C, about 19°C, about 20°C, about 21°C C, approximately 22°C, approximately 23°C, approximately 24°C or approximately 25°C). In another embodiment, the one-step process is carried out at a temperature of about 15°C or less (eg, from about -10°C to about 15°C or from about 0°C to about 15°C). For example, the method involves bringing the antibody into contact with a drug (e.g., DM1 or DM4) and then with a crosslinker (e.g. SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1) at approximately 15°C, approximately 14°C, approximately 13°C, approximately 12°C , approximately 11°C, approximately 10°C, approximately 9°C, approximately 8°C, approximately 7°C, approximately 6°C, approximately 5°C, approximately 4°C, approximately 3°C, approximately 2°C , approximately 1°C, approximately 0°C, approximately -1°C, approximately -2°C, approximately -3°C, approximately -4°C, approximately -5°C, approximately -6°C, approximately -7 °C, approximately -8°C, approximately -9°C or approximately -10°C, provided that the solution is not allowed to freeze, for example, due to the presence of the organic solvent(s) used ) to dissolve the crosslinker (eg SMCC, sulfo-SMCC, sulfo-SPDB, SPDB or CX1-1). In one embodiment, the method involves contacting the antibody with a drug (eg, DM1 or DM4) and then with a crosslinker (eg, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) at a temperature of from about - 10°C to approximately 15°C, approximately 0°C to approximately 15°C, approximately 0°C to approximately 10°C, approximately 0°C to approximately 5°C, approximately 5°C to approximately 15 °C, from about 10°C to about 15°C, or from about 5°C to about 10°C. In another embodiment, the method involves contacting the antibody with a drug (eg, DM1 or DM4) and then with a crosslinker (eg, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) at a temperature of approximately 10°C. C (e.g. 8°C to 12°C or 9°C to 11°C).

[00190] In one embodiment, the contacting described above is carried out by introducing an antibody, then contacting the antibody with a drug (e.g., DM1 or DM4) to form a first mixture containing the antibody and the drug (e.g., DM1 or DM4), and then bringing the first mixture containing the antibody and drug (eg DM1 or DM4) into contact with a crosslinker (eg SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1). For example, in one embodiment, an antibody is added to a reaction vessel, a drug (e.g., DM1 or DM4) is added to the reaction vessel (thereby contacting the antibody), and then a crosslinker (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1) is added to the mixture containing the antibody and drug (eg, DM1 or DM4) (thereby bringing into contact with the mixture containing the antibody and drug). In one embodiment, the antibody is added to the reaction vessel and the drug (eg, DM1 or DM4) is added to the reaction vessel immediately after the antibody is added to the vessel. In another embodiment, the antibody is added to the reaction vessel and the drug (e.g., DM1 or DM4) is added to the reaction vessel some time after the antibody is added to the vessel (e.g., about 5 minutes, about 10 minutes, about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 1 hour, about 1 day or more after the cell-binding agent is added to the container). The drug (eg, DM1 or DM4) can be added rapidly (ie, in a short time frame such as about 5 minutes, about 10 minutes) or slowly (such as using a pump).

[00191] A mixture containing an antibody and a drug (e.g., DM1 or DM4) can then be contacted with a crosslinker (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1), either immediately after exposure of the antibody to the drug (eg, DM1 or DM4) or slightly later (eg, from about 5 minutes to about 8 hours or more) after bringing the antibody into contact with the drug (for example, DM1 or DM4). For example, in one embodiment, a cross-linking agent (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) is added to the mixture containing the antibody and drug (e.g., DM1 or DM4), immediately after the addition of the drug (for example, DM1 or DM4) into the reaction vessel containing the antibody. Alternatively, a mixture containing antibody and drug (eg, DM1 or DM4) may be contacted with a crosslinker (eg, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) after about 5 minutes, approximately 10 minutes, approximately 20 minutes, approximately 30 minutes, approximately 1 hour, approximately 2 hours, approximately 3 hours, approximately 4 hours, approximately 5 hours, approximately 6 hours, approximately 7 hours, approximately 8 hours or more after the antibody has been contacted with medicine (for example, DM1 or DM4).

[00192] After the mixture containing the antibody and drug (for example, DM1 or DM4) are brought into contact with a cross-linking agent (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) allow the reaction to proceed for about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours , approximately 8 hours, approximately 9 hours, approximately 10 hours, approximately 11 hours, approximately 12 hours, approximately 13 hours, approximately 14 hours, approximately 15 hours, approximately 16 hours, approximately 17 hours, approximately 18 hours, approximately 19 hours, approximately 20 hours, approximately 21 hours, approximately 22 hours, approximately 23 hours, approximately 24 hours or more (for example, approximately 30 hours, approximately 35 hours, approximately 40 hours, approximately 45 hours, or approximately 48 hours).

[00193] In one embodiment, the one-step method further comprises a blocking step to block any unreacted drug (e.g., DM1 or DM4) and/or unreacted crosslinker (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1- one). Typically, the blocking step is carried out prior to purification of the conjugate. In one embodiment, the mixture is blocked by bringing the mixture into contact with a blocking reagent. As used herein, "blocking reagent" refers to a reagent that reacts with free drug (e.g., DM1 or DM4) and/or cross-linking agent (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1). In one embodiment, maleimide or haloacetamide blocking reagents, such as 4-maleimidobutyric acid, 3-maleimidopropionic acid, N-ethylmaleimide, iodoacetamide, or iodoacetamidopropionic acid, can be used to ensure that any unreacted group (such as a thiol) in the drug (e.g. , DM1 or DM4) is locked. The blocking step may help prevent dimerization of the drug (eg, DM1). Dimerized DM1 can be difficult to remove. After blocking with polar, charged, thiol-blocking reagents (such as 4-maleimidobutyric acid or 3-maleimidopropionic acid), excess, unreacted DM1 is converted to a polar, charged, water-soluble adduct that can be easily separated from the covalently bonded conjugate during purification steps. Blocking with nonpolar and neutral thiol blocking reagents can also be used. In one embodiment, the mixture is blocked by contacting the mixture with a blocking agent that reacts with unreacted crosslinker (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1). For example, nucleophiles can be added to the mixture to block any unreacted SMCC. Preferably, the nucleophile is an amino group-containing nucleophile such as lysine, taurine and hydroxylamine.

[00194] In a preferred embodiment, the reaction is allowed to proceed (i.e., the antibody is brought into contact with the drug (e.g., DM1 or DM4) and then with a crosslinker (eg SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1)) to completion before contacting the mixture with the blocking agent. Therefore, the blocking agent is added to the mixture after about 1 hour to about 48 hours (for example, after about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, approximately 14 hours, approximately 15 hours, approximately 16 hours, approximately 17 hours, approximately 18 hours, approximately 19 hours, approximately 20 hours, approximately 21 hours, approximately 22 hours, approximately 23 hours, approximately 24 hours or from approximately 25 hours to approximately 48 hours) after the admixture containing the antibody and the drug (for example, DM1 or DM4), in contact with a crosslinker (for example, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1).

[00195] Alternatively, the mixture is blocked by lowering the pH of the mixture to about 5.0 (e.g., 4.8, 4.9, 5.0, 5.1 or 5.2). In another embodiment, the mixture is blocked by lowering the pH to less than 6.0, less than 5.5, less than 5.0, less than 4.8, less than 4.6, less than 4.4, less than 4.2, less than 4.0. Alternatively, the pH is reduced to a value of about 4.0 (for example, 3.8, 3.9, 4.0, 4.1 or 4.2) to about 6.0 (e.g. 5.8, 5.9, 6.0, 6.1, or 6.2), from about 4.0 to about 5.0, from about 4.5 (e.g., 4.3, 4.4, 4.5, 4.6 or 4.7) to about 5.0. In one embodiment, the mixture is blocked by lowering the pH of the mixture to 4.8. In another embodiment, the mixture is blocked by lowering the pH of the mixture to 5.5.

[00196] In one embodiment, the one-step method further includes a retention step to release unstable bound linkers from the antibody. The retention step involves holding the mixture until the conjugate is purified (eg, after the reaction step, between the reaction step and the blocking step, or after the blocking step). For example, the method involves (a) bringing the antibody into contact with a drug (e.g., DM1, DM3 or DM4) to form a mixture containing the antibody and drug (eg DM1, DM3 or DM4); and then bringing the mixture containing the antibody and drug (eg, DM1, DM3, or DM4) into contact with a crosslinker (eg, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) in a solution having a pH of about 4 to about 9, to obtain a mixture containing (i) a conjugate (for example, Ab-MCC-DM1, Ab-SPDB-DM4 or Ab-CX1-1-DM1), (ii) free drug (for example, DM1, DM3 or DM4) and (iii) by-products of the reaction, (b) holding the mixture obtained in step (a) until the unstable bound linkers are released from the cell-binding agent, and (c) purifying the mixture to obtain a purified conjugate.

[00197] In another embodiment, the method comprises (a) contacting an antibody with a drug (eg, DM1, DM3, or DM4) to form a mixture containing the antibody and the drug (eg, DM1, DM3, or DM4); and then bringing the mixture containing the antibody and drug (eg, DM1, DM3, or DM4) into contact with a crosslinker (eg, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) in a solution having a pH of about 4 to about 9 to form a mixture containing (i) the conjugate, (ii) the free drug (e.g., DM1, DM3 or DM4) and (iii) reaction by-products, (b) blocking the mixture obtained in step (a) to block any unreacted drug (e.g. DM1, DM3 or DM4) and/or unreacted crosslinker (e.g. SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1), (c) holding the mixture obtained in step (b) until the unstable bound linkers are released from the cell-binding agent, and (d) purifying the mixture to obtain a purified conjugate (e.g. , Ab-MCC-DM1, Ab-SPDB-DM4 or Ab-CX1-1-DM1).

[00198] Alternatively, the retention step can be performed after purification of the conjugate, followed by an additional purification step.

[00199] In a preferred embodiment, the reaction is allowed to proceed to completion before the retention step. Therefore, the retention step is performed after about 1 hour to about 48 hours (e.g., after about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, approximately 9 hours, approximately 10 hours, approximately 11 hours, approximately 12 hours, approximately 13 hours, approximately 14 hours, approximately 15 hours, approximately 16 hours, approximately 17 hours, approximately 18 hours, approximately 19 hours, approximately 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours, or from about 24 hours to about 48 hours) after bringing the antibody/drug mixture (e.g., DM1, DM3, or DM4) into contact with the crosslinker ( e.g. SMCC, sulfo-SMCC, SPDB, sulfo-SPDB or CX1-1).

[00200] The retention step involves maintaining the solution at a suitable temperature (e.g., from about 0°C to about 37°C) for a suitable period of time (for example, from about 1 hour to about 1 week, from about 1 hour to about 24 hours, from about 1 hour to about 8 hours, or from about 1 hour up to about 4 hours) to release stably bound linkers from the antibody without significantly releasing stably bound linkers from the antibody. In one embodiment, the retention step involves maintaining the solution at about 20° C. or less (e.g., from about 0°C to about 18°C, from about 4°C to about 16°C), at room temperature (for example, from about 20°C to about 30°C or from about 20°C to about 25°C) or at an elevated temperature (for example, from about 30°C to about 37°C). In one embodiment, the retention step involves maintaining the solution at a temperature of from about 16°C to about 24°C (for example, approximately 15°C, approximately 16°C, approximately 17°C, approximately 18°C, approximately 19°C, approximately 20°C, approximately 21°C, approximately 22°C, approximately 23°C, approximately 24°C or approximately 25°C). In another embodiment, the retention step involves maintaining the solution at a temperature of from about 2°C to about 8°C (for example, approximately 0°C, approximately 1°C, approximately 2°C, approximately 3°C, approximately 4°C, approximately 5°C, approximately 6°C, approximately 7°C, approximately 8°C, approximately 9°C or approximately 10°C). In another embodiment, the retention step involves maintaining the solution at a temperature of about 37°C (e.g., about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40 °C).

[00201] The duration of the retention step depends on the temperature and pH at which the retention step is carried out. For example, the duration of the retention step can be significantly reduced by conducting the retention step at an elevated temperature, with the maximum temperature being limited by the stability of the cell-binding agent-cytotoxic agent conjugate. The retention step may include maintaining the solution for from about 1 hour to about 1 day (e.g., about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours , about 9 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, or about 24 hours), from about 10 hours to about 24 hours, from about 12 hours to about 24 hours, from about 14 hours to about 24 hours, from about 16 hours to about 24 hours, from about 18 hours to about 24 hours, from about 20 hours to about 24 hours, from about 5 hours to about 1 week , from approx. 20 hours to approx. 1 week, from approx. 12 hours to about 1 week (e.g., about 12 hours, about 16 hours, about 20 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, or about 7 days) or from about 1 day to about 1 week.

[00202] In one embodiment, the retention step involves maintaining the solution at a temperature of from about 2°C to about 8°C for a period of at least about 12 hours up to a week. In another embodiment, the retention step involves maintaining the solution at a temperature of from about 2° C. to about 8° C. overnight (eg, from about 12 to about 24 hours, preferably about 20 hours).

[00203] The pH value for the retention step is preferably from about 4 to about 10. In one embodiment, the pH value for the retention step is about 4 or more, but less than about 6 (for example, from 4 to 5.9), or about 5 or more but less than about 6 (eg, 5 to 5.9). In another embodiment, the pH values for the retention step are in the range of about 6 to about 10 (eg, about 6.5 to about 9, about 6 to about 8). For example, pH values for the retention step may be about 6, about 6.5, about 7, about 7.5, about 8, about 8.5, about 9, about 9.5, or about 10.

[00204] In specific embodiments, the retention step may include incubating the mixture at 25°C at a pH of about 6-7.5 for about 12 hours to about 1 week, incubating the mixture at 4°C at a pH of about 4.5-5 ,9 for about 5 hours to about 5 days, or incubating the mixture at 25°C at a pH of about 4.5-5.9 for about 5 hours to about 1 day.

[00205] The one-step method may optionally include the addition of sucrose during the reaction step to increase solubility and recover conjugates. Desirably, sucrose is added at a concentration of from about 0.1% (w/v) to about 20% (w/v) (e.g., about 0.1% (w/v), 1% (w/v), 5% ( w/v), 10% (w/v), 15% (w/v) or 20% (w/v)). Preferably, sucrose is added at a concentration of from about 1% (w/v) to about 10% (w/v) (e.g., about 0.5% (w/v), about 1% (w/v), about 1.5 % (w/v), approx 2% (w/v), approx 3% (w/v), approx 4% (w/v), approx 5% (w/v), approx 6% (w/v) ), about 7% (w/v), about 8% (w/v), about 9% (w/v), about 10% (w/v), or about 11% (w/v)). In addition, the reaction step may also include the addition of a buffering agent. Any suitable buffer means known in the art may be used. Suitable buffering agents include, for example, citrate buffer, acetate buffer, succinate buffer and phosphate buffer. In one embodiment, the buffering agent is selected from the group consisting of HEPPSO (N-(2-hydroxyethyl)piperazine-N'-(2-hydroxypropanesulfonic acid)), POPSO (piperazine-1,4-bis-(2-hydroxypropanesulfonic acid) dehydrate )), HEPES (4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid), HEPPS (EPPS) (4-(2-hydroxyethyl)piperazine-1-propanesulfonic acid), TES (N-[tris(hydroxymethyl)methyl ]-2-aminoethanesulfonic acid) and combinations thereof.

[00206] In one embodiment, the one-step method may further comprise the step of purifying the mixture to obtain a purified conjugate (eg, Ab-MCC-DM1, Ab-SPDB-DM4, or Ab-CX1-1-DM1). Any purification methods known in the art can be used to purify the conjugates of the present invention. In one embodiment, the conjugates of the present invention are purified using tangential flow filtration (TFF), non-adsorption chromatography, adsorption chromatography, adsorption filtration, selective precipitation, or any other suitable purification method, as well as combinations thereof. In another embodiment, prior to subjecting the conjugates to the purification process described above, the conjugates are first filtered through one or more PVDF membranes. Alternatively, the conjugates are filtered through one or more PVDF membranes after subjecting the conjugates to the purification process described above. For example, in one embodiment, the conjugates are filtered through one or more PVDF membranes and then purified using tangential flow filtration. Alternatively, the conjugates are purified using tangential flow filtration and then filtered through one or more PVDF membranes.

[00207] Any suitable TFF system may be used for purification, including the Pellicon type system (Millipore, Billerica, Massachusetts), the Sartocon cassette system (Sartorius AG, Edgewood, New York), and the Centrasette type system (Pall Corp., East Hills, New York).

[00208] Any suitable adsorption chromatography resin can be used for purification. Preferred adsorption chromatography resins include hydroxyapatite chromatography, charge induction hydrophobic chromatography (HCIC), hydrophobic interaction chromatography (HIC), ion exchange chromatography, combined mode ion exchange chromatography, immobilized metal ion affinity chromatography (IMAC), dye ligand chromatography , affinity chromatography, reverse phase chromatography, and combinations thereof. Examples of suitable hydroxyapatite resins include ceramic hydroxyapatite (CHT type I and type II, Bio-Rad Laboratories, Hercules, CA), HA Ultrogel hydroxyapatite (Pall Corp., East Hills, NY) and ceramic fluorapatite (CFT type I and type II, Bio-Rad Laboratories, Hercules, CA). An example of a suitable resin for HCIC is MEP Hypercel resin (Pall Corp., East Hills, New York). Examples of suitable HIC resins include Butyl Sepharose, Hexyl Sepharose, Phenyl Sepharose, and Octyl Sepharose (all from GE Healthcare, Piscataway, NJ), as well as Macro-prep methyl resin and Macro tert-butyl resin. -Prep (Biorad Laboratories, Hercules, CA). Examples of suitable ion exchange resins include SP-Sepharose, CM-Sepharose and Q-Sepharose resins (all from GE Healthcare, Piscataway, NJ) and Unosphere S resin (Bio-Rad Laboratories, Hercules, CA). Examples of suitable resins for combined mode ion exchange chromatography include Bakerbond ABx resin (JT Baker, Phillipsburg, NJ). Examples of suitable IMAC resins include Chelating Sepharose resin (GE Healthcare, Piscataway, NJ) and Profinity IMAC resin (Bio-Rad Laboratories, Hercules, CA). Examples of suitable dye ligand chromatography resins include Blue Sepharose resin (GE Healthcare, Piscataway, NJ) and Affi-gel Blue resin (Bio-Rad Laboratories, Hercules, CA). Examples of suitable affinity chromatography resins include protein A Sepharose resin (e.g., MabSelect, GE Healthcare, Piscataway, NJ) and lectin affinity chromatography resins, e.g., Lentil Lectin Sepharose resin (GE Healthcare, Piscataway). , NJ) where the antibody carries appropriate lectin binding sites. Examples of suitable reverse phase chromatography resins include C4, C8, and C18 resins (Grace Vydac, Hesperia, Calif.).

[00209] Any suitable non-adsorption chromatography resin can be used for purification. Examples of suitable non-adsorption chromatography resins include, but are not limited to, SEPHADEX™ G-25, G-50, G-100, SEPHACRYL™ resins (e.g., S-200 and S-300), SUPERDEX™ resins (e.g., SUPERDEX™ 75 and SUPERDEX™ 200), BIO-GEL® resins (eg P-6, P-10, P-30, P-60 and P-100), and others known to those of ordinary skill in the art.

Two-step and one-pot method for cross-linking with antibody lysine residues

[00210] In one embodiment, the conjugates of the present invention can be prepared as described in US Patent No. 7,811,572 and US Patent Application Publication No. 2006/0182750. The method includes the steps of: (a) contacting an antibody of the present invention with a crosslinker (e.g., SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1) with a covalently attached linker (i.e., Ab- SMCC, Ab-SPDB or Ab-CX1-1) to the antibody and thereby obtain a first mixture containing the antibody having a linker associated with it; (b) optionally subjecting the first mixture to a purification method to obtain a purified first mixture from an antibody having a linker associated therewith; (c) conjugating the drug (e.g., DM1, DM3, or DM4) to an antibody having an associated linker in the first mixture by reacting the antibody having an associated linker with the drug (e.g., DM1, DM3, or DM4) in a solution having a pH of about 4 to about 9 to form a second mixture containing (i) a conjugate (e.g., Ab-MCC-DM1, Ab-SPDB-DM4, or Ab-CX1-1-DM1), (ii) free drug (eg DM1, DM3 or DM4) and (iii) reaction intermediates; and (d) subjecting the second mixture to a purification process to purify the conjugate from the other components of the second mixture. Alternatively, purification step (b) may be omitted. In steps (b) and (d), any of the purification methods described herein can be used. In one embodiment, both steps (b) and (d) use TFF. In another embodiment, TFF is used in step (b) and adsorption chromatography (eg, CHT) is used in step (d).

Reagent for one-step method and in-situ method for cross-linking with lysine residues of an antibody

[00211] In one embodiment, the conjugates of the present invention can be prepared by conjugating a preformed linker-drug compound (e.g., SMCC-DM1, sulfo-SMCC-DM1, SPDB-DM4, or CX1-1-DM1) with an antibody of the present invention. of the invention as described in US Patent No. 6,441,163 and US Patent Application Publication Nos. 2011/0003969 and 2008/0145374, followed by a purification step. You can use any of the cleaning methods described in this document. A linker-drug compound is prepared by reacting a drug (eg, DM1, DM3, or DM4) with a cross-linking agent (eg, SMCC, sulfo-SMCC, SPDB, sulfo-SPDB, or CX1-1). The linker-drug compound (eg, SMCC-DM1, sulfo-SMCC-DM1, SPDB-DM4, or CX1-1-DM1) is optionally purified prior to antibody conjugation.

Antibodies to CCR7

[00212] The present invention provides antibodies or antibody fragments (eg, antigen-binding fragments) that specifically bind to CCR7. The antibodies or antibody fragments (eg, antigen-binding fragments) of the present invention include, without limitation, human monoclonal antibodies or fragments thereof isolated as described in the examples.

[00213] In some embodiments, the present invention provides antibodies or antibody fragments (e.g., antigen-binding fragments) that specifically bind CCR7, wherein said antibodies or antibody fragments (e.g., antigen-binding fragments) comprise a VH domain having the amino acid sequence of SEQ ID NO : 13, 45, 77, or 608. In some embodiments, the invention also provides antibodies or antibody fragments (eg, antigen-binding fragments) that specifically bind to CCR7, wherein said antibodies or antibody fragments (eg, antigen-binding fragments) contain the CDR VH having the amino acid sequence of any of the VH CDRs listed in Tables 1 and 4 below. In specific embodiments, the present invention provides antibodies or antibody fragments (e.g., antigen-binding fragments) that specifically bind to CCR7, said antibodies containing (or alternatively consisting of) one, two, three, four, five, or more VH CDRs. having the amino acid sequence of any of the VH CDRs listed in Tables 1 and 4 below.

[00214] The present invention provides antibodies or antibody fragments (eg, antigen-binding fragments) that specifically bind to CCR7, wherein said antibodies or antibody fragments (eg, antigen-binding fragments) comprise a VL domain having the amino acid sequence of SEQ ID NO: 29 , 61, 93, or 624. The present invention also provides antibodies or antibody fragments (eg, antigen-binding fragments) that specifically bind to CCR7, wherein said antibodies or antibody fragments (eg, antigen-binding fragments) comprise a VL CDR having an amino acid sequence of any from the VL CDRs listed in Tables 1 and 4 below. In particular, the present invention provides antibodies or antibody fragments (e.g., antigen-binding fragments) that specifically bind to CCR7, wherein said antibodies or antibody fragments (e.g., antigen-binding fragments) comprise (or alternatively consist of) one, two, three or more VL CDRs having the amino acid sequence of any of the VL CDRs listed in Tables 1 and 4 below.

[00215] Other antibodies or antibody fragments (e.g., antigen-binding fragments) of the present invention include amino acids that have been mutated but have at least 60, 70, 80, 90, or 95 percent identity in the CDRs with the CDRs shown in the sequences described in Tables 1 and 4. In some embodiments, the antibodies comprise mutant amino acid sequences in which no more than 1, 2, 3, 4, or 5 amino acids have been mutated in the CDR regions compared to the CDR regions shown in the sequence described in tables 1 and 4.

[00216] The present invention also provides nucleic acid sequences that encode VH, VL, full length heavy chain and full length light chain of antibodies that specifically bind to CCR7. Such nucleic acid sequences can be optimized for expression in mammalian cells.

[00217] Throughout the text of this application, there may be a discrepancy between the description text and the sequence listing, with the description text taking precedence.

Table 1. Examples of anti-CCR7 antibodies of the present invention

506E15 (humanized CysMab-DAPA) SEQ ID NO: 1 HCDR1 (combined) GFTFSSYAMS SEQ ID NO: 2 HCDR2 (combined) TISSGGSFTYYPDSVKG SEQ ID NO: 3 HCDR3 (combined) RASTVVGTDFDV SEQ ID NO: 4 HDDR1 (Kabat) SYAMS SEQ ID NO: 5 HDDR2 (Kabat) TISSGGSFTYYPDSVKG SEQ ID NO: 6 HDDR3 (Kabat) RASTVVGTDFDV SEQ ID NO: 7 HCDR1 (Chothia) GFTFSSY SEQ ID NO: 8 HCDR2 (Chothia) SSGGSF SEQ ID NO: 9 HCDR3 (Chothia) RASTVVGTDFDV SEQ ID NO: 10 HCDR1 (IMGT) GFTFSSYA SEQ ID NO: 11 HDDR2 (IMGT) ISSGGSFT SEQ ID NO: 12 HDDR3 (IMGT) ARRASTVVGTDFDV SEQ ID NO: 13 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYAMSWIRQAPGKGLEWVATISSGGSFTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRASTVVGTDFDVWGQGTTVTVSS SEQ ID NO: 14 DNA VH GAAGTGCAGCTGGTGGAATCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACCTTCTCCAGCTACGCCATGTCCTGGATCCGGCAGGCCCCTGGCAAGGGACTGGAGTGGGTGGCCACCATCTCCTCCGGCGGCAGCTTCACCTACTACCCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGCCTCCACCGTCGTGGGCACCGATTTCGATGTGTGGGGCCAGGGCACAACCGTGACCGTGTCCTCC SEQ ID NO: 15 heavy chain
(DAPA, CysMab, mutations are underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYAMSWIRQAPGKGLEWVATISSGGSFTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRASTVVGTDFDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 16 heavy chain DNA GAAGTGCAGCTGGTGGAATCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACCTTCTCCAGCTACGCCATGTCCTGGATCCGGCAGGCCCCTGGCAAGGGACTGGAGTGGGTGGCCACCATCTCCTCCGGCGGCAGCTTCACCTACTACCCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGCCTCCACCGTCGTGGGCACCGATTTCGATGTGTGGGGCCAGGGCACAACCGTGACCGTGTCCTCCGCCTCCACCAAGGGACCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCCACCTCTGGCGGCACCGCCGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCCTGCCCTGTGACAGTGTCCTGGAACTCCGGCGCTCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACCGTGCCTTCCTCCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAAGTGGACAAGCGGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCCCCTGAGCTGCTGGGAGGCCCTTCCGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGATCCCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCCAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTGTCCAACAAGGCCCTGGCCG CTCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAAGTGTACACACTGCCTCCCAGCCGGGAAGAGATGACCAAGAACCAAGTGTCCCTGACCTGCCTCGTGAAGGGCTTCTACCCCTGCGATATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCTCCCGTGCTGGACAGCGACGGCTCATTCTTCCTGTACTCCAAGCTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGAGCCCCGGCAAG SEQ ID NO: 17 LCDR1 (combined) RASQDIGSSLN SEQ ID NO: 18 LCDR2 (combined) ATSSLDS SEQ ID NO: 19 LCDR3 (combined) LQYASSPPT SEQ ID NO: 20 LCDR1 (Kabat) RASQDIGSSLN SEQ ID NO: 21 LCDR2 (Kabat) ATSSLDS SEQ ID NO: 22 LCDR3 (Kabat) LQYASSPPT SEQ ID NO: 23 LCDR1 (Chothia) SQDIGSS SEQ ID NO: 24 LCDR2 (Chothia) ATS SEQ ID NO: 25 LCDR3 (Chothia) YASSPP SEQ ID NO: 26 LCDR1 (IMGT) QDIGSS SEQ ID NO: 27 LCDR2 (IMGT) ATS SEQ ID NO: 28 LCDR3 (IMGT) LQYASSPPT SEQ ID NO: 29 VL DIQMTQSPSSLSASVGDRVTLTCRASQDIGSSLNWLQQKPGKAIKRLIYATSSLDSGVPSRFSGSRSGTDYTLTISSLQPEDFVVYYCLQYASSPPTFGGGTKLEIK SEQ ID NO: 30 DNA VL GACATCCAGATGACCCAGAGCCCCTCCAGCCTGTCCGCCTCCGTGGGCGATAGAGTGACCCTGACCTGCCGGGCCTCCCAGGACATCGGCTCCTCCCTGAACTGGCTGCAGCAGAAGCCCGGCAAGGCCATCAAGCGGCTGATCTACGCCACCTCCTCCCTGGACTCCGGCGTGCCCTCCCGGTTCTCTGGCTCCAGATCCGGCACCGACTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGTGGTGTACTACTGCCTGCAGTACGCCTCCAGCCCTCCCACCTTCGGCGGAGGCACCAAGCTGGAAATCAAG SEQ ID NO: 31 light chain DIQMTQSPSSLSASVGDRVTLTCRASQDIGSSLNWLQQKPGKAIKRLIYATSSLDSGVPSRFSGSRSGTDYTLTISSLQPEDFVVYYCLQYASSPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 32 light chain DNA GACATCCAGATGACCCAGAGCCCCTCCAGCCTGTCCGCCTCCGTGGGCGATAGAGTGACCCTGACCTGCCGGGCCTCCCAGGACATCGGCTCCTCCCTGAACTGGCTGCAGCAGAAGCCCGGCAAGGCCATCAAGCGGCTGATCTACGCCACCTCCTCCCTGGACTCCGGCGTGCCCTCCCGGTTCTCTGGCTCCAGATCCGGCACCGACTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGTGGTGTACTACTGCCTGCAGTACGCCTCCAGCCCTCCCACCTTCGGCGGAGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC 121G12 (humanized CysMab, DAPA) SEQ ID NO: 33 HCDR1 (combined) GFTFSTYAMS SEQ ID NO: 34 HCDR2 (combined) TISDAGSYSYYPDNVKG SEQ ID NO: 35 HCDR3 (combined) RGSRYEEYYVMDY SEQ ID NO: 36 HDDR1 (Kabat) TYAMS SEQ ID NO: 37 HDDR2 (Kabat) TISDAGSYSYYPDNVKG SEQ ID NO: 38 HDDR3 (Kabat) RGSRYEEYYVMDY SEQ ID NO: 39 HCDR1 (Chothia) GFTFSTY SEQ ID NO: 40 HCDR2 (Chothia) SDAGSY SEQ ID NO: 41 HCDR3 (Chothia) RGSRYEEYYVMDY SEQ ID NO: 42 HCDR1 (IMGT) GFTFSTYA SEQ ID NO: 43 HDDR2 (IMGT) ISDAGSYS SEQ ID NO: 44 HDDR3 (IMGT) ARRGSRYEEYYVMDY SEQ ID NO: 45 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSTYAMSWVRQAPGKGLEWVATISDAGSYSYYPDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGSRYEEYYVMDYWGQGTTVTVSS SEQ ID NO: 46 DNA VH GAAGTGCAGCTGGTGGAATCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACCTTCTCCACCTACGCCATGTCCTGGGTCCGACAGGCCCCTGGAAAGGGCCTGGAGTGGGTGGCCACCATCTCCGACGCCGGCTCCTACTCCTACTACCCCGACAACGTGAAGGGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAGACGGGGCTCCAGATACGAAGAGTACTACGTGATGGACTACTGGGGCCAGGGCACAACCGTGACCGTGTCCTCC SEQ ID NO: 47 heavy chain
(DAPA, CysMab, mutations are underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSTYAMSWVRQAPGKGLEWVATISDAGSYSYYPDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGSRYEEYYVMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 48 heavy chain DNA GAAGTGCAGCTGGTGGAATCTGGCGGCGGACTGGTCAAGCCTGGCGGCTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACCTTCTCCACCTACGCCATGTCCTGGGTCCGACAGGCCCCTGGAAAGGGCCTGGAGTGGGTGGCCACCATCTCCGACGCCGGCTCCTACTCCTACTACCCCGACAACGTGAAGGGCCGGTTCACCATCTCCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAGACGGGGCTCCAGATACGAAGAGTACTACGTGATGGACTACTGGGGCCAGGGCACAACCGTGACCGTGTCCTCCGCCTCCACCAAGGGACCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCCACCTCTGGCGGCACCGCCGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCCTGCCCTGTGACAGTGTCCTGGAACTCCGGCGCTCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACCGTGCCTTCCTCCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAAGTGGACAAGCGGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCCCCTGAGCTGCTGGGAGGCCCTTCCGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGATCCCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCCAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTGTCCAACAAGGCCCTGG CCGCTCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAAGTGTACACACTGCCTCCCAGCCGGGAAGAGATGACCAAGAACCAAGTGTCCCTGACCTGCCTCGTGAAGGGCTTCTACCCCTGCGATATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCTCCCGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCCAAGCTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGAGCCCCGGCAAG SEQ ID NO: 49 LCDR1 (combined) RASQSISNNLH SEQ ID NO: 50 LCDR2 (combined) YASQSIS SEQ ID NO: 51 LCDR3 (combined) QQSSSWLT SEQ ID NO: 52 LCDR1 (Kabat) RASQSISNNLH SEQ ID NO: 53 LCDR2 (Kabat) YASQSIS SEQ ID NO: 54 LCDR3 (Kabat) QQSSSWLT SEQ ID NO: 55 LCDR1 (Chothia) SQSISNN SEQ ID NO: 56 LCDR2 (Chothia) YAS SEQ ID NO: 57 LCDR3 (Chothia) SSSWL SEQ ID NO: 58 LCDR1 (IMGT) QSISNN SEQ ID NO: 59 LCDR2 (IMGT) YAS SEQ ID NO: 60 LCDR3 (IMGT) QQSSSWLT SEQ ID NO: 61 VL EIVLTQSPATLSVSPGERVTLSCRASQSISNNLHWYQQKPGQAPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSVEPEDFGVYFCQQSSSWLTFGQGTKLEIK SEQ ID NO: 62 DNA VL GAGATCGTGCTGACACAGTCCCCTGCCACCCTGTCTGTGTCTCCCGGCGAGAGAGTGACCCTGTCCTGCCGGGCCTCCCAGTCCATCTCCAACAACCTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATTAAGTACGCCTCCCAGAGCATCTCCGGCATCCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACCCTGACCATCTCCAGCGTGGAACCCGAGGACTTCGGCGTGTACTTCTGCCAGCAGTCCTCCTCCTGGCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG SEQ ID NO: 63 light chain EIVLTQSPATLSVSPGERVTLSCRASQSISNNLHWYQQKPGQAPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSVEPEDFGVYFCQQSSSWLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACENSFTHQGL SEQ ID NO: 64 light chain DNA GAGATCGTGCTGACACAGTCCCCTGCCACCCTGTCTGTGTCTCCCGGCGAGAGAGTGACCCTGTCCTGCCGGGCCTCCCAGTCCATCTCCAACAACCTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATTAAGTACGCCTCCCAGAGCATCTCCGGCATCCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACCCTGACCATCTCCAGCGTGGAACCCGAGGACTTCGGCGTGTACTTCTGCCAGCAGTCCTCCTCCTGGCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC 674J13 (humanized, CysMab, DAPA) SEQ ID NO: 65 HCDR1 (combined) GYSITSGYSWH SEQ ID NO: 66 HCDR2 (combined) HIHSSGSTNYNPSLKS SEQ ID NO: 67 HCDR3 (combined) GGVQAFAY SEQ ID NO: 68 HDDR1 (Kabat) SGYSWH SEQ ID NO: 69 HDDR2 (Kabat) HIHSSGSTNYNPSLKS SEQ ID NO: 70 HDDR3 (Kabat) GGVQAFAY SEQ ID NO: 71 HCDR1 (Chothia) GYSITSGY SEQ ID NO: 72 HCDR2 (Chothia) HSSGS SEQ ID NO: 73 HCDR3 (Chothia) GGVQAFAY SEQ ID NO: 74 HCDR1 (IMGT) GYSITSGYS SEQ ID NO: 75 HDDR2 (IMGT) IHSSGST SEQ ID NO: 76 HDDR3 (IMGT) ARGGVQAFAY SEQ ID NO: 77 vh DVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYSWHWIRQHPGKGLEWMAHIHSSGSTNYNPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARGGVQAFAYWGQGTLVTVSS SEQ ID NO: 78 DNA VH GACGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTCAAGCCCTCCCAGACCCTGTCCCTGACCTGCACCGTGTCCGGCTACTCTATCACCTCCGGCTACAGCTGGCACTGGATCCGGCAGCACCCCGGCAAGGGCCTGGAATGGATGGCCCACATCCACTCCTCCGGCTCCACCAACTACAACCCCAGCCTGAAGTCCCGGATCACCATCTCCCGGGACACCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCCGCTGACACCGCCGTGTACTACTGTGCCAGAGGCGGCGTGCAGGCCTTCGCTTATTGGGGCCAGGGAACCCTGGTCACCGTGTCCTCC SEQ ID NO: 79 heavy chain
(DAPA, CysMab, mutations are underlined) DVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYSWHWIRQHPGKGLEWMAHIHSSGSTNYNPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARGGVQAFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 80 heavy chain DNA GACGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTCAAGCCCTCCCAGACCCTGTCCCTGACCTGCACCGTGTCCGGCTACTCTATCACCTCCGGCTACAGCTGGCACTGGATCCGGCAGCACCCCGGCAAGGGCCTGGAATGGATGGCCCACATCCACTCCTCCGGCTCCACCAACTACAACCCCAGCCTGAAGTCCCGGATCACCATCTCCCGGGACACCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCCGCTGACACCGCCGTGTACTACTGTGCCAGAGGCGGCGTGCAGGCCTTCGCTTATTGGGGCCAGGGAACCCTGGTCACCGTGTCCTCCGCCAGCACCAAGGGACCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCCACCTCTGGCGGCACCGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCTGCCCCGTGACCGTGTCCTGGAACTCCGGCGCTCTGACCTCCGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCAGCGTCGTGACCGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAAGTGGACAAGCGGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCCCCTGAGCTGCTGGGAGGCCCTTCCGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGATCCCGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCCAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTGTCCAACAAGGCCCTGGCCGCTCCCATCGAAA AGACCATCTCCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAAGTGTACACACTGCCTCCCAGCCGGGAAGAGATGACCAAGAATCAAGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCCTGCGATATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCTCCCGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCCAAGCTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGTCCCCTGGCAAG SEQ ID NO: 81 LCDR1 (combined) SASSSVIYMH SEQ ID NO: 82 LCDR2 (combined) DTSKLAS SEQ ID NO: 83 LCDR3 (combined) QQWSSNPLT SEQ ID NO: 84 LCDR1 (Kabat) SASSSVIYMH SEQ ID NO: 85 LCDR2 (Kabat) DTSKLAS SEQ ID NO: 86 LCDR3 (Kabat) QQWSSNPLT SEQ ID NO: 87 LCDR1 (Chothia) SSSVIY SEQ ID NO: 88 LCDR2 (Chothia) DTS SEQ ID NO: 89 LCDR3 (Chothia) WSSNPL SEQ ID NO: 90 LCDR1 (IMGT) SSVIY SEQ ID NO: 91 LCDR2 (IMGT) DTS SEQ ID NO: 92 LCDR3 (IMGT) QQWSSNPLT SEQ ID NO: 93 VL EIVLTQSPATLSASPGERVTMSCSASSSVIYMHWYQQKPGQAPRRWIYDTSKLASGVPARFSGSGSGTDYTLTISSMEPEDAAVYYCQQWSSNPLTFGQGTKLEIK SEQ ID NO: 94 DNA VL GAGATCGTGCTGACACAGTCCCCTGCCACCCTGTCCGCCTCTCCAGGCGAGCGCGTGACAATGTCCTGCTCCGCCTCCTCCTCCGTGATCTACATGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCGGTGGATCTACGATACCTCCAAGCTGGCCTCCGGCGTGCCCGCCAGATTCTCCGGCTCTGGCTCTGGCACCGACTACACCCTGACCATCTCCAGCATGGAACCCGAGGACGCCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCTCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG SEQ ID NO: 95 light chain EIVLTQSPATLSASPGERVTMSCSASSSVIYMHWYQQKPGQAPRRWIYDTSKLASGVPARFSGSGSGTDYTLTISSMEPEDAAVYYCQQWSSNPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS SEQ ID NO: 96 light chain DNA GAGATCGTGCTGACACAGTCCCCTGCCACCCTGTCCGCCTCTCCAGGCGAGCGCGTGACAATGTCCTGCTCCGCCTCCTCCTCCGTGATCTACATGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCGGTGGATCTACGATACCTCCAAGCTGGCCTCCGGCGTGCCCGCCAGATTCTCCGGCTCTGGCTCTGGCACCGACTACACCCTGACCATCTCCAGCATGGAACCCGAGGACGCCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCTCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC

[00218] Other antibodies of the present invention include antibodies in which the amino acids or nucleic acids encoding amino acids have been mutated, but they are characterized by at least 60, 70, 80, 90, or 95 percent identity with the sequences described in tables 1 and 4. In some embodiments, 1, 2, 3, 4, or 5 amino acids have been mutated in the variable regions compared to the variable regions shown in the sequence described in Tables 1 and 4, while retaining substantially the same therapeutic activity. as the antibodies listed in Tables 1 and 4.

[00219] Since each of these antibodies can bind to CCR7 sequences (amino acid sequences and nucleotide sequences encoding amino acid sequences), VH, VL, full length light chain and full length heavy chain can be "mix and match" to create other CCR7-binding antibodies along the present invention. Such "mix and match" CCR7 binding antibodies can be tested using binding assays known in the art (eg, ELISAs and other assays described in the Examples section). When such chains are subjected to mixing and matching, the VH sequence from a particular VH/VL pair should be replaced with a structurally similar VH sequence. Similarly, a full length heavy chain sequence from a particular full length heavy chain/full length light chain pair should be replaced with a structurally similar full length heavy chain sequence. Similarly, a VL sequence from a particular VH/VL pair should be replaced with a structurally similar VL sequence. Similarly, a full length light chain sequence from a particular full length heavy chain/full length light chain pair should be replaced with a structurally similar full length light chain sequence. Accordingly, in one aspect of the present invention, there is provided an isolated monoclonal antibody, or antigen-binding region thereof, having: a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 13, 45, 77, and 608; and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 29, 61, 93 and 624; where the antibody specifically binds to CCR7.

[00220] In another aspect of the present invention, there are: (i) an isolated monoclonal antibody having: a full-length heavy chain containing an amino acid sequence that has been optimized for expression in an expression system that is a mammalian cell selected from the group consisting of SEQ ID NO : 15, 47, 79 and 610; and a full length light chain containing an amino acid sequence that has been optimized for expression in a mammalian cell selected from the group consisting of SEQ ID NOs: 31, 63, 95 and 626; or (ii) a functional protein containing an antigen-binding portion thereof.

[00221] In another aspect of the present invention, CCR7 binding antibodies are provided that comprise the heavy chain and light chain CDR1, CDR2, and CDR3 described in Tables 1 and 4, or combinations thereof. The amino acid sequences of CDR1 VH antibodies are shown, for example, under SEQ ID NOs: 1, 4, 7, 10, 33, 36, 39, 42, 65, 68, 71, and 74. The amino acid sequences of CDR2 VH antibodies are shown, for example, under SEQ ID NOs: 2, 5, 8, 11, 34, 37, 40, 43, 66, 69, 72, and 75. The amino acid sequences of CDR3 VH antibodies are shown, for example, under SEQ ID NOs: 3, 6, 9, 12, 35, 38, 41, 44, 67, 70, 73 and 76. The amino acid sequences of CDR1 VL antibodies are shown, for example, under SEQ ID NOs: 17, 20, 23, 26, 49, 52, 55, 58, 81, 84, 87 and 90. The amino acid sequences of CDR2 VL antibodies are shown, for example, under SEQ ID NOs: 18, 21, 24, 27, 50, 53, 56, 59, 82, 85, 88, and 91. The amino acid sequences of CDR3 VL antibodies are shown, for example, under SEQ ID NOs: 19, 22, 25, 28, 51, 54, 57, 60, 83, 86, 89 and 92.

[00222] Given that each of these antibodies can bind to CCR7, and that antigen binding specificity is primarily provided by the CDR1, 2 and 3 regions, the CDR1, CDR2 and CDR3 VH sequences and the CDR1, CDR2 and CDR3 VL sequences can be "mix and match" (i.e. CDRs from different antibodies can be mixed and matched. Such "mix and match" CCR7 binding antibodies can be tested using the binding assays known in the art and described in the Examples (e.g. , ELISA species).When VH CDR sequences are subjected to mixing and matching, the sequence of CDR1, CDR2 and/or CDR3 from a particular VH sequence should be replaced with structurally similar CDR sequence(s). Similarly, when VL CDR sequences are subjected to mixing and selection, the CDR1, CDR2 and/or CDR3 sequence from a particular VL sequence should be replaced with a structurally similar(- th) CDR sequence(s). One of ordinary skill in the art will appreciate that new VH and VL sequences can be created by replacing one or more VH and/or VL CDR region sequences with structurally similar sequences from the CDR sequences shown herein for the monoclonal antibodies of the present invention.

[00223] Accordingly, in some embodiments, the invention provides an isolated monoclonal antibody or antigen-binding region thereof comprising a heavy chain CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 4, 7, 10, 33, 36 , 39, 42, 65, 68, 71, 74, 596, 599, 602 and 605; a heavy chain CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 5, 8, 11, 34, 37, 40, 43, 66, 69, 72, 75, 597, 600, 603, and 606; a heavy chain CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 3, 6, 9, 12, 35, 38, 41, 44, 67, 70, 73, 76, 598, 601, 604 and 607; A light chain CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 17, 20, 23, 26, 49, 52, 55, 58, 81, 84, 87, 90, 612, 615, 618 and 621; A light chain CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 18, 21, 24, 27, 50, 53, 56, 59, 82, 85, 88, 91, 613, 616, 619, and 622; and a light chain CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 19, 22, 25, 28, 51, 54, 57, 60, 83, 86, 89, 92, 614, 617, 620 and 623 ; where the antibody specifically binds CCR7.

[00224] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:1, a heavy chain CDR2 of SEQ ID NO:2; heavy chain CDR3 under SEQ ID NO:3; Light chain CDR1 under SEQ ID NO:17; Light chain CDR2 of SEQ ID NO:18 and light chain CDR3 of SEQ ID NO:19.

[00225] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:4, a heavy chain CDR2 of SEQ ID NO:5; heavy chain CDR3 under SEQ ID NO:6; Light chain CDR1 under SEQ ID NO:20; Light chain CDR2 of SEQ ID NO:21 and Light chain CDR3 of SEQ ID NO:22.

[00226] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:7, a heavy chain CDR2 of SEQ ID NO:8; heavy chain CDR3 under SEQ ID NO:9; Light chain CDR1 under SEQ ID NO:23; Light chain CDR2 of SEQ ID NO:24 and Light chain CDR3 of SEQ ID NO:25.

[00227] In a specific embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:10, a heavy chain CDR2 of SEQ ID NO:11; heavy chain CDR3 under SEQ ID NO:12; Light chain CDR1 under SEQ ID NO:26; Light chain CDR2 of SEQ ID NO:27 and Light chain CDR3 of SEQ ID NO:28.

[00228] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:33, a heavy chain CDR2 of SEQ ID NO:34; heavy chain CDR3 under SEQ ID NO:35; Light chain CDR1 under SEQ ID NO:49; Light chain CDR2 of SEQ ID NO:50 and Light chain CDR3 of SEQ ID NO:51.

[00229] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:36, a heavy chain CDR2 of SEQ ID NO:37; heavy chain CDR3 under SEQ ID NO:38; Light chain CDR1 under SEQ ID NO:52; Light chain CDR2 of SEQ ID NO:53 and Light chain CDR3 of SEQ ID NO:54.

[00230] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:39, a heavy chain CDR2 of SEQ ID NO:40; heavy chain CDR3 under SEQ ID NO:41; Light chain CDR1 under SEQ ID NO:55; The light chain CDR2 of SEQ ID NO:56 and the light chain CDR3 of SEQ ID NO:57.

[00231] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:42, a heavy chain CDR2 of SEQ ID NO:43; heavy chain CDR3 under SEQ ID NO:44; Light chain CDR1 under SEQ ID NO:58; Light chain CDR2 of SEQ ID NO:59 and light chain CDR3 of SEQ ID NO:60.

[00232] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:65, a heavy chain CDR2 of SEQ ID NO:66; heavy chain CDR3 of SEQ ID NO:67; Light chain CDR1 under SEQ ID NO:81; Light chain CDR2 of SEQ ID NO:82 and Light chain CDR3 of SEQ ID NO:83.

[00233] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:68, a heavy chain CDR2 of SEQ ID NO:69; heavy chain CDR3 under SEQ ID NO:70; Light chain CDR1 under SEQ ID NO:84; Light chain CDR2 of SEQ ID NO:85 and Light chain CDR3 of SEQ ID NO:86.

[00234] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:71, a heavy chain CDR2 of SEQ ID NO:72; heavy chain CDR3 under SEQ ID NO:73; Light chain CDR1 under SEQ ID NO:87; Light chain CDR2 of SEQ ID NO:88 and Light chain CDR3 of SEQ ID NO:89.

[00235] In a particular embodiment, the antibody or antibody fragment (eg, antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain CDR1 of SEQ ID NO:74, a heavy chain CDR2 of SEQ ID NO:75; heavy chain CDR3 under SEQ ID NO:76; Light chain CDR1 under SEQ ID NO:90; Light chain CDR2 of SEQ ID NO:91 and Light chain CDR3 of SEQ ID NO:92.

[00236] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region that contains HCDR1 of SEQ ID NO:596, HCDR2 of SEQ ID NO:597, and HCDR3 of SEQ ID NO:597. SEQ ID NO:598; and a light chain variable region that contains LCDR1 of SEQ ID NO:612, LCDR2 of SEQ ID NO:613, and LCDR3 of SEQ ID NO:614.

[00237] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region that contains HCDR1 of SEQ ID NO:599, HCDR2 of SEQ ID NO:600, and HCDR3 of SEQ ID NO:600. SEQ ID NO:601; and a light chain variable region that contains LCDR1 of SEQ ID NO:615, LCDR2 of SEQ ID NO:616, and LCDR3 of SEQ ID NO:617.

[00238] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region that contains HCDR1 of SEQ ID NO:602, HCDR2 of SEQ ID NO:603, and HCDR3 of SEQ ID NO:603. SEQ ID NO:604; and a light chain variable region that contains LCDR1 of SEQ ID NO:618, LCDR2 of SEQ ID NO:619, and LCDR3 of SEQ ID NO:620.

[00239] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region that contains HCDR1 of SEQ ID NO:605, HCDR2 of SEQ ID NO:606, and HCDR3 of SEQ ID NO:606. SEQ ID NO:607; and a light chain variable region that contains LCDR1 of SEQ ID NO:621, LCDR2 of SEQ ID NO:622, and LCDR3 of SEQ ID NO:623.

[00240] In a specific embodiment, an antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:13 and a light chain variable region (VL ) containing the amino acid sequence under SEQ ID NO:29.

[00241] In a particular embodiment, an antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:45 and a light chain variable region (VL ) containing the amino acid sequence under SEQ ID NO:61.

[00242] In a particular embodiment, an antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:77 and a light chain variable region (VL ) containing the amino acid sequence under SEQ ID NO:93.

[00243] In a particular embodiment, an antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:608 and a light chain variable region (VL ) containing the amino acid sequence under SEQ ID NO:624.

[00244] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain containing the amino acid sequence of SEQ ID NO:15 and a light chain containing the amino acid sequence of SEQ ID NO: 31.

[00245] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain containing the amino acid sequence of SEQ ID NO:47 and a light chain containing the amino acid sequence of SEQ ID NO: 63.

[00246] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain containing the amino acid sequence of SEQ ID NO:79 and a light chain containing the amino acid sequence of SEQ ID NO: 95.

[00247] In a specific embodiment, the antibody or antibody fragment (e.g., antigen-binding fragments) that specifically binds to CCR7 comprises a heavy chain containing the amino acid sequence of SEQ ID NO:610 and a light chain containing the amino acid sequence of SEQ ID NO: 626

[00248] In some embodiments, an antibody that specifically binds to CCR7 is an antibody or antibody fragment (e.g., an antigen-binding fragment) as described in Tables 1 and 4.

1. Identification of epitopes and antibodies that bind to the same epitope

[00249] The present invention also provides antibodies and antibody fragments (e.g., antigen-binding fragments) that specifically bind to the same epitope as the anti-CCR7 antibodies described in Tables 1 and 4 or cross-compete with the antibodies described in Tables 1 and 4. Tables 1 and 4. Therefore, additional antibodies and antibody fragments (e.g., antigen-binding fragments) can be identified based on their ability to cross-compete (e.g., competitive binding inhibition, in a statistically significant manner) with other antibodies of the present invention in CCR7 binding assays, for example, using BIACORE or assays known to those skilled in the art to measure binding. The ability of a test antibody to inhibit the binding of antibodies and antibody fragments (e.g., antigen-binding fragments) of the present invention to CCR7 (e.g., human CCR7) demonstrates that the test antibody can compete with such antibody or antibody fragment (e.g., antigen-binding fragments) for binding to CCR7; however, according to a non-limiting theory, such an antibody can bind to the same or related (for example, structurally similar or spatially close or overlapping) epitope on the CCR7 protein as the antibody or antibody fragment (for example, antigen-binding fragments) with which it competes. In some embodiments, antibodies that bind to the same epitope on CCR7 as the antibodies or antibody fragments (eg, antigen-binding fragments) described in Tables 1 and 4 are human or humanized monoclonal antibodies. Such human or humanized monoclonal antibodies may be prepared and isolated as described herein.

2. Additional modification of the framework region in the Fc region

[00250] The immunoconjugates of the present invention may contain modified antibodies or antigen-binding fragments thereof, which further contain modifications to framework residues within the VH and/or VL, for example, to improve the properties of the antibody. In some embodiments, modifications to the framework region are made to reduce the immunogenicity of the antibody. For example, one approach is for one or more framework residues to "mutate back" in the direction of the corresponding germline sequence. More specifically, an antibody that has undergone a somatic mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the framework sequences of the antibody with the germline sequences from which the antibody is derived. To return the framework region sequences to their germline configuration, somatic mutations can be introduced to "mutate back" into the germline sequence, for example, by site-directed mutagenesis. It is implied that such subjected to "mutation to the original form" antibodies are also covered by the present invention.

[00251] Another type of framework modification involves the mutation of one or more residues within the framework, or even within one or more CDR regions, to remove T-cell epitopes, thereby reducing the potential immunogenicity of the antibody. This approach is also referred to as "deimmunization" and is described in more detail in US Patent Application Publication No. 20030153043 by Carr et al .

[00252] As an addition or alternative to modifications made within the framework or CDR regions, the antibodies of the present invention can be designed to include modifications within the Fc region, typically to alter one or more of the functional properties of the antibody, such as as serum half-life, complement fixation, Fc receptor binding and/or antigen-dependent cellular cytotoxicity (ADCC). In addition, the antibody of the present invention may be chemically modified (for example, one or more chemical moieties may be attached to the antibody), or it may be modified to change its glycosylation pattern, again to alter one or more of the functional properties of the antibody. Each of these embodiments is described in more detail below.

[00253] In one embodiment, the CH1 hinge is modified such that the number of cysteine residues in the hinge changes, eg, increases or decreases. This approach is further described in US Pat. No. 5,677,425 to Bodmer et al . The number of cysteine residues in the CH1 hinge region is changed, for example, to facilitate assembly of the light and heavy chains, or to increase or decrease the stability of the antibody.

[00254] In some embodiments, an antibody or antibody fragment disclosed herein contains modified or engineered amino acid residues, e.g., one or more cysteine residues, as sites for conjugation to a drug moiety (Junutula JR, et al., Nat Biotechnol 2008, 26:925-932). In one embodiment, the present invention provides a modified antibody or antibody fragment containing the replacement of one or more amino acids with cysteine at the positions described herein. Cysteine substitution sites are located in the constant regions of an antibody or antibody fragment and are thus applicable to a variety of antibodies or antibody fragments, and the sites are chosen to provide stable and uniform conjugates. The modified antibody or fragment may have one, two or more cysteine substitutions, and such substitutions may be used in combination with other modification and conjugation methods described herein. Methods for inserting cysteine at specific antibody locations are known in the art, see for example Lyons et al , (1990) Protein Eng., 3:703-708, WO 2011/005481, WO2014/124316, WO 2015/138615. In some embodiments, the modified antibody comprises a substitution of one or more amino acids for cysteine in its constant region at a position selected from positions 117, 119, 121, 124, 139, 152, 153, 155, 157, 164, 169, 171, 174, 189, 191, 195, 197, 205, 207, 246, 258, 269, 274, 286, 288, 290, 292, 293, 320, 322, 326, 333, 334, 335, 337, 344, 355, 360 375, 382, 390, 392, 398, 400 and 422 of the heavy chain of the antibody, and where the position numbering follows the EU system. In some embodiments, the modified antibody or antibody fragment comprises a replacement of one or more amino acids with a cysteine in its constant region at a position selected from positions 107, 108, 109, 114, 129, 142, 143, 145, 152, 154, 156, 159 , 161, 165, 168, 169, 170, 182, 183, 197, 199, and 203 of the light chain of an antibody or antibody fragment, where the position numbering follows the EU system, and where the light chain is a human kappa light chain. In some embodiments, the modified antibody or antibody fragment comprises a combination of two or more amino acid substitutions for cysteine in its constant regions, where the combinations include substitutions at positions 375 of the antibody heavy chain, position 152 of the antibody heavy chain, position 360 of the antibody heavy chain, or position 107 the light chain of the antibody, and where the position numbering follows the EU system. In some embodiments, the modified antibody or antibody fragment comprises a single amino acid substitution for cysteine in its constant regions, where the substitution is at position 375 of the antibody heavy chain, position 152 of the antibody heavy chain, position 360 of the antibody heavy chain, position 107 of the antibody light chain, position 165 of the antibody light chain or position 159 of the antibody light chain, and where the position numbering is according to the EU system, and where the light chain is a kappa chain. In specific embodiments, the modified antibody or antibody fragment contains a combination of two amino acid substitutions for cysteine in its constant regions, where the combinations include substitutions at positions 375 of the antibody heavy chain and position 152 of the antibody heavy chain, where the position numbering follows the EU system. In specific embodiments, the modified antibody or fragment of such an antibody comprises a single amino acid substitution for a cysteine at position 360 of the heavy chain of the antibody, where position numbering follows the EU system. In other specific embodiments, the modified antibody or antibody fragment comprises a single amino acid substitution for cysteine at position 107 of the antibody light chain, and where the position numbering follows the EU system, and where the light chain is a kappa chain.

[00255] In additional embodiments, the antibodies or antibody fragments (e.g., antigen binding fragment) useful in the immunoconjugates of the present invention include modified or engineered antibodies, such as an antibody modified to introduce one or more other reactive amino acids (other than cysteine), including Pcl, pyrrolysin, peptide tags (such as S6, A1, and ybbR tags), and non-natural amino acids, in place of at least one amino acid from the native sequence, thereby providing a reactive site on the antibody or antigen-binding fragment for conjugation to the fragment, which is drug, or a linker-drug moiety with a complementary reactive group. For example, antibodies or antibody fragments can be modified to introduce Pcl or pyrrolysine (W. Ou, et al., (2011) PNAS 108 (26), 10437-10442; WO2014124258) or unnatural amino acids (JY Axup, et al., Proc Natl Acad Sci USA, 109 (2012), pp. 16101-16106, review see CC Liu and PG Schultz (2010) Annu Rev Biochem 79, 413-444, CH Kim, et al., (2013) Curr Opin Chem Biol. 17, 412-419) as drug conjugation sites. Similarly, peptide labels for enzymatic conjugation methods can be introduced into an antibody (Strop P., et al., Chem Biol. 2013, 20(2):161-7; Rabuka D., Curr Opin Chem Biol. 2010 Dec;14( 6):790-6; Rabuka D, et al., Nat Protoc. 2012, 7(6):1052-67). One other example is the use of 4'-phosphopantetheinyltransferases (PPTase) for the conjugation of coenzyme A analogs (WO2013184514) and (Grünewald et al., (2015) Bioconjugate Chem. 26 (12), 2554-62). Methods for conjugating such modified or engineered antibodies to payloads or linker-payload combinations are known in the art.

[00256] In another embodiment, to reduce the biological half-life of an antibody, the Fc-hinge region of the antibody is mutated. More specifically, one or more amino acid mutations are introduced in the border region between the CH2-CH3 domains of the Fc-hinge fragment, whereby the antibody has impaired binding to Staphylococcus A protein (SpA) compared to the binding of the native Fc-hinge domain to SpA. This approach is described in more detail in US patent No. 6165745 from Ward et al .

[00257] In other embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector functions of the antibody. For example, one or more amino acids can be replaced with a different amino acid residue such that the antibody has an altered affinity for the effector ligand but retains the ability to bind the parent antibody's antigen. The effector ligand for which the affinity is changed may be, for example, the Fc receptor or the C1 component of complement. This approach is described, for example, in US patent No. 5624821 and 5648260, both from Winter et al .

[00258] In another embodiment, one or more amino acids selected from amino acid residues can be replaced with a different amino acid residue, whereby the antibody has altered C1q binding and/or reduced or eliminated complement-dependent cytotoxicity (CDC). This approach is described, for example, in US patent No. 6194551 from Idusogie et al .

[00259] In another embodiment, one or more amino acid residues are altered to thereby alter the ability of the antibody to fix complement. This approach is described, for example, in PCT publication WO 94/29351 from Bodmer et al. Allotypic amino acid residues include, without limitation, the heavy chain constant region of the IgG1, IgG2, and IgG3 subclasses, as well as the light chain constant region of the kappa chain isotypes, as described in Jefferis et al., MAbs. 1:332-338 (2009).

[00260] Fusion protein-based complexes with an antibody containing such mutations mediate reduced or absent antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). In some embodiments, amino acid residues L234 and L235 in the IgG1 constant region are changed to A234 and A235. In some embodiments, the N267 amino acid residue in the IgG1 constant region is replaced by A267. In some embodiments, amino acid residues D265 and P329 in the IgG1 constant region are changed to A265 and A329. In some embodiments, the immunoglobulin heavy chain optionally contains a mutation or combination of mutations conferring reduced effector function, selected from any of D265A, P329A, P329G, N297A, D265A/P329A, D265A/N297A, L234/L235A, P329A/L234A/L235A, and P329G /L234A/L235A. In specific embodiments, the immunoconjugate comprises an immunoglobulin heavy chain containing a mutation or combination of mutations conferring reduced effector function selected from any of D265A, P329A, P329G, N297A, D265A/P329A, D265A/N297A, L234/L235A, P329A/L234A/L235A and P329G/L234A/L235A.

[00261] In another embodiment, one or more amino acid residues are altered to thereby alter the ability of the antibody to fix complement. This approach is described, for example, in PCT publication WO 94/29351 from Bodmer et al. In a specific embodiment, one or more amino acids of an antibody or antigen-binding fragment of the present invention are replaced with one or more allotypic amino acid residues. Allotypic amino acid residues also include, without limitation, the heavy chain constant region of the IgG1, IgG2, and IgG3 subclasses, as well as the light chain constant region of the kappa chain isotype, as described in Jefferis et al., MAbs. 1:332-338 (2009).

[00262] In yet another embodiment, the glycosylation pattern of the antibody is modified. For example, an aglycosylated antibody (ie, an antibody that lacks glycosylation) can be produced. The pattern of glycosylation can be altered, for example, to increase the affinity of the antibody for the "antigen". Such carbohydrate modifications can be made, for example, by changing one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made that result in the elimination of one or more glycosylation sites in the framework region of the variable region, thereby eliminating glycosylations at that site. Such aglycosylation can increase the affinity of the antibody for the antigen. This approach is described, for example, US patent No. 5714350 and 6350861 from Co et al .

[00263] In another embodiment, the antibody is modified to increase its biological half-life. Various approaches are possible. For example, you can enter one or more of the following mutations: T252L, T254S, T256F, as described in US patent No. 6277375 from Ward. Alternatively, to increase the biological half-life, the antibody can be modified within the CH1 or CL region such that it contains an epitope for binding to the reutilization receptor taken from two loops of the CH2 domain from the IgG Fc region, as described in US Pat. Nos. 5,869,046 and 6121022 from Presta et al .

3. Production of antibodies to CCR7

[00264] Anti-CCR7 antibodies and fragments of such antibodies (for example, antigen-binding fragments) can be obtained using any means known in the art, including, without limitation, recombinant expression, chemical synthesis, and enzymatic cleavage of antibody tetramers, while full-length monoclonal antibodies can be obtained, for example, using hybrid or recombinant production. Recombinant expression can be carried out in any suitable host cells known in the art, for example, mammalian host cells, bacterial host cells, yeast host cells, insect host cells, etc. d.

[00265] The present invention further provides polynucleotides encoding the antibodies described herein, for example, polynucleotides encoding variable regions or heavy or light chain segments containing the complementarity determining regions described herein. In some embodiments, a polynucleotide encoding heavy chain variable regions is characterized by at least 85%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or 100% nucleic acid sequence identity with a polynucleotide selected from the group consisting of SEQ ID NOs: 14, 46, 78, and 609. In some embodiments, a polynucleotide encoding light chain variable regions is characterized by at least 85%, 89% , 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% nucleic acid sequence identity with a polynucleotide selected from the group consisting of SEQ ID NO: 30, 62, 94 and 625.

[00266] In some embodiments, a heavy chain-encoding polynucleotide is characterized by at least 85%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% nucleic acid sequence identity with a polynucleotide of SEQ ID NO: 16, 48, 80, or 611. In some embodiments, a polynucleotide encoding a light chain is characterized by at least 85%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% nucleic acid sequence identity with a polynucleotide under SEQ ID NO: 32, 64, 96 or 627.

[00267] The polynucleotides of the present invention can encode only the sequence of the variable region of an anti-CCR7 antibody. They can also encode both the variable region and the constant region of an antibody. Some of the polynucleotide sequences encode a polypeptide that contains both the heavy chain and light chain variable regions of one of the exemplary mouse anti-CCR7 antibodies. Some other polynucleotides encode two polypeptide segments that are largely identical to the heavy chain and light chain variable regions, respectively, of one of the mouse antibodies.

[00268] Polynucleotide sequences can be generated by de novo solid phase DNA synthesis or by PCR mutagenesis of an existing sequence (eg, those described in the Examples below) encoding an anti-CCR7 antibody or binding fragment thereof. Direct chemical synthesis of nucleic acids can be carried out using methods known in the art, such as the phosphotriester method from Narang et al., Meth. Enzymol. 68:90, 1979; the phosphodiester method from Brown et al., Meth. Enzymol. 68:109, 1979; diethylphosphoramidite method from Beaucage et al., Tetra Lett., 22:1859, 1981; and the solid support method of US Pat. No. 4,458,066. Introduction of mutations into a polynucleotide sequence by PCR can be performed as described, for example, in PCR Technology: Principles and Applications for DNA Amplification, HA Erlich (Ed.), Freeman Press, NY , NY, 1992; PCR Protocols: A Guide to Methods and Applications, Innis et al. (Ed.), Academic Press, San Diego, CA, 1990; Mattila et al., Nucleic Acids Res. 19:967, 1991; and Eckert et al., PCR Methods and Applications 1:17, 1991.

[00269] Also provided herein are expression vectors and host cells for producing an anti-CCR7 antibody as described above. Various expression vectors can be used to express polynucleotides encoding chains or binding fragments of an anti-CCR7 antibody. Both viral and non-viral expression vectors can be used to produce antibodies in a mammalian host cell. Non-viral vectors and systems include plasmids, episomal vectors, typically with an expression cassette for protein or RNA expression, and artificial human chromosomes (see, for example, Harrington et al., Nat Genet. 15:345, 1997). For example, non-viral vectors useful for expressing CCR7-binding polynucleotides and polypeptides in mammalian (eg, human) cells include pThioHis A, B, and C, pcDNA ^™ 3.1/His, pEBVHis A, B, and C (Invitrogen, San Diego , Calif.), MPSV vectors, and numerous other vectors known in the art for expressing other proteins. Useful viral vectors include retrovirus, adenovirus, adeno-associated virus, herpesvirus, SV40, papillomavirus, Epstein-Barr HBP, vaccinia virus, and Semliki forest virus (SFV) vectors. See Brent et al., supra; Smith, Anna. Rev. microbiol. 49:807, 1995; and Rosenfeld et al., Cell 68:143, 1992.

[00270] The choice of expression vector depends on the intended host cells in which the vector is to be expressed. Typically, expression vectors contain a promoter and other regulatory sequences (eg, enhancers) that are operably linked to polynucleotides encoding an anti-CCR7 antibody chain or fragment. In some embodiments, an inducible promoter is used to prevent expression of the inserted sequences under conditions other than inducing. Inducible promoters include, for example, the arabinose, lacZ, metallothionein or heat shock protein promoter. Cultures of transformed organisms can be propagated under non-inducing conditions without shifting the population towards coding sequences whose expression products are better tolerated by host cells. For efficient expression of an anti-CCR7 antibody chain or fragment, other regulatory elements may also be needed or required in addition to promoters. Such elements typically include the translation initiation codon ATG and an adjacent ribosome binding site or other sequences. In addition, expression efficiency can be increased by including enhancers appropriate to the cell system used (see, for example, Scharf et al., Results Probl. Cell Differ. 20:125, 1994; and Bittner et al., Meth. Enzymol., 153:516, 1987). For example, an SV40 enhancer or a CMV enhancer can be used to increase expression in mammalian host cells.

[00271] Expression vectors can also be provided with a secretion signal sequence to form a fusion protein with polypeptides encoded by the inserted anti-CCR7 antibody sequences. More commonly, the inserted anti-CCR7 antibody sequences are fused to signal sequences prior to inclusion in the vector. The vectors to be used to obtain sequences encoding the light and heavy chain variable domains of an anti-CCR7 antibody sometimes also encode constant regions or portions thereof. Such vectors allow the expression of variable regions as fusion proteins with constant regions, thereby leading to the production of intact antibodies or fragments thereof. Typically, such constant regions are human.

[00272] Host cells carrying and expressing anti-CCR7 antibody chains can be either prokaryotic or eukaryotic. E. coli is one prokaryotic host useful for cloning and expression of the polynucleotides of the present invention. Other suitable microbial hosts include bacilli such as Bacillus subtilis and other enterobacteria such as Salmonella, Serratia and various Pseudomonas species. For these prokaryotic hosts, expression vectors can also be created, which typically contain expression control sequences compatible with the host cell (eg, origin of replication). In addition, any number of various well-known promoters will be present, such as the lactose promoter system, the tryptophan (trp) promoter system, the beta-lactamase promoter system, or the lambda phage promoter system. Typically, promoters drive expression, optionally with an operator sequence, and have ribosome binding site sequences and the like. for the initiation and completion of transcription and translation. Other microorganisms such as yeast can also be used to express the CCR7 binding polypeptides of the present invention. It is also possible to use insect cells in combination with baculovirus vectors.

[00273] In some preferred embodiments, mammalian host cells are used to express and produce the CCR7 binding polypeptides of the present invention. For example, they may be either a hybridoma cell line expressing endogenous immunoglobulin genes (e.g., hybridoma clones from myeloma cells as described in the Examples) or a mammalian cell line carrying an exogenous expression vector (e.g., SP2/0 myeloma cells as described in as an example below). They cover any normal mortal or normal or abnormal immortal animal or human cells. For example, a number of suitable host cell lines capable of secreting intact immunoglobulins have been developed, including CHO cell lines, various Cos cell lines, HeLa cells, myeloma cell lines, transformed B cells, and hybridomas. The use of tissue culture of mammalian cells for the expression of polypeptides is discussed in general terms in, for example, Winnacker, From Genes to Clones, VCH Publishers, NY, NY, 1987. Expression vectors for mammalian host cells may include sequences to direct expression , such as the origin of replication, promoter and enhancer (see, for example, Queen et al., Immunol. Rev. 89:49-68, 1986), and necessary sites carrying information for processing, such as ribosome binding sites, sites RNA splicing, polyadenylation sites, and transcription terminator sequences. Such expression vectors typically contain promoters derived from mammalian genes or from mammalian viruses. Suitable promoters can be constitutive, cell type specific, developmental stage specific, and/or modulated or regulated. Useful promoters include, but are not limited to, the metallothionein promoter, adenovirus constitutive large late promoter, dexamethasone-inducible MMTV promoter, SV40 promoter, MRP polIII promoter, MPSV constitutive promoter, tetracycline-inducible CMV promoter (such as the human CMV immediate-early promoter), constitutive CMV promoter, and combinations promoter-enhancer, known from the prior art.

[00274] Methods for introducing expression vectors containing polynucleotide sequences of interest vary depending on the type of host cell. For example, calcium chloride transfection is typically used on prokaryotic cells, while calcium phosphate treatment or electroporation can be used on other host cells (see outlined in Sambrook et al., supra). A Laboratory Manual, 4th ^ed .). Other methods include, for example, electroporation, calcium phosphate treatment, liposome-mediated transformation, injection and microinjection, ballistic methods, virosomes, immunoliposomes, polycation:nucleic acid conjugates, deproteinized DNA, artificial virions, fusion with the structural protein VP22 of the herpes virus (Elliot and O'Hare, Cell 88:223, 1997), agent-enhanced DNA uptake and ex vivo transduction. To ensure long-term production of recombinant proteins in high yield, stable expression will often be required. For example, cell lines that stably express anti-CCR7 antibody chains or binding fragments can be generated using expression vectors of the present invention that contain viral origins or endogenous expression elements and a selectable marker gene. After the introduction of the vector, cells can be left to grow for 1-2 days in enriched medium before they are transferred to selective medium. The purpose of the selectable marker is to confer resistance to selection factors and its presence allows cells that successfully express the introduced sequences to grow in selective media. Proliferation of resistant stably transfected cells can be achieved using tissue culture techniques appropriate to the cell type.

Therapeutic routes

[00275] The antibodies, antibody fragments (e.g., antigen-binding fragments), and antibody-drug conjugates of the present invention are useful in a variety of applications, including, without limitation, the treatment or prevention of cancer, such as types of solid cancer or hematological malignancies. In some embodiments, the antibodies, antibody fragments (eg, antigen-binding fragments), and antibody-drug conjugates of the present invention are useful for inhibiting tumor growth, inducing differentiation, reducing tumor volume, and/or reducing tumor tumorigenicity. The methods of application may be in vitro , ex vivo or in vivo methods.

[00276] In one aspect, antibodies, antibody fragments (eg, antigen-binding fragments), and antibody-drug conjugates of the present invention are useful for detecting the presence of CCR7 in a biological sample. As used herein, the term "performing detection" encompasses quantitative or qualitative detection. In some embodiments, the biological sample contains a cell or tissue. In some embodiments, such tissues include normal and/or cancerous tissues that express CCR7 at higher levels than other tissues.

[00277] In one aspect of the present invention, a method is provided for detecting the presence of CCR7 in a biological sample. In some embodiments, the method involves contacting a biological sample with an anti-CCR7 antibody under conditions that allow binding of the antibody to the antigen and determining if a complex is formed between the antibody and the antigen.

[00278] In one aspect, the present invention provides a method for diagnosing a disorder associated with overexpression of CCR7. In some embodiments, the method comprises contacting a test cell with an anti-CCR7 antibody; determining the expression level (either quantitatively or qualitatively) of CCR7 on a test cell by detecting the binding of an anti-CCR7 antibody to the CCR7 antigen and comparing the expression level of CCR7 in the test cell with the level of CCR7 expression on a control cell (e.g., a normal cell originating from the same tissue, as a test cell, or a cell that expresses CCR7 at levels comparable to those of a normal cell), where a higher level of CCR7 expression on the test cell compared to the control cell indicates the presence of a disorder associated with increased expression of CCR7. In some embodiments, the test cell is obtained from an individual suspected of having a disorder associated with overexpression of CCR7. In some embodiments, the implementation of the violation is a violation of cell proliferation, such as cancer or tumor. In some embodiments, the method includes measuring the copy number of the CCR7 gene in the test cell.

[00279] In some embodiments, a diagnostic or detection method, such as those described above, involves detecting binding of an anti-CCR7 antibody to CCR7 expressed on a cell surface or in a membrane preparation derived from a cell expressing CCR7 on its surface. An exemplary assay for detecting binding of an anti-CCR7 antibody to CCR7 expressed on a cell surface is the "FACS" assay.

[00280] Several other methods can be used to detect binding of an anti-CCR7 antibody to CCR7. Such methods include, without limitation, antigen binding assays that are well known in the art, such as Western blotting, radioimmunoassays, ELISA (enzyme-linked immunosorbent assay), sandwich immunoassays, immunoprecipitation assays, fluorescence immunoassays, protein immunoassays. A and immunohistochemistry (IHC).

[00281] In some embodiments, anti-CCR7 antibodies are labeled. Labels include, without limitation, labels or moieties that are detected directly (such as fluorescent, chromophoric, electron dense, chemiluminescent, and radioactive labels) as well as moieties, such as enzymes or ligands, that are detected indirectly, such as through an enzymatic reaction or molecular interaction.

[00282] In some embodiments, anti-CCR7 antibodies are immobilized on an insoluble matrix. Immobilization results in separation of the anti-CCR7 antibody from any CCR7 protein that remains free in solution. Traditionally, this is done either by rendering the anti-CCR7 antibody in an insoluble form prior to the assay procedure by adsorption to a water-insoluble matrix or surface (Bennich et al., US Pat. No. 3,720,760) or by covalent bonding (e.g., using glutaraldehyde crosslinking), or by converting the anti-CCR7 antibody into an insoluble form after complex formation between the anti-CCR7 antibody and the CCR7 protein, for example by immunoprecipitation.

[00283] Any of the above diagnostic or detection embodiments can be performed using an immunoconjugate of the present invention instead of or in addition to an anti-CCR7 antibody.

[00284] In one embodiment, the present invention provides a method of treating or preventing a disease, comprising administering to a patient antibodies, antibody fragments (eg, antigen-binding fragments), or antibody-drug conjugates of the present invention. The present invention also provides the use of antibodies, antibody fragments (eg, antigen-binding fragments), or antibody-drug conjugates of the present invention to treat or prevent a disease in a patient. In some embodiments, the present invention provides antibodies, antibody fragments (eg, antigen-binding fragments), or antibody-drug conjugates of the present invention for use in the treatment or prevention of a disease in a patient. In further embodiments, the present invention provides the use of antibodies, antibody fragments (eg, antigen-binding fragments), or antibody-drug conjugates of the present invention in the manufacture of a medicament for the treatment or prevention of a disease in a patient.

[00285] In some embodiments, the disease being treated with antibodies, antibody fragments (eg, antigen-binding fragments), and antibody-drug conjugates of the present invention is cancer. In some embodiments, the cancer is characterized by the presence of CCR7-expressing cells to which antibodies, antibody fragments (eg, antigen-binding fragments), and antibody-drug conjugates of the present invention bind. In some embodiments, the implementation of the cancer is characterized by an increase in the expression of CCR7 compared to the level of expression in a healthy patient. In some embodiments, CCR7 expression can be measured by an increase in CCR7 RNA. In other embodiments, the cancer is characterized by an increase in the copy number of CCR7 DNA. Those skilled in the art are aware of other methods for measuring or determining levels of CCR7 expression. Examples of diseases that can be treated and/or prevented include, without limitation, chronic lymphocytic leukemia (CLL), peripheral T-cell lymphomas (PTCLs) such as adult T-cell leukemia/lymphoma (ATLL) and anaplastic large cell lymphoma (ALCL) , non-Hodgkin's lymphoma (NHL) such as mantle cell lymphoma (MCL), Burkitt's lymphoma and diffuse large B cell lymphoma (DLBCL), gastric carcinoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, nasopharyngeal carcinoma (NPC ), esophageal cancer, colorectal carcinoma, pancreatic cancer, thyroid cancer, breast cancer, renal cell carcinoma, and cervical cancer.

[00286] The present invention provides methods for treating or preventing cancer comprising administering a therapeutically effective amount of antibodies, antibody fragments (eg, antigen-binding fragments), or antibody-drug conjugates of the present invention. In some embodiments, the cancer is a solid cancer. In some embodiments, the subject is a human. In some embodiments, the cancer is resistant cancer and/or recurrent cancer. In certain aspects, for example, resistant cancer is resistant to tyrosine kinase inhibitors, EGFR inhibitors, Her2 inhibitors, Her3 inhibitors, IGFR inhibitors, and Met inhibitors.

[00287] In some embodiments, the present invention provides methods for inhibiting tumor growth, comprising administering to a subject a therapeutically effective amount of antibodies, antibody fragments (eg, antigen-binding fragments), or antibody-drug conjugates of the present invention. In some embodiments, the subject is a human. In some embodiments, the subject has a tumor or has had a tumor removed. In some embodiments, the tumor is resistant to other tyrosine kinase inhibitors, including, but not limited to, EGFR inhibitors, Her2 inhibitors, Her3 inhibitors, IGFR inhibitors, and Met inhibitors.

[00288] In some embodiments, the implementation of the tumor expresses CCR7, which binds antibodies to CCR7. In some embodiments, the tumor overexpresses human CCR7. In some embodiments, the tumor is characterized by an increased copy number of the CCR7 gene.

[00289] The present invention also provides methods for selecting patients for treatment with antibodies, antibody fragments (e.g., antigen-binding fragments), or antibody-drug conjugates of the present invention, comprising administering a therapeutically effective amount of said antibodies, antibody fragments (e.g., antigen-binding fragments), or antibody-drug conjugates. In certain aspects, the method involves the selection of patients with cancer resistant to tyrosine kinase inhibitors. In certain aspects, tyrosine kinase inhibitor-resistant cancers are expected to be resistant to EGFR inhibitors, Her2 inhibitors, Her3 inhibitors, IGFR inhibitors, and/or Met inhibitors. In certain aspects, it is assumed that resistant cancer is a cancer that is resistant to Her2. In certain aspects, the cancer is assumed to be de novo resistant cancer, and in still other aspects, the cancer is assumed to be recurrent cancer. In certain aspects of the present invention, the methods involve selecting a patient with de novo resistant or recurrent cancer and measuring CCR7 expression. It is contemplated that, in certain aspects, the recurrent cancer or tumor was not originally a CCR7 expressing cancer or tumor, but becomes a CCR7 positive cancer, which is a tyrosine kinase resistant or recurrent cancer or tumor, following treatment with tyrosine kinase inhibitors.

[00290] In the case of treating or preventing diseases, the appropriate dose of antibodies, antibody fragments (e.g., antigen-binding fragments), or antibody-drug conjugates of the present invention depends on various factors such as the type of disease being treated, the severity and course of the disease, the response disease, previous therapy, the patient's clinical history, etc. The antibody or agent may be administered once or throughout a course of treatment lasting from a few days to several months, or until a cure is achieved, or until an amelioration of the disease state is achieved (for example, a reduction in the size tumors). Optimal dosing regimens can be calculated based on measurements of drug accumulation in the patient's body and will vary depending on the relative potency of individual antibodies, antibody fragments (eg, antigen-binding fragment), or antibody-drug conjugates. The attending physician can estimate the frequency of repetition of the dose based on the measured retention time and concentrations of the drug in body fluids or tissues.

Combination Therapy

[00291] In certain instances, the antibody, antibody fragment (e.g., antigen-binding fragment), or antibody-drug conjugate of the present invention is combined with other therapeutic agents, such as other anti-cancer agents, anti-allergy agents, anti-nausea (or anti-emetics), pain relievers agents, cytoprotective agents and combinations thereof.

[00292] In one embodiment, an antibody, antibody fragment (e.g., antigen-binding fragment), or antibody-drug conjugate of the present invention is combined in a pharmaceutical combination formulation with a second compound having anti-cancer properties, or a dosing schedule provides for combination therapy with it. The second compound of a pharmaceutical combination formulation or dosing regimen may have activities that complement the antibody or immunoconjugate of the combination such that they do not adversely affect each other. For example, an antibody, antibody fragment (e.g., antigen-binding fragment), or antibody-drug conjugate of the present invention may be administered in combination with, without limitation, a chemotherapeutic agent, a tyrosine kinase inhibitor, an inhibitor of the CCR7 downstream signaling pathway, IAP inhibitors, Bcl2 inhibitors, Mcl1 and other CCR7 inhibitors.

[00293] As used herein, the term "pharmaceutical combination" refers to either a fixed combination in the form of a dosage unit, or a non-fixed combination or set of parts for combined administration, where two or more therapeutic agents can be administered independently at the same time or separately within time intervals, in particular when such time intervals provide the opportunity to demonstrate the cooperative, for example synergistic, effect of the partners in the combination.

[00294] The term "combination therapy" refers to the administration of two or more therapeutic agents for the treatment or prevention of a therapeutic condition or disorder described in the present invention. Such administration encompasses the co-administration of these therapeutic agents substantially simultaneously, for example in a single capsule having a fixed ratio of active ingredients. Alternatively, such administration encompasses co-administration in multiple or separate containers (eg capsules, powders and liquids) for each active ingredient. Powders and/or liquids may be reconstituted or diluted to the desired dose prior to administration. In addition, such administration also encompasses the use of each type of therapeutic agent in a sequential manner, either at approximately the same time or at different times. In any event, the treatment regimen will provide the beneficial effects of the combination of drugs in the treatment or prevention of the conditions or disorders described herein.

[00295] Combination therapy can provide "synergy" and be "synergistic", ie. the effect achieved when the active ingredients are applied together is greater than the sum of the effects that result from the use of the compounds alone. A synergistic effect can be achieved when the active ingredients are: (1) formulated together and administered or delivered simultaneously as a single dose combination formulation; (2) are delivered sequentially or in parallel as separate trains, or (3) some other scheme is used. When delivered as interleaved therapy, a synergistic effect can be achieved when the compounds are administered or delivered one after the other, for example by different injections in separate syringes. In general, during interleaved therapy, an effective dose of each active ingredient is administered one after the other, i. series, while in combination therapy, an effective dose of two or more active ingredients is administered together.

[00296] Common chemotherapeutic agents considered for use in combination therapies include anastrozole ( ^Arimidex® ), bicalutamide ( ^Casodex® ), bleomycin sulfate (Blenoxane®), busulfan ( ^Myleran® ), ^busulfan injectable ( ^Busulfex® ), capecitabine (Xeloda ^® ), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin (Paraplatin ^® ), carmustine (BiCNU ^® ), chlorambucil (Leukeran ^® ), cisplatin (Platinol ^® ), cladribine (Leustatin ^® ), cyclophosphamide ( Cytoxan ^® or Neosar ^® ), cytarabine, cytosine arabinoside (Cytosar-U ^® ), cytarabine liposomal injection (DepoCyt ^® ), dacarbazine (DTIC-Dome ^® ), dactinomycin (Actinomycin D, Cosmegan), daunorubicin hydrochloride (Cerubidine ^® ) , daunorubicin citrate liposomal injection (DaunoXome ^® ), dexamethasone, docetaxel (Taxotere ^® ), doxorubicin hydrochloride (Adriamycin ^® , Rubex ^® ), etoposide (Vepesid ^® ), fludarabine phosphate (Fludara ^® ), 5-fluorouracil (Adrucil ^® , Efudex ^® ), flutamide (Eulexin ^® ), tezacitibine, gemcitabine (difluorodeoxycytidine), hydroxyurea (Hydrea ^® ), idarubicin (Idamycin ^® ), ifosfamide (IFEX ^® ), irinotecan (Camptosar ^® ), L-asparaginase (ELSPAR ^® ), calcium leucovorin, melphalan (Alkeran ^® ), 6-mercaptopurine (Purinethol ^® ), methotrexate (Folex ^® ), mitoxantrone (Novantrone ^® ), mylotarg, paclitaxel (Taxol ^® ), phoenix (yttrium 90/MX-DTPA), pentostatin, polyferospan 20 with carmustine implant (Gliadel ^® ), tamoxifen citrate (Nolvadex ^® ), teniposide (Vumon ^® ), 6-thioguanine, thiotepa, tirapazamine (Tirazone ^® ), topotecan hydrochloride injection (Hycamptin ^® ), vinblastine (Velban ^® ), vincristine (Oncovin ^® ), and vinorelbine (Navelbine ^® ), as well as pemetrexed.

[00297] In one aspect, the present invention provides a method of treating or preventing cancer by administering to a subject in need thereof an antibody drug conjugate of the present invention in combination with one or more tyrosine kinase inhibitors, including but not limited to BTK inhibitors, EGFR inhibitors, Her2 inhibitors, Her3 inhibitors, IGFR inhibitors and Met inhibitors.

[00298] For example, tyrosine kinase inhibitors include, without limitation, ibrutinib (PCI-32765); erlotinib hydrochloride (Tarceva®); linifanib (N-[4-(3-amino-1H-indazol-4-yl)phenyl]-N'-(2-fluoro-5-methylphenyl)urea, also known as ABT 869 available from Genentech); sunitinib malate (Sutent®); bosutinib (4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methylpiperazin-1-yl)propoxy]quinolin-3-carbonitrile, also known as SKI- 606 and described in US patent No. 6780996); dasatinib (Sprycel®); pazopanib (Votrient®); sorafenib (Nexavar®); zactima (ZD6474); and imatinib or imatinib mesylate (Gilvec® and Gleevec®).

[00299] Epidermal growth factor receptor receptor (EGFR) inhibitors include, without limitation, erlotinib hydrochloride (Tarceva®), gefitinib (Iressa®); N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[[(3''S'')-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4(dimethylamino)- 2-butenamide, Tovok®); vandetanib (Caprelsa®); lapatinib (Tykerb®); (3R,4R)-4-amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3 -ol (BMS690514); canertinib dihydrochloride (CI-1033); 6-[4-[(4-ethyl-1-piperazinyl)methyl]phenyl]-N-[(1R)-1-phenylethyl]- 7H-pyrrolo[2,3-d]pyrimidin-4-amine (AEE788, CAS 497839-62-0); mubritinib (TAK165); pelitinib (EKB569); afatinib (BIBW2992); neratinib (HKI-272); N-[4-[[1-[(3-fluorophenyl)methyl]-1H-indazol-5-yl]amino]-5-methylpyrrolo[2,1-f][1,2,4]triazin-6- yl]-carbamic acid, (3S)-3-morpholinylmethyl ester (BMS599626); N-(3,4-dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4-quinazolinamine (XL647, CAS 781613-23-8) and 4-[4-[[(1R)-1-phenylethyl]amino]-7H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol (PKI166, CAS 187724-61-4).

[00300] Antibodies that bind EGFR include, without limitation, cetuximab (Erbitux®); panitumumab (Vectibix®); matuzumab (EMD-72000); nimotuzumab (hR3); zalutumumab; TheraCIM h-R3; MDX0447 (CAS 339151-96-1) and ch806 (mAb-806, CAS 946414-09-1).

[00301] Human epidermal growth factor 2 receptor (Her2 receptor) inhibitors (also known as Neu, ErbB-2, CD340, or p185) include, but are not limited to, trastuzumab (Herceptin®); pertuzumab (Omnitarg®); trastuzumab emtansine (Kadcyla®); neratinib (HKI-272, (2E)-N-[4-[[3-chloro-4-[(pyridin-2-yl)methoxy]phenyl]amino]-3-cyano-7-ethoxyquinolin-6-yl] -4-(dimethylamino)but-2-enamide and described in PCT Publication No. WO 05/028443); lapatinib or lapatinib ditosylate (Tykerb®); (3R,4R)-4-amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3 -ol (BMS690514); (2E)-N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4-(dimethylamino)- 2-butenamide (BIBW-2992, CAS 850140-72-6); N-[4-[[1-[(3-fluorophenyl)methyl]-1H-indazol-5-yl]amino]-5-methylpyrrolo[2,1-f][1,2,4]triazin-6- yl]-carbamic acid, (3S)-3-morpholinylmethyl ester (BMS 599626, CAS 714971-09-2); canertinib dihydrochloride (PD183805 or CI-1033) and N-(3,4-dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrole- 5-yl]methoxy]-4-quinazolinamine (XL647, CAS 781613-23-8).

[00302] Her3 inhibitors include, without limitation, LJM716, MM-121, AMG-888, RG7116, REGN-1400, AV-203, MP-RM-1, MM-111, and MEHD-7945A.

[00303] Met inhibitors include, without limitation, cabozatinib (XL184, CAS 849217-68-1); foretinib (GSK1363089, formerly XL880, CAS 849217-64-7); tivantinib (ARQ197, CAS 1000873-98-2); 1-(2-hydroxy-2-methylpropyl) -N- (5-(7-methoxyquinolin-4-yloxy)pyridin-2-yl)-5-methyl-3-oxo-2-phenyl-2,3-dihydro -1H-pyrazole-4-carboxamide (AMG 458); crizotinib (Xalkori®, PF-02341066); (3Z)-5-(2,3-dihydro-1H-indol-1-ylsulfonyl)-3-({3,5-dimethyl-4-[(4-methylpiperazin-1-yl)carbonyl]-1H-pyrrole -2-yl}methylene)-1,3-dihydro-2H-indol-2-one (SU11271); (3Z)-N-(3-chlorophenyl)-3-({3,5-dimethyl-4-[(4-methylpiperazin-1-yl)carbonyl]-1H-pyrrol-2-yl}methylene)-N- methyl-2-oxoindoline-5-sulfonamide (SU11274); (3Z)-N-(3-chlorophenyl)-3-{[3,5-dimethyl-4-(3-morpholin-4-ylpropyl)-1H-pyrrol-2-yl]methylene}-N-methyl-2 -oxoindoline-5-sulfonamide (SU11606); 6-[difluoro[6-(1-methyl-1H-pyrazol-4-yl)-1,2,4-triazolo[4,3-b]pyridazin-3-yl]methyl]-quinoline (JNJ38877605, CAS 943540 -75-8); 2-[4-[1-(quinolin-6-ylmethyl)-1H-[1,2,3]triazolo[4,5-b]pyrazin-6-yl]-1H-pyrazol-1-yl]ethanol ( PF04217903, CAS 956905-27-4); N-((2R)-1,4-dioxan-2-ylmethyl)-N-methyl-N'-[3-(1-methyl-1H-pyrazol-4-yl)-5-oxo-5H-benzo[ 4,5]cyclohepta[1,2-b]pyridin-7-yl]sulfamide (MK2461, CAS 917879-39-1); 6-[[6-(1-methyl-1H-pyrazol-4-yl)-1,2,4-triazolo[4,3- b ]pyridazin-3-yl]thio]-quinoline (SGX523, CAS 1022150- 57-7) and (3Z)-5-[[(2,6-dichlorophenyl)methyl]sulfonyl]-3-[[3,5-dimethyl-4-[[(2R)-2-(1-pyrrolidinylmethyl) -1-pyrrolidinyl]carbonyl]-1H-pyrrol-2-yl]methylene]-1,3-dihydro-2H-indol-2-one (PHA665752, CAS 477575-56-7).

[00304] IGF1R inhibitors include, without limitation, BMS-754807, XL-228, OSI-906, GSK0904529A, A-928605, AXL1717, KW-2450, MK0646, AMG479, IMCA12, MEDI-573 and BI836845. For a review, see, for example, Yee, JNCI, 104; 975 (2012).

[00305] In another aspect, the present invention provides a method of treating or preventing cancer by administering to a subject in need thereof an antibody drug conjugate of the present invention in combination with one or more CCR7 downstream signaling pathway inhibitors, including but not limited to β-arrestin inhibitors, GRK inhibitors, MAPK inhibitors, PI3K inhibitors, JAK inhibitors, etc.

[00306] For example, phosphoinositide 3-kinase (PI3K) inhibitors include, but are not limited to, idelalisib (Zydelig, GS-1101, Cal-101), 4-[2-(1H-indazol-4-yl)-6-[[4- (methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as GDC 0941 and described in PCT Publication Nos. WO 09/036082 and WO 09/055730) ; 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile ( also known as BEZ 235 or NVP-BEZ 235 and described in PCT Publication No. WO 06/122806); 4-(trifluoromethyl)-5-(2,6-dimorpholinopyrimidin-4-yl)pyridine-2-amine (also known as BKM120 or NVP-BKM120 and described in PCT Publication No. WO2007/084786); tosacertib (VX680 or MK-0457, CAS 639089-54-6); (5Z)-5-[[4-(4-pyridinyl)-6-quinolinyl]methylene]-2,4-thiazolidinedione (GSK1059615, CAS 958852-01-2); (1E,4S,4aR,5R,6aS,9aR)-5-(acetyloxy)-1-[(di-2-propenylamino)methylene]-4,4a,5,6,6a,8,9,9a-octahydro -11-hydroxy-4-(methoxymethyl)-4a,6a-dimethyl-cyclopenta[5,6]naphtho[1,2-c]pyran-2,7,10(1H)-trione (PX866, CAS 502632-66 -8) and 8-phenyl-2-(morpholin-4-yl)-chromen-4-one (LY294002, CAS 154447-36-6).

[00307] In yet another aspect, the present invention provides a method of treating or preventing cancer by administering to a subject in need thereof an antibody drug conjugate of the present invention in combination with one or more pro-apoptotic agents, including but not limited to IAP inhibitors, Bcl2 inhibitors , MCl1 inhibitors, Trail agents, Chk inhibitors.

[00308] For example, IAP inhibitors include, without limitation, LCL161, GDC-0917, AEG-35156, AT406, and TL32711. Other examples of IAP inhibitors include, without limitation, those disclosed in WO04/005284, WO 04/007529, WO05/097791, WO 05/069894, WO 05/069888, WO 05/094818, US2006/0014700, US2006/0025347, WO 063/06900 WO 06/010118, WO 06/017295 and WO08/134679, all of which are incorporated herein by reference.

[00309] BCL-2 inhibitors include, without limitation, venetoclax (also known as GDC-0199, ABT-199, RG7601); 4-[4-[[2-(4-chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1-piperazinyl]-N-[[4-[[(1R)-3 -(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3-[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide (also known as ABT-263 and described in PCT Publication No. WO 09/ 155386); tetracarcin A; antimycin; gossypol ((-)BL-193); obatoclax; ethyl 2-amino-6-cyclopentyl-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromone-3-carboxylate (HA14-1); oblimersen (G3139, Genasense®); Bak BH3 peptide; (-)-gossypol-acetic acid (AT-101); 4-[4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]-1-piperazinyl]- N -[[4-[[(1 R )-3-(dimethylamino) -1-[(phenylthio)methyl]propyl]amino]-3-nitrophenyl]sulfonyl]-benzamide (ABT-737, CAS 852808-04-9) and navitoclax (ABT-263, CAS 923564-51-6).

[00310] Pro-apoptotic receptor (PARA) agonists, including DR4 (TRAILR1) and DR5 (TRAILR2), include, without limitation, dulanermin (AMG-951, RhApo2L/TRAIL); mapatumumab (HRS-ETR1, CAS 658052-09-6); lexatumumab (HGS-ETR2, CAS 845816-02-6); apomab (Apomab®); conatumumab (AMG655, CAS 896731-82-1) and tigatuzumab (CS1008, CAS 946415-34-5, available from Daiichi Sankyo).

[00311] Checkpoint kinase (CHK) inhibitors include, without limitation, 7-hydroxystaurosporine (UCN-01); 6-bromo-3-(1-methyl-1H-pyrazol-4-yl)-5-(3R)-3-piperidinyl-pyrazolo[1,5- a ]pyrimidine-7-amine (SCH900776, CAS 891494-63 -6); 5-(3-fluorophenyl)-3-ureidothiophene-2-carboxylic acid N-[(S)-piperidin-3-yl]amide (AZD7762, CAS 860352-01-8); 4-[((3S)-1-azabicyclo[2.2.2]oct-3-yl)amino]-3-(1H-benzimidazol-2-yl)-6-chloroquinolin-2(1H)-one (CHIR 124 , CAS 405168-58-3); 7-aminodactinomycin (7-AAD), isogranululathymide, debromohymenialdizine; N-[5-bromo-4-methyl-2-[(2S)-2-morpholinylmethoxy]phenyl]-N'-(5-methyl-2-pyrazinyl)urea (LY2603618, CAS 911222-45-2); sulforaphane (CAS 4478-93-7, 4-methylsulfinyl butyl isothiocyanate); 9,10,11,12-tetrahydro-9,12-epoxy-1H-diindolo[1,2,3- fg :3',2',1'- kl ]pyrrolo[3,4- i ][1, 6]benzodiazocin-1,3(2H)-dione (SB-218078, CAS 135897-06-2) and TAT-S216A (YGRKKRRQRRRLYRSPAMPENL (SEQ ID NO: 629)) and CBP501 ((d-Bpa)sws( d-Phe-F5)(d-Cha)rrrqrr).

[00312] In a further embodiment, the present invention provides a method of treating or preventing cancer by administering to a subject in need thereof an antibody drug conjugate of the present invention in combination with one or more immunomodulators (e.g., one or more of a costimulatory molecule activator or inhibitor immune checkpoint molecules).

[00313] In some embodiments, the immunomodulator is a costimulatory molecule activator. In one embodiment, the co-stimulatory molecule agonist is selected from an agonist (e.g., agonist antibody or antigen-binding fragment thereof, or soluble fusion protein) OX40, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278) , 4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, STING or CD83 ligand.

[00314] In some embodiments, the immunomodulator is an inhibitor of an immune checkpoint molecule. In one embodiment, the immunomodulator is an inhibitor of PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR-beta. In one embodiment, the immune checkpoint molecule inhibitor inhibits PD-1, PD-L1, LAG-3, TIM-3, or CTLA4, or any combination thereof. The term "inhibition" or "inhibitor" refers to the reduction of a certain parameter, such as the activity of a given molecule, such as a checkpoint inhibitor. For example, the term refers to inhibition of an activity, eg, PD-1 or PD-L1 activity, by at least 5%, 10%, 20%, 30%, 40%, 50%, or more. Thus, inhibition need not be 100%.

[00315] Inhibition of an inhibitory molecule can occur at the DNA, RNA, or protein level. In some embodiments, an inhibitory nucleic acid (eg, dsRNA, siRNA, or shRNA) may be used to suppress the expression of an inhibitory molecule. In other embodiments, the inhibitor of the inhibitory signal is a polypeptide, such as a soluble ligand (eg, an Ig that binds PD-1 or an Ig that binds CTLA-4) or an antibody or antigen-binding fragment thereof, that binds to an inhibitory molecule; for example, an antibody or fragment thereof (also referred to herein as an "antibody molecule") that binds to PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR-beta or a combination thereof.

[00316] In one embodiment, the antibody molecule is a complete antibody or fragment thereof (eg, Fab, F(ab') ₂ , Fv, or single chain Fv fragment (scFv)). In other embodiments, the antibody molecule has a heavy chain constant region (Fc) selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE heavy chain constant regions; in particular selected, for example, from IgG1, IgG2, IgG3 and IgG4 heavy chain constant regions, more specifically IgG1 or IgG4 heavy chain constant region (eg human IgG1 or IgG4). In one embodiment, the heavy chain constant region is derived from human IgG1 or human IgG4. In one embodiment, the constant region is altered, such as mutated to modify properties of the antibody molecule (eg, increase or decrease one or more of Fc receptor binding, antibody glycosylation, cysteine residue number, effector cell function, or complement function).

[00317] In some embodiments, the antibody molecule is in the form of a bispecific or polyspecific antibody molecule. In one embodiment, the bispecific antibody molecule is characterized by a first binding specificity for PD-1 or PD-L1 and a second binding specificity, for example, a second binding specificity for TIM-3, LAG-3 or PD-L2. In one embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and TIM-3. In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and LAG-3. In another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L1. In yet another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L2. In another embodiment, the bispecific antibody molecule binds to TIM-3 and LAG-3. Any combination of the above molecules can be formulated into a polyspecific antibody molecule, e.g. a trispecific antibody that provides a first binding specificity for PD-1 or PD-1, as well as a second and third binding specificity for two or more of TIM-3, LAG-3 or PD-L2.

[00318] In some embodiments, the immunomodulator is a PD-1 inhibitor, such as human PD-1. In another embodiment, the immunomodulator is a PD-L1 inhibitor, such as human PD-L1. In one embodiment, the PD-1 or PD-L1 inhibitor is an anti-PD-1 or PD-L1 antibody molecule. The PD-1 or PD-L1 inhibitor may be administered alone or in combination with other immunomodulators, for example in combination with a LAG-3, TIM-3 or CTLA4 inhibitor. In an exemplary embodiment, a PD-1 or PD-L1 inhibitor, eg, an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a LAG-3 inhibitor, eg, an anti-LAG-3 antibody molecule. In another embodiment, a PD-1 or PD-L1 inhibitor, eg, an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a TIM-3 inhibitor, eg, an anti-TIM-3 antibody molecule. In other embodiments, a PD-1 or PD-L1 inhibitor, e.g., an anti-PD-1 antibody molecule, is administered in combination with a LAG-3 inhibitor, e.g., an anti-LAG-3 antibody molecule, and a TIM-3 inhibitor, e.g., an anti-LAG-3 antibody molecule. to TIM-3. Other combinations of immunomodulators with a PD-1 inhibitor (eg, one or more of PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, and/or TGFR) are also contemplated within the scope of the present invention. Any of the antibody molecules known in the art or disclosed herein can be used in the above combination of checkpoint molecule inhibitors.

[00319] In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody selected from nivolumab, pembrolizumab, or pidilizumab. In some embodiments, the anti-PD-1 antibody is nivolumab. Alternative names for nivolumab include MDX-1106, MDX-1106-04, ONO-4538, or BMS-936558. In some embodiments, the anti-PD-1 antibody is nivolumab (CAS accession number: 946414-94-4). Nivolumab is a fully human IgG4 monoclonal antibody that specifically blocks PD1. Nivolumab (clone 5C4) and other human monoclonal antibodies that specifically bind to PD1 are disclosed in US Pat. No. 8,008,449 and PCT Publication No. WO2006/121168.

[00320] In other embodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab (trade name KEYTRUDA, formerly lambrolizumab, also known as Merck 3745, MK-3475, or SCH-900475) is a humanized IgG4 monoclonal antibody that binds to PD1. Pembrolizumab is disclosed, for example, in Hamid, O. et al . (2013) New England Journal of Medicine 369 (2): 134-44, PCT Publication No. WO2009/114335 and US Patent No. 8354509.

[00321] In some embodiments, the anti-PD-1 antibody is pidilizumab. Pidilizumab (CT-011; Cure Tech) is a humanized IgG1k monoclonal antibody that binds to PD1. Pidilizumab and other humanized anti-PD-1 monoclonal antibodies are disclosed in PCT Publication No. WO2009/101611. Other anti-PD1 antibodies are disclosed in US Patent No. 8609089, US Patent Application Publication No. 2010028330 and/or US Patent Application Publication No. 20120114649. Other anti-PD1 antibodies include AMP 514 (Amplimmune).

[00322] In some embodiments, the PD-1 inhibitor is PDR001 or any other anti-PD-1 antibody disclosed in WO2015/112900.

[00323] In some embodiments, the PD-1 inhibitor is an immunoadhesin (e.g., an immunoadhesin comprising an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region (e.g., the Fc region of an immunoglobulin sequence). B in some embodiments, the PD-1 inhibitor is AMP-224.

[00324] In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody inhibitor is selected from YW243.55.S70, MPDL3280A, MEDI-4736, or MDX-1105MSB-0010718C (also referred to as A09-246-2) disclosed in e.g. WO 2013/0179174, and has a sequence disclosed herein (or a sequence substantially identical or similar to it, for example, a sequence that has at least 85%, 90%, 95% or higher identity with the specified sequence).

[00325] In one embodiment, the PD-L1 inhibitor is MDX-1105. MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in PCT Publication No. WO2007/005874.

[00326] In one embodiment, the PD-L1 inhibitor is YW243.55.S70. Antibody YW243.55.S70 is an anti-PD-L1 antibody described in PCT Publication No. WO 2010/077634 (heavy and light chain variable region sequences shown under SEQ ID Nos: 20 and 21, respectively).

[00327] In one embodiment, the PD-L1 inhibitor is MDPL3280A (Genentech/Roche). MDPL3280A is an Fc-optimized human IgG1 monoclonal antibody that binds to PD-L1. MDPL3280A and other human anti-PD-L1 monoclonal antibodies are disclosed in US Patent No. 7,943,743 and US Patent Application Publication No. 20120039906.

[00328] In other embodiments, the PD-L2 inhibitor is AMP-224. AMP-224 is a soluble PD-L2-Fc fusion protein receptor that blocks the interaction between PD1 and B7-H1 (B7-DCIg; Amplimmune; e.g. disclosed in PCT Publication Nos. WO2010/027827 and WO2011/066342) .

[00329] In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody molecule. In one embodiment, the LAG-3 inhibitor is BMS-986016.

Pharmaceutical compositions

[00330] To prepare pharmaceutical or sterile compositions containing immunoconjugates, the immunoconjugates of the present invention are mixed with a pharmaceutically acceptable carrier or excipient. The compositions may additionally contain one or more other therapeutic agents that are suitable for the treatment or prevention of cancer expressing CCR7 (including, without limitation, chronic lymphocytic leukemia (CLL), peripheral T-cell lymphomas (PTCL), such as T-cell leukemia/ adult lymphoma (ATLL) and anaplastic large cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL) such as mantle cell lymphoma (MCL), Burkitt's lymphoma and diffuse large B-cell lymphoma (DLBCL), gastric carcinomas, non-small cell lung cancer, small cell lung cancer, head and neck cancer, nasopharyngeal carcinoma (NPC), esophageal cancer, colorectal carcinoma, pancreatic cancer, thyroid cancer, breast cancer, renal cell carcinoma, and cervical cancer).

[00331] Therapeutic and diagnostic formulations can be prepared by mixing with physiologically acceptable carriers, excipients, or stabilizers in the form of, for example, lyophilized powders, slurries, aqueous solutions, lotions, or suspensions (see, for example, Hardman et al., Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY, 2001; Gennaro, Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY, 2000; Avis, et al. ( eds.), Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY, 1993; Lieberman, et al. (eds.), Pharmaceutical Dosage Forms: tablets, Marcel Dekker, NY, 1990; Lieberman, et al. (eds.) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY, 1990; Weiner and Kotkoskie, Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY, 2000).

[00332] The choice of administration schedule for a therapeutic agent depends on several factors, including the rate of serum or tissue metabolism for the subject, the level of symptoms, the immunogenicity of the subject, and the availability of target cells in the biological matrix. In some embodiments, the implementation schedule maximizes the amount of therapeutic agent delivered to the patient, in accordance with an acceptable level of side effects. Accordingly, the amount of biological agent delivered depends in part on the particular subject and the severity of the condition being treated. Guidelines for selecting appropriate doses of antibodies, cytokines and small molecules are available (see e.g. Wawrzynczak, Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK, 1996; Kresina (ed.), Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY, 1991; Bach (ed.), Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, Marcel Dekker, New York, NY, 1993; Baert et al., New Engl. J. Med. 348:601- 608, 2003; Milgrom et al., New Engl. J. Med. 341:1966-1973, 1999; Slamon et al., New Engl. J. Med. 344:783-792, 2001; Beniaminovitz et al., New Engl. J. Med. 342:613-619, 2000; Ghosh et al., New Engl. J. Med. 348:24-32, 2003; Lipsky et al., New Engl. J. Med. 343:1594- 1602, 2000).

[00333] Determination of an appropriate dose is performed by the attending physician, for example, using parameters or factors that are known or expected in the art to affect treatment or prevention or are predicted to affect treatment or prevention. Typically, dosing is started at a slightly less than optimal dose and thereafter increased in small increments until the desired or optimal effect is achieved in comparison to any negative side effects. Important diagnostic indicators include, for example, indicators that are symptoms of inflammation, or the level of inflammatory cytokines produced.

[00334] Actual dosage levels of active ingredients in the pharmaceutical compositions of the present invention may vary in order to obtain an amount of active ingredient that is effective to achieve the desired therapeutic response in a particular patient, effective in relation to the composition and method of administration, while not toxic to patient. The selected dose level will depend on a number of pharmacokinetic factors, including the potency of the particular compositions of the present invention or ester, salt or amide thereof used, route of administration, time of administration, rate of elimination of the particular compound used, duration of treatment, other drugs, compounds and /or materials used in combination with the specific compositions used, age, sex, body weight, condition, general health and history of the patient to be treated, and similar factors known in the field of medical technology.

[00335] Compositions containing antibodies or fragments thereof of the present invention can be administered by continuous infusion or by doses at intervals of, for example, one day, one week, or administered 1-7 times a week, once every two weeks , once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, or once every eight weeks. Doses may be administered intravenously, subcutaneously, topically, orally, nasally, rectally, intramuscularly, intracerebral, or by inhalation. A specific dosing protocol is one that provides for the maximum dose or frequency of dosing to avoid significant unwanted side effects.

[00336] For the immunoconjugates of the present invention, the dose administered to a patient may be from 0.0001 mg/kg to 100 mg/kg of the patient's body weight. The dose can be from 0.0001 mg/kg to 30 mg/kg, from 0.0001 mg/kg to 20 mg/kg, from 0.0001 mg/kg and 10 mg/kg, from 0.0001 mg/kg up to 5 mg/kg, from 0.0001 to 2 mg/kg, from 0.0001 to 1 mg/kg, from 0.0001 mg/kg to 0.75 mg/kg, from 0.0001 mg/kg to 0 .5 mg/kg, 0.0001 mg/kg to 0.25 mg/kg, 0.0001 to 0.15 mg/kg, 0.0001 to 0.10 mg/kg, 0.001 to 0, 5 mg/kg, 0.01 to 0.25 mg/kg, or 0.01 to 0.10 mg/kg of the patient's body weight. The dose of antibodies or fragments thereof of the present invention can be calculated using the patient's weight in kilograms (kg) multiplied by the dose to be administered in mg/kg.

[00337] Doses of the immunoconjugates of the present invention may be repeated and administrations may be separated by less than 1 day, at least 1 day, 2 days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months, 4 months, 5 months, or at least 6 months. In some embodiments, the immunoconjugates of the present invention may be given twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, or less frequently. In a specific embodiment, doses of the immunoconjugates of the present invention are administered again every 2 weeks.

[00338] The effective amount for a particular patient may vary depending on such factors as the condition being treated, the general health of the patient, the method, route and dose of administration, and the severity of side effects (see, for example, Maynard et al., A Handbook of SOPs for Good Clinical Practice, Interpharm Press, Boca Raton, Fla., 1996; Dent, Good Laboratory and Good Clinical Practice, Urch Publ., London, UK, 2001).

[00339] The route of administration may be, for example, topical or skin application, injection or infusion via subcutaneous, intravenous, intraperitoneal, intracerebral, intramuscular, intraocular, intraarterial, intraspinal, intralesional, or sustained release system or implant (see, e.g., Sidman et al., Biopolymers 22:547-556, 1983; Langer et al., J. Biomed. Mater. Res. 15:167-277, 1981; Langer, Chem. Tech. 12:98-105, 1982 ; Epstein et al., Proc. Natl. Acad. Sci. USA 82:3688-3692, 1985; Hwang et al., Proc. Natl. Acad. Sci. USA 77:4030-4034, 1980; U.S. Patent Nos. 6,350,466 and 6316024). If necessary, the composition may also contain a solubilizing agent and a local anesthetic such as lidocaine to relieve pain at the injection site, or both. In addition, inhalation administration may also be used, for example by using an inhaler or nebulizer and formulation with an aerosol agent. See, for example, US Pat. Nos. 6,019,968; 5,985,320; 5,985,309; 5,934,272; and PCT Publication Nos. WO 92/19244, WO 97/32572, WO 97/44013, WO 98/31346 and WO 99/66903, each of which is incorporated herein by reference in its entirety.

[00340] The composition of the present invention is also administered via one or more routes of administration using one or more of a variety of methods known in the art. As will be understood by a person skilled in the art, the route and/or method of administration will vary depending on the desired results. Selected routes of administration of the immunoconjugates of the present invention include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous routes of administration, injection into the spinal cord or other parenteral routes of administration, for example, by injection or infusion. Parenteral administration may be modes of administration other than enteral and topical administration, generally by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular , subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. Alternatively, the composition of the present invention may be administered via a route other than parenteral, such as topical, epidermal, or transmucosal routes, eg, intranasally, orally, vaginally, rectally, sublingually, or topically. In one embodiment, the immunoconjugates of the present invention are administered by infusion. In another embodiment, the immunoconjugates of the present invention are administered subcutaneously.

[00341] If the immunoconjugates of the present invention are administered in a controlled release or sustained release system, a pump may be used to effect the controlled or sustained release (see Langer, supra; Sefton, CRC Crit. Ref Biomed. Eng. 14:20, 1987; Buchwald et al., Surgery 88:507, 1980; Saudek et al., N. Engl. J. Med. 321:574, 1989). Polymeric materials can be used to effect controlled or sustained release therapies of the present invention (see, for example, Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla., 1974; Controlled Drug Bioavailability , Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York, 1984; Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61, 1983; see also Levy et al . ., Science 228:190, 1985; During et al., Ann. Neurol. 25:351, 1989; Howard et al., J. Neurosurg. 7 1:105, 1989; US Patent No. 5,679,377; US Patent No. 5,916,597; US Patent No. 5,912,015 US Patent No. 5,989,463 US Patent No. 5,128,326 PCT Publication No. WO 99/15154 and PCT Publication No. WO 99/20253 Examples of polymers used in sustained release formulations include, without limitation, poly(2- hydroxyethyl methacrylate), poly(methyl methacrylate), poly(acrylic acid), copolymer(ethylene-vinyl acetate), poly(methacrylic acid), polyg lycolides (PLG), polyanhydrides, poly(N-vinylpyrrolidone), poly(vinyl alcohol), polyacrylamide, poly(ethylene glycol), polylactides (PLA), copolymers (lactide and glycolides) (PLGA) and polyorthoesters. In one embodiment, the polymer used in the sustained release formulation is inert, free of leachable impurities, storage stable, sterile, and biodegradable. The controlled or sustained release system can be placed near the prophylactic or therapeutic target, requiring only a fraction of the systemic dose (see, for example, Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138, 1984).

[00342] Controlled release systems are reviewed by Langer, Science 249:1527-1533, 1990). Any technique known to one of skill in the art can be used to prepare sustained release formulations containing one or more of the immunoconjugates of the present invention. See, for example, US Pat. No. 4,526,938, PCT Publication WO 91/05548, PCT Publication WO 96/20698, Ning et al., Radiotherapy & Oncology 39:179-189, 1996; Song et al., PDA Journal of Pharmaceutical Science & Technology 50:372-397, 1995; Cleek et al., Pro. Int'l. Symp. control. Rel. Bioact. mater. 24:853-854, 1997; and Lam et al., Proc. Int'l. Symp. Control Rel. Bioact. mater. 24:759-760, 1997, each of which is incorporated herein by reference in its entirety.

[00343] When the immunoconjugates of the present invention are administered topically, they may be formulated as an ointment, cream, transdermal patch, lotion, gel, spray, aerosol, solution, emulsion, or other form widely known to one of skill in the art. See, for example, Remington's Pharmaceutical Sciences and Introduction to Pharmaceutical Dosage Forms, 19th ed., Mack Pub. Co., Easton, Pa. (1995). In the case of non-sprayable topical dosage forms, viscous-semi-solid or solid forms are usually used, containing a carrier or one or more excipients, compatible with topical application and, in some cases, characterized by a dynamic viscosity greater than that of water. Suitable formulations include, without limitation, solutions, suspensions, emulsions, creams, ointments, powders, liniments, balms, and the like, which are sterilized as necessary or mixed with adjuvants (e.g., preservatives, stabilizers, wetting agents, buffers, or salts) to influence on various properties, such as, for example, osmotic pressure. Other suitable topical dosage forms include nebulized aerosol formulations wherein the active ingredient, in some cases in combination with a solid or liquid inert carrier, is packaged in admixture with a pressurized volatile substance (e.g. a gaseous propellant such as Freon) or in a spray bottle. Humectants or humectants may also be added to pharmaceutical compositions and dosage forms, if desired. Examples of such additional ingredients are well known in the art.

[00344] When compositions containing immunoconjugates are administered intranasally, they may be formulated as an aerosol, spray, mist, or droplet. In particular, for convenience, prophylactic or therapeutic agents for use according to the present invention can be delivered in the form of an aerosol spray from pressurized packets or by means of a nebulizer using a suitable propellant (for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas) . In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges (eg consisting of gelatin) for use in an inhaler or insufflator may be formulated containing a powder mixture of the compound and a suitable powder base such as lactose or starch.

[00345] Methods for co-administration or treatment using a second therapeutic agent, such as a cytokine, steroid, chemotherapeutic agent, antibiotic, or radiation, are known in the art (see, for example, Hardman et al., (eds.) (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, ^10th ed., McGraw-Hill, New York, NY Poole and Peterson (eds.) (2001) Pharmacotherapeutics for Advanced Practice: A Practical Approach, Lippincott, Williams & Wilkins, Phila. , Pa.; Chabner and Longo (eds.) (2001) Cancer Chemotherapy and Biotherapy, Lippincott, Williams & Wilkins, Phila., Pa.). An effective amount of a therapeutic agent can reduce symptoms by at least 10%; at least 20%; at least about 30%; at least 40% or at least 50%.

[00346] Additional therapies (e.g., prophylactic or therapeutic agents) that can be used in combination with the immunoconjugates of the present invention can be used at intervals of less than 5 minutes, at intervals of less than 30 minutes, at intervals of 1 hour, at intervals of approximately 1 hour , at an interval of about 1 to about 2 hours, at an interval of about 2 hours to about 3 hours, at an interval of about 3 hours to about 4 hours, at an interval of about 4 hours to about 5 hours, at an interval of about 5 hours to about 6 hours, from about 6 hours to about 7 hours, from about 7 hours to about 8 hours, from about 8 hours to about 9 hours, from about 9 hours to about 10 hours, from interval from about 10 hours to about 11 hours, with an interval from about 11 hours hours to approximately 12 hours, at intervals of approximately 12 hours to 18 hours, at intervals of 18 hours to 24 hours, at intervals of 24 hours to 36 hours, at intervals of 36 hours to 48 hours, at intervals of 48 hours to 52 hours, at intervals of 52 hours to 60 hours, at intervals of 60 hours to 72 hours, at intervals of 72 hours to 84 hours, at intervals of 84 hours to 96 hours, or at intervals of 96 hours to 120 hours from the administration of immunoconjugates according to the present invention. Two or more therapies may be used during the same patient visit.

[00347] In some embodiments, the implementation of the immunoconjugates of the present invention can be formulated to ensure accurate distribution in vivo . For example, the blood-brain barrier (BBB) keeps many highly hydrophilic compounds out. To ensure that the therapeutic compounds of the present invention cross the BBB (if necessary), they can be formulated, for example, in liposomes. For methods of making a liposome, see, for example, US Pat. Nos. 4,522,811; 5374548 and 5399331. Liposomes may contain one or more fragments that are selectively transported to specific cells or organs, thereby improving targeted drug delivery (see, for example, Ranade, (1989) J. Clin. Pharmacol. 29:685) . Illustrative targeting fragments include folate or biotin (see, for example, US patent No. 5416016 from Low et al.); mannosides (Umezawa et al., (1988) Biochem. Biophys. Res. Commun. 153:1038); antibodies (Bloeman et al., (1995) FEBS Lett. 357:140; Owais et al., (1995) Antimicrob. Agents Chemother. 39:180); surfactant protein A receptor (Briscoe et al., (1995) Am. J. Physiol. 1233:134); p 120 (Schreier et al., (1994) J. Biol. Chem. 269:9090); see also K. Keinanen; ML Laukkanen (1994) FEBS Lett. 346:123; JJ Killion; IJ Fidler (1994) Immunomethods 4:273.

[00348] The present invention provides protocols for administering to a subject in need thereof a pharmaceutical composition containing the immunoconjugates of the present invention, alone or in combination with other therapies. Therapies (eg, prophylactic or therapeutic agents) of the combination therapies of the present invention may be applied to a subject simultaneously or sequentially. Therapies (eg, prophylactic or therapeutic agents) of the combination therapies of the present invention can also be administered cyclically. Cyclic therapy involves administering a first therapy (e.g., a first prophylactic or therapeutic agent) over a period of time and then administering a second therapy (e.g., a second prophylactic or therapeutic agent) over a period of time and repeating such sequential administration, i.e. e. cycle, in order to reduce the development of resistance to one of the therapies (for example, agents), to avoid or reduce the side effects of one of the therapies (for example, agents) and / or to improve the effectiveness of the therapies.

[00349] Therapies (eg, prophylactic or therapeutic agents) of the combination therapies of the present invention may be applied to a subject simultaneously.

[00350] The term "simultaneously" is not limited to the use of therapies (e.g., prophylactic or therapeutic agents) at exactly the same time, but rather means that the pharmaceutical composition containing the antibodies or fragments thereof of the present invention is administered to the subject sequentially and in a period of time such that the antibody-drug conjugates of the present invention can act in conjunction with the other therapy(s) to provide greater benefit than if administered otherwise. For example, each therapy may be applied to a subject at the same time or sequentially in any order at different points in time; however, if they are not used at the same time, they should be used close enough in time to provide the desired therapeutic or prophylactic effect. Each therapy can be administered to a subject individually, in any suitable form, and by any suitable route. In various embodiments, therapies (e.g., prophylactic or therapeutic agents) are administered to the subject at intervals of less than 5 minutes, at intervals of less than 15 minutes, at intervals of less than 30 minutes, at intervals of less than 1 hour, at intervals of approximately 1 hour, at intervals of from about 1 hour to about 2 hours, from about 2 hours to about 3 hours, from about 3 hours to about 4 hours, from about 4 hours to about 5 hours, from about 5 hours to about 6 hours, from about 6 hours to about 7 hours, from about 7 hours to about 8 hours, from about 8 hours to about 9 hours, from about 9 hours to about 10 hours, from approximately 10 hours to approximately 11 hours, at intervals of approximately 11 hours to approximately 12 hour intervals, 24 hour intervals, 48 hour intervals, 72 hour intervals, or 1 week intervals. In other embodiments, two or more therapies (eg, prophylactic or therapeutic agents) are administered during the same patient visit.

[00351] Prophylactic or therapeutic agents of combination therapies may be administered to a subject in the same pharmaceutical composition. Alternatively, the prophylactic or therapeutic agents of the combination therapies may be administered to the subject simultaneously in separate pharmaceutical compositions. Prophylactic or therapeutic agents may be administered to a subject via the same or different routes of administration. The prophylactic or therapeutic agents of the combination therapies may be administered to a subject in the same pharmaceutical composition. Alternatively, the prophylactic or therapeutic agents of the combination therapies may be administered to the subject simultaneously in separate pharmaceutical compositions. Prophylactic or therapeutic agents may be administered to a subject via the same or different routes of administration.

Examples

Example 1 Obtaining antibodies to CCR7

Obtaining Constructs for the Expression of Human, Rat, Mouse, and Cynomolgus Macaque CCR7

[00352] Full-length human, cynomolgus, and mouse CCR7 genes were synthesized based on amino acid sequences from GenBank or Uniprot databases (SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101). Rat CCR7 cDNA template was obtained by gene synthesis based on amino acid sequence information obtained using mRNA isolated from various rat tissues (SEQ ID NO: 103). All synthesized DNA fragments were cloned into appropriate expression vectors.

Table 2. Information about the amino acid and nucleotide sequence of CCR7

human CCR7 SEQ ID NO: 97 mdlgkpmksvlvvallvifqvclcqdevtddyigdnttvdytlfeslcskkdvrnfkawflpimysiicfvgllgnglvvltyiyfkrlktmtdtyllnlavadilflltlpfwaysaakswvfgvhfcklifaiykmsffsgmllllcisidryvaivqavsahrhrarvllisklscvgiwilatvlsipellysdlqrssseqamrcslitehveafitiqvaqmvigflvpllamsfcylviirtllqarnfernkaikviiavvvvfivfqlpyngvvlaqtvanfnitsstcelskqlniaydvtyslacvrccvnpflyafigvkfrndlfklfkdlgclsqeqlrqwsscrhirrssmsveaettttfsp SEQ ID NO: 98 ATGGACCTGGGGAAACCAATGAAAAGCGTGCTGGTGGTGGCTCTCCTTGTCATTTTCCAGGTATGCCTGTGTCAAGATGAGGTCACGGACGATTACATCGGAGACAACACCACAGTGGACTACACTTTGTTCGAGTCTTTGTGCTCCAAGAAGGACGTGCGGAACTTTAAAGCCTGGTTCCTCCCTATCATGTACTCCATCATTTGTTTCGTGGGCCTACTGGGCAATGGGCTGGTCGTGTTGACCTATATCTATTTCAAGAGGCTCAAGACCATGACCGATACCTACCTGCTCAACCTGGCGGTGGCAGACATCCTCTTCCTCCTGACCCTTCCCTTCTGGGCCTACAGCGCGGCCAAGTCCTGGGTCTTCGGTGTCCACTTTTGCAAGCTCATCTTTGCCATCTACAAGATGAGCTTCTTCAGTGGCATGCTCCTACTTCTTTGCATCAGCATTGACCGCTACGTGGCCATCGTCCAGGCTGTCTCAGCTCACCGCCACCGTGCCCGCGTCCTTCTCATCAGCAAGCTGTCCTGTGTGGGCATCTGGATACTAGCCACAGTGCTCTCCATCCCAGAGCTCCTGTACAGTGACCTCCAGAGGAGCAGCAGTGAGCAAGCGATGCGATGCTCTCTCATCACAGAGCATGTGGAGGCCTTTATCACCATCCAGGTGGCCCAGATGGTGATCGGCTTTCTGGTCCCCCTGCTGGCCATGAGCTTCTGTTACCTTGTCATCATCCGCACCCTGCTCCAGGCACGCAACTTTGAGCGCAACAAGGCCATCAAGGTGATCATCGCTGTGGTCGTGGTCTTCATAGTCTTCCAGCTGCCCTACAATGGGGTGGTCCTGGCCCAGACGGTGGCCAACTTCAACATCACCAGTAGCACCTGTGAGCTCAGTAAGCAACTCAACATCGCCTACGACGTCACCTACAGCCTGGCCTGCGTCCGCTGCTGCGTCAACCCTTTCTTGTACGCCTTCATCGGCGTCAAGTTCC GCAACGATCTCTTCAAGCTCTTCAAGGACCTGGGCTGCCTCAGCCAGGAGCAGCTCCGGCAGTGGTCTTCCTGTCGGCACATCCGGCGCTCCTCCATGAGTGTGGAGGCGAGACCACCACCACCTTCTCCCCA CCR7 cynomolgus monkey SEQ ID NO: 99 MDLGKPMKSVLVVALLVIFQVCLCQDEVTDDYIGDNTTVDYTLFESLCSKKDVRNFKAWFLPIMYSIICFVGLLGNGLVVLTYIYFKRLKTMTDTYLLNLAVADILFLLTLPFWAYSAAKSWVFGVHFCKLIFAIYKMSFFSGMLLLLCISIDRYVAIVQAVSAHRHRARVLLISKLSCVGIWILATVLSIPELLYSGLQRSSSEQAMRCSLITEHVEAFITIQVAQMVIGFLVPLLAMSFCYLVIIRTLLQARNFERNKAIKVIIAVVVVFIVFQLPYNGVVLAQTVANFNITSSTCELSKQLNIAYDVTYSLACVRCCVNPFLYAFIGVKFRNDLFKLFKDLGCLSQEQLRQWSSCRHIRRSSMSVEAETTTTFSP SEQ ID NO: 100 ATGGACCTGGGGAAACCAATGAAAAGCGTGCTGGTGGTGGCTCTCCTTGTCATTTTCCAGGTATGCCTGTGTCAAGATGAGGTCACGGACGATTACATCGGAGACAACACCACAGTGGACTACACTTTGTTCGAGTCTTTGTGCTCCAAGAAGGACGTGCGGAACTTTAAAGCCTGGTTCCTCCCTATCATGTACTCCATCATTTGTTTCGTGGGCCTACTGGGCAATGGGCTGGTCGTGTTGACCTATATCTATTTCAAGAGGCTCAAGACCATGACCGATACCTACCTGCTCAACCTGGCGGTGGCAGACATCCTCTTCCTCCTGACCCTTCCCTTCTGGGCCTACAGCGCGGCCAAGTCCTGGGTCTTCGGTGTCCACTTTTGCAAGCTCATCTTTGCCATCTACAAGATGAGCTTCTTCAGTGGCATGCTCCTACTTCTTTGCATCAGCATTGACCGCTACGTGGCCATCGTCCAGGCTGTCTCAGCTCACCGCCACCGTGCCCGCGTCCTTCTCATCAGCAAGCTGTCCTGTGTGGGCATCTGGATACTAGCCACAGTGCTCTCCATCCCAGAGCTCCTGTACAGTGGCCTCCAGAGGAGCAGCAGTGAGCAAGCGATGCGATGCTCTCTCATCACAGAGCATGTGGAGGCCTTTATCACCATCCAGGTGGCCCAGATGGTGATCGGCTTTCTGGTCCCCCTGCTGGCCATGAGCTTCTGTTACCTTGTCATCATCCGCACCCTGCTCCAGGCACGCAACTTTGAGCGCAACAAGGCCATCAAGGTGATCATCGCTGTGGTCGTGGTCTTCATAGTCTTCCAGCTGCCCTACAATGGGGTGGTCCTGGCCCAGACGGTGGCCAACTTCAACATCACCAGTAGCACCTGTGAGCTCAGTAAGCAACTCAACATCGCCTACGACGTCACCTACAGCCTGGCCTGCGTCCGCTGCTGCGTCAACCCTTTCTTGTACGCCTTCATCGGCGTCAAGTTCC GCAACGATCTCTTCAAGCTCTTCAAGGACCTGGGCTGCCTCAGCCAGGAGCAGCTCCGGCAGTGGTCTTCCTGTCGGCACATCCGGCGCTCCTCCATGAGTGTGGAGGCGAGACCACCACCACCTTCTCCCCA CCR7 mice SEQ ID NO: 101 MDPGKPRKNVLVVALLVIFQVCFCQDEVTDDYIGENTTVDYTLYESVCFKKDVRNFKAWFLPLMYSVICFVGLLGNGLVILTYIYFKRLKTMTDTYLLNLAVADILFLLILPFWAYSEAKSWIFGVYLCKGIFGIYKLSFFSGMLLLLCISIDRYVAIVQAVSAHRHRARVLLISKLSCVGIWMLALFLSIPELLYSGLQKNSGEDTLRCSLVSAQVEALITIQVAQMVFGFLVPMLAMSFCYLIIIRTLLQARNFERNKAIKVIIAVVVVFIVFQLPYNGVVLAQTVANFNITNSSCETSKQLNIAYDVTYSLASVRCCVNPFLYAFIGVKFRSDLFKLFKDLGCLSQERLRHWSSCRHVRNASVSMEAETTTTFSP SEQ ID NO: 102 ATGGACCCAGGGAAACCCAGGAAAAACGTGCTGGTGGTGGCTCTCCTTGTCATTTTCCAGGTGTGCTTCTGCCAAGATGAGGTCACCGATGACTACATCGGCGAGAATACCACGGTGGACTACACCCTGTACGAGTCGGTGTGCTTCAAGAAGGATGTGCGGAACTTTAAGGCCTGGTTCCTGCCTCTCATGTATTCTGTCATCTGCTTCGTGGGCCTGCTCGGCAACGGGCTGGTGATACTGACGTACATCTATTTCAAGAGGCTCAAGACCATGACGGATACCTACCTGCTCAACCTGGCCGTGGCAGACATCCTTTTCCTCCTGATTCTTCCCTTCTGGGCCTACAGCGAAGCCAAGTCCTGGATCTTTGGCGTCTACCTGTGTAAGGGCATCTTTGGCATCTATAAGTTAAGCTTCTTCAGCGGGATGCTGCTGCTCCTATGCATCAGCATTGACCGCTACGTAGCCATCGTCCAGGCCGTGTCGGCTCATCGCCACCGCGCCCGCGTGCTTCTCATCAGCAAGCTGTCCTGTGTGGGCATCTGGATGCTGGCCCTCTTCCTCTCCATCCCGGAGCTGCTCTACAGCGGCCTCCAGAAGAACAGCGGCGAGGACACGCTGAGATGCTCACTGGTCAGTGCCCAAGTGGAGGCCTTGATCACCATCCAAGTGGCCCAGATGGTTTTTGGGTTCCTAGTGCCTATGCTGGCTATGAGTTTCTGCTACCTCATTATCATCCGTACCTTGCTCCAGGCACGCAACTTTGAGCGGAACAAGGCCATCAAGGTGATCATTGCCGTGGTGGTAGTCTTCATAGTCTTCCAGCTGCCCTACAATGGGGTGGTCCTGGCTCAGACGGTGGCCAACTTCAACATCACCAATAGCAGCTGCGAAACCAGCAAGCAGCTCAACATTGCCTATGACGTCACCTACAGCCTGGCCTCCGTCCGCTGCTGCGTCAACCCTTTCTTGTATGCCTTCATCGGCGTCAAGTTCC GCAGCGACCTCTTCAAGCTCTTCAAGGACTTGGGCTGTCTCAGCCAGGAACGGCTCCGGCACTGGTCTTCCTGCCGGCATGTACGGAACGCGTCGTGAGCATGGAGGCGGAGACCACCACAACCTTCTCCCCG CCR7 rats SEQ ID NO: 103 MDLGKPTKNVLVVALLVIFQVCFCQDEVTDDYIGENTTVDYTLYESVCFKKDVRNFKAWFLPLMYSVICFVGLLGNGLVVLTYIYFKRLKTMTDTYLLNLAVADILFLMILPFWAYSEAKSWIFGAYLCKSIFGIYKLSFFSGMLLLLCISIDRYVAIVQAVSAHRHRARVLLISKLSCIGIWTLAFFLSIPELLYSGLQKNSGEDTWRCSLVSAQVEALIAIQVAQMVVGFVLPMLAMSFCYLVIIRTLLQARNFERNKAIKVIIAVVVVFVVFQLPYNGVVLAQTVANFNITNSSCEASKQLNIAYDVTYSLASVRCCVNPFLYAFIGVKFRSDLFKLFKDLGCLSQERLRQWSSCRHVRHTSVSMEAETTTTFSP SEQ ID NO: 104 ATGGACCTGGGGAAGCCCACGAAAAACGTGCTGGTGGTGGCTCTCCTGGTCATTTTCCAGGTGTGCTTCTGCCAAGATGAGGTCACAGACGACTACATCGGCGAGAACACCACCGTGGACTACACCCTGTATGAGTCGGTGTGCTTCAAGAAGGATGTGCGGAACTTTAAGGCCTGGTTCCTCCCTCTCATGTACTCAGTCATTTGCTTCGTGGGCCTGCTAGGCAATGGGCTGGTGGTGCTGACATACATCTATTTCAAGAGACTGAAGACCATGACGGATACCTACCTGCTCAACCTGGCCGTGGCAGACATCCTCTTCCTCATGATCCTTCCCTTCTGGGCCTACAGCGAAGCCAAGTCCTGGATCTTTGGTGCCTACCTGTGTAAGAGCATCTTTGGCATCTACAAGTTAAGCTTCTTCAGCGGGATGTTGCTGCTCCTGTGTATCAGCATTGACCGCTATGTGGCCATCGTCCAGGCCGTGTCAGCCCACCGGCACCGCGCCCGCGTGCTTCTCATCAGCAAGCTGTCCTGTATAGGCATCTGGACGCTGGCCTTTTTCCTTTCTATCCCTGAGCTGCTCTACAGCGGCCTCCAGAAGAACAGCGGCGAGGACACGTGGAGATGCTCCCTGGTCAGTGCCCAAGTGGAGGCCTTGATCGCCATCCAAGTGGCCCAGATGGTTGTTGGGTTTGTACTGCCTATGCTGGCTATGAGTTTCTGCTACCTGGTTATCATCCGCACTCTGCTCCAGGCGCGAAACTTCGAGCGGAACAAGGCCATCAAGGTGATCATCGCTGTGGTCGTAGTGTTCGTCGTCTTCCAGCTGCCCTACAATGGGGTGGTCCTGGCCCAGACCGTGGCCAATTTCAACATCACCAATAGCAGCTGCGAAGCCAGCAAGCAGCTCAACATTGCCTATGACGTCACCTACAGCCTGGCCTCCGTCCGCTGCTGTGTCAACCCTTTCTTGTATGCCTTCATCGGCGTCAAGTTCC GCAGCGACCTCTTCAAGCTCTTCAAGGACTTGGGCTGCCTCAGCCAGGAACGGCTCCGGCAGTGGTCTTCCTGCCGCCATGTACGGCACACGTCCGTGAGCATGGAGGCGGAGACTACCACCACCTTCTCCCCG

Generation of cell lines stably expressing CCR7

[00353] Cell lines stably expressing CCR7 were obtained using retroviral transduction. 293T cells were co-transfected with the CCR7 retroviral expression vector and packaging vector pCL-Eco or pCL-10A1 (Novus, USA, cat. no. NBP2-29540 or NBP2-2952) using Fugene 6 transfection reagent (Promega, USA, cat. no. . E2692) following the manufacturer's recommendations. Cells were incubated at 37° C. in a humidified CO ₂ incubator and viral supernatant was collected 48 hours after transfection. NIH/3T3 and 300.19 cells were grown to a nearly confluent monolayer. Growth medium was removed from cells and viral supernatant was added in the presence of 8 μg polybrene/ml (final concentration) (EMD Millipore, Cat # TR-1003-G). After incubation for 3-6 hours at 37°C, fresh medium was added. The cells were then cultured under conditions suitable for selection to obtain cell lines stably expressing CCR7.

Production, expression and purification of virus-like particles (VLP)

[00354] HEK293T or NIH/3T3 cells were maintained in DMEM with 10% FBS. To obtain VLPs, cells were medium changed to DMEM with 4% FBS, then co-transfected with CCR7 expression plasmid and Gag retroviral expression plasmid at a ratio of 3:2 in µg. Forty-eight hours after transfection, cell supernatant was collected and clarified by centrifugation at 2500× g for 5 min in a bench top centrifuge and kept on ice. VLPs were purified by ultracentrifugation at 100,000× g through a 20% sucrose pad in 38 ml Beckman Ultra-Clear centrifuge tubes (p/n 344058) on a Beckman Coulter SW 32 Ti-rotor on a Sorvall RC 6+ ultracentrifuge. The pellets obtained were resuspended in 300 μl of cold sterile PBS and quantified using the BCA assay (Pierce, cat. no. 23225).

Creation of the CCR7 immunogenic backbone based on the structure

[00355] Members of the G-protein coupled receptor family are membrane proteins that contain seven transmembrane helical regions (TM1...TM7), each connected by binding sequences of varying length. The amino-terminus of the protein is located on the outside of the cell surface, which means that 4 regions of the protein are potentially located on the cell surface, the amino-terminus (N-terminus) and 3 extracellular loop regions (EC1, EC2 and EC3). Thus, such regions are available as antigens for antibodies.

[00356] It was contemplated that a combination of one or more of these 4 elements could be inserted into the soluble protein backbone to structurally approximate the CCR7 region outside the cell.

[00357] To determine the optimal extracellular regions of CCR7, a model was built using the crystal structure of a close homologue of CXCR4 (Wu et al., "Structures of the CXCR4 chemokine GPCR with small-molecule and cyclic peptide antagonists". (2010) Science 330: 1066-1071 ) by combining the CXCR4 structure with protein structure database entries (3ODU, 3OE0, 3OE6, 3OE8, 3OE9) and modeling software. Based on the model, it was concluded that the amino acids that are located between the binding transmembrane helical regions are located on the surface of the protein. These areas are identified in Table 3 below.

Table 3. Information on the amino acid and nucleotide sequence of the CCR7 immunogenic backbone

SEQID NO: Description Comments 105 CCR7
(NP_001829.1| chemokine receptor type 7 with CC motif) MDLGKPMKSVLVVALLVIFQV CLC QDEVTDDYIGDNTTVDYT LFESLCSKKDVR NFKAWFLPIM YSIICFVGLLGNGLVVLTYIYFK RLKTMTDTYLLNLAVADILFLL TLPFWAYSAAKSWVFGVHFCK LIFAIYKMSFFSGMLLLLCISIDR YVAIVQAVSAHRHRARVLLISK LSCVGIWILATVLSIPELL YSDL QRSSSEQAMRCSLIT EHVEAFIT IQVAQMVIGFLVPLLAMSFCYL VIIRTLLQARNFERNKAIKVIIA VVVVFIVFQLPYNGVVLAQTV ANFNITSSTCELSKQLNIAYDVT YSLACVRCCVNPFLYAFIGVKF RNDLFKLFKDLGCLSQEQLRQ WSSCRHIRRSSMSVEAETTTTF SP CCR7, predecessor.
Extracellular regions are highlighted in bold.
Insertion regions or their derivatives are indicated in bold and underlined. 106 N-terminus QDEVTDDYIGDNTTVDYTLFESLCSKKDVR N-terminal extracellular sequence of CCR7 107 EC1 KSWVFGVH Extracellular loop 1 CCR7 108 EC2 YSDLQRSSSEQAMRCSLIT Extracellular loop 2 CCR7 109 EC3 FNITSST Extracellular loop 3 CCR7 110 EC2_C24S YSDLQRSSSEQAMRSSLIT 111 H_MGFTX1 EVQLVESGGGLVKPGGSLKLSCAASGFTFSDYGMLWVRQAPEKGLEWIAYISSGSSTIYYADRVKGRFTISRDNAKNTLFLQMTSLRSEDTAMYYCSTGTFAYWGQGTPVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPPRVTCNVAHPASSTKVDKKK Heavy backbone chain of mouse Fab 112 L_MGFTX1 DVVMTQNPLSLPVSLGDQASISCRSSQSLIYNNGNTYLHWYRQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPFTFGSGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLECTKDEYERHNSYTC Mouse Fab backbone light chain 113 H_FabCCR7M1 QDEVTDDYIGDNTTVDYTLFESLCSKKDVR EVQLVESGGGLVKPGGSLKLSCAASGFTFSDYGMLWVRQAPEKGLEWIAYISSGSSTIYYADRVKGRFTISRDNAKNTLFLQMTSLRSEDTAMYYCSTGT YSDLQRSSSEQAMRSSLIT FAYWGQGTPVTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDC Heavy chain Fab with inserted N-terminus and EC2
Inserted sequences are underlined and indicated in bold 114 H_FabCCR7M1 CAAGATGAGGTCACGGACGATTACATCGGAGACAACACCACAGTGGACTACACTTTGTTCGAGTCTTTGTGCTCCAAGAAGGACGTGCGGgaggtgcagctggtggagtctggtggtggtctggtcaagcctggaggttccctgaaactgagttgtgccgcatctgggtttacattctctgactacggaatgctgtgggtgaggcaggcaccagagaagggcctggaatggatcgcttatatttccagcggatctagtactatctactatgcagacagggtcaagggccggttcaccattagcagagataacgccaaaaataccctgtttctgcagatgacatcactgaggtccgaggataccgctatgtattattgctccacagggactTACAGTGACCTCCAGAGGAGCAGCAGTGAGCAAGCGATGCGATCCTCTCTCATCACAtttgcttactggggacaggggacacccgtgaccgtcagctcagccaagaccaccccccccagcgtgtaccctctggcccctggctctgccgcccagaccaacagcatggtgaccctgggctgcctggtgaagggctacttccccgagcccgtgaccgtgacctggaacagcggcagcctgagcagcggcgtgcacaccttccccgccgtgctgcagagcgacctgtacaccctgagcagctctgtgaccgtgcccagcagcacctggcccagcgagaccgtgacatgcaacgtggcccaccccgccagctccaccaaggtggacaagaaaatcgtgccccgggactgc Heavy chain DNA sequence of HFabCCR7M1. The inserted sequence is indicated by uppercase 115 L_MGFTX1 atgtcgtgatgactcagaatccactgtccctgcctgtgtccctgggcgatcaggcttccattagctgtcgttcctctcagtccctgatctacaacaatggtaacacctacctgcactggtatagacagaagcccggccagtcccctaagctgctgatctacaaagtgagtaataggttctcaggagtcccagaccggttttccggcagcggatctgggaccgatttcacactgaaaatctctagggtggaggccgaagacctgggcgtctacttttgtagtcagagcactcacgtccccttcaccttcggcagcggaacaaaactggaaatcaagcgcgctgatgccgcccctaccgtgagcatcttcccccccagcagcgagcagctgaccagcggcggagccagcgtggtgtgcttcctgaacaacttctaccccaaggacatcaacgtgaagtggaagatcgacggcagcgagcggcagaacggcgtgctgaacagctggaccgaccaggacagcaaggactccacctacagcatgagcagcaccctgaccctgaccaaggacgagtacgagcggcacaacagctacacctgcgaggccacccacaagaccagcaccagccccatcgtgaagagcttcaaccggaacgagtgc DNA sequence L_MGFTX1

[00358] The small size of the EC1 and EC3 loops and the spatial separation of EC3 from the other three regions prioritizes the use of the N-terminus and the EC2 loop as candidate epitopes.

[00359] Simulation of the fusion of the N-terminus sequence to the mouse Fab crystal structure with the EC2 sequence inserted into different regions of the Fab loops, such as framework 1, CDR-H3 or CDR3-H1, showed that if the degree of flexibility for data two sequences is valid, then they may represent reasonable approximations for the region data structure in CCR7.

Creation of an immunogenic scaffold for immunization of mice

[00360] Graft constructs for immunization of mice were generated by fusing the N-terminal sequence from Table 3 to the N-terminus of the heavy chain of the mouse Fab backbone (designated MGFTX1) and grafting a modified EC2 sequence variant with a cysteine residue mutation at position 24 to serine (Table 3 and underlined+bold sequences in SEQ ID NO: 113) in CDR3 from the backbone of MGFTX1, then both heavy and light chains were produced from the immunoglobulin chains to generate final protein constructs. Thus, the constructs, designated FabCCR7M1, contained a murine framework region that should be immune tolerant when combined with a human sequence to which the mouse immune response was directed.

[00361] The N-terminal sequence was directly fused to the heavy chain of the MGFTX1 backbone. Insertion points for EC2 were chosen to be in the middle of the CDR loop based on the available data for the structural or homologous model. Fab-grafted proteins were generated using standard molecular biology techniques using recombinant DNA encoding the appropriate sequences.

[00362] For example, the variable region of each antibody containing EC2 inserted into the heavy chain CDR3 was synthesized. The DNA encoding the variable region was amplified by PCR, and the resulting fragment was subcloned into a vector containing either the light chain constant region or the heavy chain constant region and Fc region. In this manner, FabCCR7M1 proteins were obtained with the appropriate N-terminus fusion and EC2 insertion into H3. The resulting constructs are shown in Table 3. Transfections using suitable combinations of heavy and light chain vectors result in the expression of a recombinant Fab containing one N-terminal molecule and one EC2 molecule.

[00363] The choice of which CDR is selected for grafting is determined based on the parameters of the external location of the loop and proximity to the N-terminus. At present, simulation software is only partially applicable to predict which CDR and which location within the CDR will provide the required parameters due to the flexibility of loops after merging and grafting. The structure of the backbone of MGFTX1 in combination with EC2_C24S (Table 3, SEQ ID NO: 110) showed that the sequence is external and flexible, as evidenced by the lack of electron density in most of the sequence.

[00364] To summarize, the insertion point in the CDR was chosen based on structure, with the assumption that grafting into the CDR would provide some level of structural similarity to the native antigen.

Creating a hybridoma

[00365] Bcl-2 transgenic mice (line C57BL/6-Tgn (bcl-2) 22 WEHI) were immunized with antigen using a procedure called Multiple Site Boosting Immunization (RIMMS) (McIntyre GD. Hybridoma 1997). Briefly, mice were injected with 1-3 μg of CCR7 immunogen at 8 specific sites close to the peripheral lymph nodes (PLN). This procedure was repeated 8 times over a 12 day period. On day 12, blood samples were collected and serum antibody titer was analyzed by FACS. In some cases, BALB/c and/or C57Bl/6 mice were immunized with NIH3T3 or 300.19 cells stably overexpressing human CCR7 (SEQ ID NO: 97). Animals were injected subcutaneously with 5×10 ⁶ cells in PBS once a month for 3 months, followed by an intravenous boost injection of 25 μg VLP expressing human CCR7. Two days after the booster injection, blood samples were collected and serum antibody titer was analyzed by FACS. Spleens and pooled PLNs were recovered from mice with high titers. To collect lymphocytes, spleen and PLN were washed twice with DMEM and then separated into cells by passing through a 70 micron sieve (Falcon, no. 352350). The resulting lymphocytes were washed 2 more times before confluence in Cytofusion medium (BTXpress Cytofusion® electroporation medium, Cat # 47001).

[00366] For fusion, F0 myeloma cells were mixed with lymphocytes at a ratio of 1:4. The cell mixture was centrifuged, suspended in Cytofusion medium and then introduced into an electrofusion chamber (9ML coaxial chamber from Harvard Apparatus, item # 470020). Electrofusion was performed according to the manufacturer's instructions using a Hybrimune hybridoma system model CEEF-50B (Cyto Pulse Sciences, Inc.). The fused cells were left to recover for 5 min in the chamber, diluted 1/10 in HAT-free fusion medium (DMEM+20% FBS, 1% penicillin/streptomycin/glutamine, 1X NEAA, 0.5X HFCS) and placed at 37° C for one hour. 4X HAT medium (DMEM+20% FBS, 1% penicillin/streptomycin/glutamine, 1X NEAA, 4X HAT, 0.5X HFCS) was added to make a 1X solution and the density was adjusted to 1.67×10 ⁴ cells/mL. Cells were seeded in 384-well plates at the rate of 60 μl/well.

Screening with FACS

[00367] Ten days after fusion, hybridoma plates were screened for the presence of CCR7-specific antibodies using flow cytometry to confirm specific binding of candidate antibodies to cell lines stably overexpressing or expressing endogenous CCR7. Cells were thoroughly rinsed with PBS and treated with Accutase (Millipore, #SCR005) to separate from growth plates and resuspended in cold PBS. Cells were labeled with biotin and fluorescent dye according to the manufacturer's instructions (FluoReporter cell surface biotinylation kit, Thermo Fisher Scientific, cat. no. F-20650; streptavidin PE-Cy7, ThermoFisher Scientific, cat. no. SA1012). Cells were resuspended at approximately 1×10 ⁶ cells/ml in FACS buffer (1X DPBS, 3% FBS, 5 mM EDTA, 0.1% sodium azide). In a 384-well plate, 20 µl of the hybridoma supernatant was preliminarily added, and 20 µl of the cell suspension was added. Cells were incubated for 1 hour at 4°C, washed twice with cold FACS buffer and resuspended in 20 μl of secondary antibody diluted 1:400 in FACS buffer (conjugated with goat allophycocyanin F(ab')2 to human IgG, Fcγ- specific; Jackson Immunoresearch, Cat # 109-136-098). After an additional 45 min incubation at 4° C., cells were washed twice with FACS buffer and resuspended in 20 μl of FACS buffer + 2 μg/ml propidium iodide (Sigma Aldrich, Cat # P4864-10ML). The geometric mean of fluorescence intensity was calculated on live single cells using FlowJo™ software.

Purification of antibodies

[00368] Received chimeric antibodies containing mouse variable regions and human constant regions. In addition, chimeric variants containing cysteine mutations (eg, cysteines at position K36°C or positions E152C and S375C of the heavy chain) have been designed for conjugation to a drug moiety to produce the ADCs described in more detail herein. For each of the selected hybridomas, mAb121G12, mAb506E15, mAb674J13 and mAb684E12, the hybridoma-produced antibody variable region (VH and VL) DNA sequences were generated to generate optimized sequences (eg, humanization, preferential characterization). Variable region DNA from mouse monoclonal antibodies was amplified by RACE based on RNA obtained from each selected hybridoma cell line using standard methods. The polypeptide sequences of each of the mouse heavy/light chain variable regions are shown under SEQ ID NO: 128/SEQ ID NO: 144, SEQ ID NO: 160/SEQ ID NO: 176, SEQ ID NO: 192/SEQ ID NO: 208 and SEQ ID NO: 224/SEQ ID NO: 240, respectively, for each of hybridomas 674J13, 121G12, 506E15 and 684E12. The respective derived heavy/light chain variable region nucleotide sequences for each of the hybridomas are shown under SEQ ID NO: 129/SEQ ID NO: 145, SEQ ID NO: 161/SEQ ID NO: 177, SEQ ID NO: 193/SEQ ID NO: 209 and SEQ ID NO: 225/SEQ ID NO: 241. To obtain chimeric antibodies, DNA sequences encoding the VL and VH domain of antibodies from hybridoma were subcloned into expression vectors containing the corresponding sequences of the human wild-type heavy chain or the heavy chain with the introduced pathway. constructing a Cys or D265A/P329A (DAPA) mutated heavy chain and human light chain constant region sequence (IgG1, kappa chain).

Humanization of antibodies

[00369] Variable region constructs have been designed to humanize and optimize sequences (e.g., remove post-translational modifications, unwanted sites, etc.), to include cysteine mutations (e.g., cysteines at position K36°C or at positions E152C and S375C of the heavy chain ) for conjugating the drug moiety and producing the ADCs described in more detail herein; and to modify with Fc effector region mutations (eg, D265A/P329A mutations in the Fc region) to provide constructs having reduced Fc effector function and to provide combinations thereof.

[00370] DNA sequences encoding humanized VL and VH domains were ordered from GeneArt (Life Technologies Inc., Regensburg, Germany), including codon optimization for Cricetulus griseus. The sequences encoding the VL and VH domains were subcloned from vectors taken from GeneArt into expression vectors suitable for protein production in mammalian cells. The heavy and light chains were cloned into separate expression vectors to allow co-transfection.

[00371] Recombinant antibodies (IgG1, kappa chain) were generated by co-transfection of vectors into Freestyle™ 293 expressing cells (Invitrogen, USA) using PEI (polyethyleneimine, MW 25000, linear, Polysciences, USA, Cat # 23966) as a transfection reagent. A stock solution of PEI was prepared by dissolving 1 g of PEI in 900 ml of water suitable for cell culture at room temperature (RT). To facilitate dissolution of the PEI, the solution was acidified with HCl to pH 3-5 followed by neutralization with NaOH to give a final pH of 7.05. Finally, the volume was adjusted to 1 L and the solution was sterilized by filtration through a 0.22 μm filter, aliquoted and frozen at -80° C. until further use.

[00372] Freestyle™ 293 cells (Gibco™, ThermoFisher scientific, USA, cat. no. R79007) were cultured in Freestyle™ 293 medium (Gibco™, ThermoFisher scientific, USA, cat. no. 12338018) in shake flasks (Corning, Tewkesbury , Massachusetts) on an orbital shaker (100-120 rpm) at 37°C in a humidified incubator at 5% CO ₂ . In the case of transient transfections, cells were grown to a density of approximately 3×10 ⁶ cells/ml, and then 1 μg filter-sterilized DNA/ml culture (0.5 μg heavy chain + 0.5 μg light chain) was added to a solution of 2 μg PEI/1 μg of DNA in OptiMem (ThermoFisher Scientific, USA, #11058021) and incubated at RT for 8 minutes. The mixture was added dropwise to Freestyle™ 293 cells with gentle mixing in a circular motion. After transfection, the cells were cultured for one to two weeks before purification of the antibody from the supernatant. To establish stable cell lines for antibody production, the vectors were co-transfected by nucleofection (Nucleofector™ 96-well shuttle™; Lonza) into CHO cells using the manufacturer's recommendations and cultured under selective conditions for up to four weeks in shake flasks. Cells were harvested by centrifugation and the supernatant was recovered for antibody purification. The antibody was purified using Protein A, Protein G or MabSelect SuRe columns (GE Healthcare Life Sciences). The resin was equilibrated with PBS before loading the supernatant. After sample binding, the column was washed with PBS and the antibody was eluted with IgG Thermo Elution Buffer (Pierce), pH 2.8 (Cat. No. 21004). The eluted fractions were neutralized with tribasic sodium citrate dehydrate buffer, pH 8.5 (Sigma Aldrich, Cat # S4641-1Kg), and then dialyzed overnight in PBS, pH 7.2.

Brief description of antibodies

[00373] Table 4 sets out the relevant sequence information for the original and humanized anti-CCR7 antibodies derived from mouse hybridoma antibodies. Throughout this application, when describing antibodies, the terms "hybridoma" and "original" are used interchangeably and refer to Abs that are derived from a hybridoma.

Table 4. Amino acid and nucleotide sequence information for hybridoma and humanized anti-CCR7 antibodies

Original 674J13, hIgG1 SEQ ID NO: 116 HCDR1 (combined) GYSITSGYSWH SEQ ID NO: 117 HCDR2 (combined) HIHSSGSTNYNPSLKS SEQ ID NO: 118 HCDR3 (combined) GGVQAFAY SEQ ID NO: 119 HDDR1 (Kabat) SGYSWH SEQ ID NO: 120 HDDR2 (Kabat) HIHSSGSTNYNPSLKS SEQ ID NO: 121 HDDR3 (Kabat) GGVQAFAY SEQ ID NO: 122 HCDR1 (Chothia) GYSITSGY SEQ ID NO: 123 HCDR2 (Chothia) HSSGS SEQ ID NO: 124 HCDR3 (Chothia) GGVQAFAY SEQ ID NO: 125 HCDR1 (IMGT) GYSITSGYS SEQ ID NO: 126 HCDR2 (IMGT) IHSSGST SEQ ID NO: 127 HDDR3 (IMGT) ARGGVQAFAY SEQ ID NO: 128 vh DVQLQESGPDLVKPSQSLSLTCTVTGYSITSGYSWHWIRQFPGNKLEWMAHIHSSGSTNYNPSLKSRISIIRDTSKNLFFLQLNSVTTEDTATYYCARGGVQAFAYWGQGTLVTVSA SEQ ID NO: 129 DNA VH GATGTGCAGCTTCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTCAGTCACTTTCACTCACCTGCACTGTCACTGGCTACTCCATCACCAGTGGTTATAGCTGGCACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGCCCACATCCACTCCAGTGGTAGCACTAACTACAACCCATCTCTCAAAAGTCGCATCTCTATCATTCGAGACACATCCAAGAACCTGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGAGGGGGGGTACAGGCCTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA SEQ ID NO: 130 Heavy chain (Fc WT) DVQLQESGPDLVKPSQSLSLTCTVTGYSITSGYSWHWIRQFPGNKLEWMAHIHSSGSTNYNPSLKSRISIIRDTSKNLFFLQLNSVTTEDTATYYCARGGVQAFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 131 heavy chain DNA gatgtgcagcttcaggagtcaggacctgacctggtgaaaccttctcagtcactttcactcacctgcactgtcactggctactccatcaccagtggttatagctggcactggatccggcagtttccaggaaacaaactggagtggatggcccacatccactccagtggtagcactaactacaacccatctctcaaaagtcgcatctctatcattcgagacacatccaagaacctgttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaagagggggggtacaggcctttgcttactggggccaagggactctggtcactgtctctgcaGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGCCAGCCCCAATCGAAA AGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 132 LCDR1 (combined) SASSSVIYMH SEQ ID NO: 133 LCDR2 (combined) DTSKLAS SEQ ID NO: 134 LCDR3 (combined) QQWSSNPLT SEQ ID NO: 135 LCDR1 (Kabat) SASSSVIYMH SEQ ID NO: 136 LCDR2 (Kabat) DTSKLAS SEQ ID NO: 137 LCDR3 (Kabat) QQWSSNPLT SEQ ID NO: 138 LCDR1 (Chothia) SSSVIY SEQ ID NO: 139 LCDR2 (Chothia) DTS SEQ ID NO: 140 LCDR3 (Chothia) WSSNPL SEQ ID NO: 141 LCDR1 (IMGT) SSVIY SEQ ID NO: 142 LCDR2 (IMGT) DTS SEQ ID NO: 143 LCDR3 (IMGT) QQWSSNPLT SEQ ID NO: 144 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVIYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTTLELK SEQ ID NO: 145 DNA VL CAAATTGTCCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGTTCAAGTGTAATTTACATGCACTGGTACCAGCAGAAGTCAGGCACCTCCCCCAAAAGATGGATTTATGACACATCCAAACTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGTAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCGCTCACGTTCGGTGCTGGGACCACGTTGGAGCTGAAA SEQ ID NO: 146 light chain QIVLTQSPAIMSASPGEKVTMTCSASSSVIYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTTLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACENSFTHQGL SEQ ID NO: 147 light chain DNA CAAATTGTCCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGTTCAAGTGTAATTTACATGCACTGGTACCAGCAGAAGTCAGGCACCTCCCCCAAAAGATGGATTTATGACACATCCAAACTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGTAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCGCTCACGTTCGGTGCTGGGACCACGTTGGAGCTGAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 121G12, hIgG1 SEQ ID NO: 148 HCDR1 (combined) GFTFSTYAMS SEQ ID NO: 149 HCDR2 (combined) TISDGGSYSYYPDNVKG SEQ ID NO: 150 HCDR3 (combined) RGSRYEEYYVMDY SEQ ID NO: 151 HDDR1 (Kabat) TYAMS SEQ ID NO: 152 HDDR2 (Kabat) TISDGGSYSYYPDNVKG SEQ ID NO: 153 HDDR3 (Kabat) RGSRYEEYYVMDY SEQ ID NO: 154 HCDR1 (Chothia) GFTFSTY SEQ ID NO: 155 HCDR2 (Chothia) SDGGSY SEQ ID NO: 156 HCDR3 (Chothia) RGSRYEEYYVMDY SEQ ID NO: 157 HCDR1 (IMGT) GFTFSTYA SEQ ID NO: 158 HDDR2 (IMGT) ISDGGSYS SEQ ID NO: 159 HDDR3 (IMGT) ARRGSRYEEYYVMDY SEQ ID NO: 160 vh EVQLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWVATISDGGSYSYYPDNVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCARRGSRYEEYYVMDYWGQGTSVTVSS SEQ ID NO: 161 DNA VH GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGGTCGCAACCATTAGTGATGGTGGTAGTTATTCGTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTATACCTGCAAATGAGCCATCTGAAGTCTGAGGACACAGCCATGTATTACTGTGCAAGACGAGGTAGTAGGTACGAAGAGTACTATGTTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA SEQ ID NO: 162 heavy chain
(FC WT) EVQLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWVATISDGGSYSYYPDNVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCARRGSRYEEYYVMDYWGQGTSVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 163 heavy chain DNA GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGGTCGCAACCATTAGTGATGGTGGTAGTTATTCGTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTATACCTGCAAATGAGCCATCTGAAGTCTGAGGACACAGCCATGTATTACTGTGCAAGACGAGGTAGTAGGTACGAAGAGTACTATGTTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGC CAGCCCCAATCGAAAAGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 164 LCDR1 (combined) RASQSISNNLH SEQ ID NO: 165 LCDR2 (combined) YASQSIS SEQ ID NO: 166 LCDR3 (combined) QQSNSWLT SEQ ID NO: 167 LCDR1 (Kabat) RASQSISNNLH SEQ ID NO: 168 LCDR2 (Kabat) YASQSIS SEQ ID NO: 169 LCDR3 (Kabat) QQSNSWLT SEQ ID NO: 170 LCDR1 (Chothia) SQSISNN SEQ ID NO: 171 LCDR2 (Chothia) YAS SEQ ID NO: 172 LCDR3 (Chothia) SNSWL SEQ ID NO: 173 LCDR1 (IMGT) QSISNN SEQ ID NO: 174 LCDR2 (IMGT) YAS SEQ ID NO: 175 LCDR3 (IMGT) QQSNSWLT SEQ ID NO: 176 VL DIVLTQSPATLSVTPGDSVSLSCRASQSISNNLHWYQQKSHESPKLLIKYASQSISGIPSRFSGSGSGTDFTLSINSVETEDFGMYFCQQSNSWLTFGAGTKLGLK SEQ ID NO: 177 DNA VL GATATTGTGCTAACTCAGTCTCCAGCCACCCTGTCTGTGACTCCAGGAGATAGCGTCAGTCTTTCCTGCAGGGCCAGCCAAAGTATTAGCAACAACCTACACTGGTATCAACAGAAATCACATGAGTCTCCAAAACTTCTCATCAAGTATGCTTCCCAGTCCATCTCTGGGATCCCCTCCAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACTCTCAGTATCAACAGTGTGGAGACTGAAGATTTTGGAATGTATTTCTGTCAACAGAGTAACAGCTGGCTCACGTTCGGTGCTGGGACCAAGCTGGGGCTGAAA SEQ ID NO: 178 light chain DIVLTQSPATLSVTPGDSVSLSCRASQSISNNLHWYQQKSHESPKLLIKYASQSISGIPSRFSGSGSGTDFTLSINSVETEDFGMYFCQQSNSWLTFGAGTKLGLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACESFTHQGLSSPV SEQ ID NO: 179 light chain DNA GATATTGTGCTAACTCAGTCTCCAGCCACCCTGTCTGTGACTCCAGGAGATAGCGTCAGTCTTTCCTGCAGGGCCAGCCAAAGTATTAGCAACAACCTACACTGGTATCAACAGAAATCACATGAGTCTCCAAAACTTCTCATCAAGTATGCTTCCCAGTCCATCTCTGGGATCCCCTCCAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACTCTCAGTATCAACAGTGTGGAGACTGAAGATTTTGGAATGTATTTCTGTCAACAGAGTAACAGCTGGCTCACGTTCGGTGCTGGGACCAAGCTGGGGCTGAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 506E15, hIgG1 SEQ ID NO: 180 HCDR1 (combined) GFTFSSYAMS SEQ ID NO: 181 HCDR2 (combined) TISSGGSFTYYPDSVKG SEQ ID NO: 182 HCDR3 (combined) RASTVVGTDFDV SEQ ID NO: 183 HDDR1 (Kabat) SYAMS SEQ ID NO: 184 HDDR2 (Kabat) TISSGGSFTYYPDSVKG SEQ ID NO: 185 HDDR3 (Kabat) RASTVVGTDFDV SEQ ID NO: 186 HCDR1 (Chothia) GFTFSSY SEQ ID NO: 187 HCDR2 (Chothia) SSGGSF SEQ ID NO: 188 HCDR3 (Chothia) RASTVVGTDFDV SEQ ID NO: 189 HCDR1 (IMGT) GFTFSSYA SEQ ID NO: 190 HDDR2 (IMGT) ISSGGSFT SEQ ID NO: 191 HDDR3 (IMGT) ARRASTVVGTDFDV SEQ ID NO: 192 vh EVMLVESGGGLVKPGGSLKLSCAASGFTFSSYAMSWIRQTPEKRLEWVATISSGGSFTYYPDSVKGRFTISRDNVKNTLYLQMSSLRSEDTAMYYCARRASTVVGTDFDVWGAGTTVTVSS SEQ ID NO: 193 DNA VH GAAGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATGCCATGTCTTGGATTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATCAGTAGTGGTGGTAGTTTCACCTACTATCCAGACAGTGTGAAGGGGCGATTCACCATTTCTAGAGACAATGTCAAGAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAAGACACGGCCATGTATTACTGTGCAAGACGGGCTTCTACGGTAGTAGGTACGGACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 194 heavy chain
(FC WT) EVMLVESGGGLVKPGGSLKLSCAASGFTFSSYAMSWIRQTPEKRLEWVATISSGGSFTYYPDSVKGRFTISRDNVKNTLYLQMSSLRSEDTAMYYCARRASTVVGTDFDVWGAGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 195 heavy chain DNA GAAGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATGCCATGTCTTGGATTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATCAGTAGTGGTGGTAGTTTCACCTACTATCCAGACAGTGTGAAGGGGCGATTCACCATTTCTAGAGACAATGTCAAGAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAAGACACGGCCATGTATTACTGTGCAAGACGGGCTTCTACGGTAGTAGGTACGGACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGCCAG CCCCAATCGAAAAGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 196 LCDR1 (combined) RASQDIGSSLN SEQ ID NO: 197 LCDR2 (combined) ATSSLDS SEQ ID NO: 198 LCDR3 (combined) LQYASSPPT SEQ ID NO: 199 LCDR1 (Kabat) RASQDIGSSLN SEQ ID NO: 200 LCDR2 (Kabat) ATSSLDS SEQ ID NO: 201 LCDR3 (Kabat) LQYASSPPT SEQ ID NO: 202 LCDR1 (Chothia) SQDIGSS SEQ ID NO: 203 LCDR2 (Chothia) ATS SEQ ID NO: 204 LCDR3 (Chothia) YASSPP SEQ ID NO: 205 LCDR1 (IMGT) QDIGSS SEQ ID NO: 206 LCDR2 (IMGT) ATS SEQ ID NO: 207 LCDR3 (IMGT) LQYASSPPT SEQ ID NO: 208 VL DIQMTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSGSRSGSDYSLTISSLESEDFVVYYCLQYASSPPTFGGGTKLEIK SEQ ID NO: 209 DNA VL GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTATGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGGCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGTCTATTACTGTCTACAATATGCTAGTTCGCCTCCGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA SEQ ID NO: 210 light chain DIQMTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSGSRSGSDYSLTISSLESEDFVVYYCLQYASSPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKSSVY SEQ ID NO: 211 light chain DNA GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTATGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGGCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGTCTATTACTGTCTACAATATGCTAGTTCGCCTCCGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 684E12, hIgG1 SEQ ID NO: 212 HCDR1 (combined) GFTFSNFAMS SEQ ID NO: 213 HCDR2 (combined) TISTGGTYTYYPDSVKG SEQ ID NO: 214 HCDR3 (combined) RGYDGVDK SEQ ID NO: 215 HDDR1 (Kabat) SNFAMS SEQ ID NO: 216 HDDR2 (Kabat) TISTGGTYTYYPDSVKG SEQ ID NO: 217 HDDR3 (Kabat) RGYDGVDK SEQ ID NO: 218 HCDR1 (Chothia) GFTFSNF SEQ ID NO: 219 HCDR2 (Chothia) STGGTY SEQ ID NO: 220 HCDR3 (Chothia) RGYDGVDK SEQ ID NO: 221 HCDR1 (IMGT) GFTFSNFA SEQ ID NO: 222 HDDR2 (IMGT) ISTGGTYT SEQ ID NO: 223 HDDR3 (IMGT) TRRGYDGVDK SEQ ID NO: 224 vh EVHLVESGGGLVKPGGSLKLSCAASGFTFSNFAMSWVRQTPEKRLEWVATISTGGTYTYYPDSVKGRFTISRDNAKKTLYLQMSSLRSEDTAMYYCTRRGYDGVDKWGQGTTLTVSS SEQ ID NO: 225 DNA VH gaagtgcatctggtggagtctgggggaggcttagtgaagcctggagggtccctgaaactctcctgtgcagcctctggattcactttcagtaactttgccatgtcttgggttcgccagactccggagaagagactggagtgggtcgcaaccattagtactggtggtacttacacctactatccagacagtgtgaagggtcgattcaccatctccagagacaatgccaagaaaaccctgtacctgcaaatgagcagtctgaggtctgaggacacggccatgtattactgtacaagacgggggtacgacggcgtggacaaatggggccaaggcaccactctcacagtctcctca SEQ ID NO: 226 heavy chain
(FC WT) EVHLVESGGGLVKPGGSLKLSCAASGFTFSNFAMSWVRQTPEKRLEWVATISTGGTYTYYPDSVKGRFTISRDNAKKTLYLQMSSLRSEDTAMYYCTRRGYDGVDKWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 227 heavy chain DNA GAAGTGCATCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAACTTTGCCATGTCTTGGGTTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATTAGTACTGGTGGTACTTACACCTACTATCCAGACAGTGTGAAGGGTCGATTCACCATCTCCAGAGACAATGCCAAGAAAACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAGGACACGGCCATGTATTACTGTACAAGACGGGGGTACGACGGCGTGGACAAATGGGGCCAAGGCACCACTCTCACAGTCTCCTCAgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttccccgagcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggacgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctgccagccccaatcgaaa agacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctaccccagcgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 228 LCDR1 (combined) KSGQSLLDSDGKTYLN SEQ ID NO: 229 LCDR2 (combined) LVSKLDS SEQ ID NO: 230 LCDR3 (combined) WQGTHFPQT SEQ ID NO: 231 LCDR1 (Kabat) KSGQSLLDSDGKTYLN SEQ ID NO: 232 LCDR2 (Kabat) LVSKLDS SEQ ID NO: 233 LCDR3 (Kabat) WQGTHFPQT SEQ ID NO: 234 LCDR1 (Chothia) GQSLLDSDGKTY SEQ ID NO: 235 LCDR2 (Chothia) LVS SEQ ID NO: 236 LCDR3 (Chothia) GTHFPQ SEQ ID NO: 237 LCDR1 (IMGT) QSLLDSDGKTY SEQ ID NO: 238 LCDR2 (IMGT) LVS SEQ ID NO: 239 LCDR3 (IMGT) WQGTHFPQT SEQ ID NO: 240 VL DVVMTQTPLTLSVTIGQPASISCKSGQSLLDSDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGTKLEIK SEQ ID NO: 241 DNA VL gatgttgtgatgacccagactccactcactttgtcggttaccattggacaaccagcctccatctcttgcaagtcaggtcagagcctcttagatagtgatggaaagacatatttgaattggtttttacagaggccaggccagtctccaaagcgcctaatctatctggtgtctaaactggactctggagtccctgacaggttcactggcagtggatcagggacagatttcacactgaaaatcagcagagtggaggctgaggatttgggagtttattattgctggcaaggtacacattttcctcagacgttcggtggaggcaccaagctggaaatcaaa SEQ ID NO: 242 light chain DVVMTQTPLTLSVTIGQPASISCKSGQSLLDSDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQRGSSKDSTYSLSSTLVNKYGL SEQ ID NO: 243 light chain DNA GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAGGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTTTTACAGAGGCCAGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAACTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTACACATTTTCCTCAGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAcgtacggtggccgctcccagcgtgttcatcttcccccccagcgacgagcagctgaagagtggcaccgccagcgtggtgtgcctgctgaacaacttctacccccgggaggccaaggtgcagtggaaggtggacaacgccctgcagagcggcaacagccaggagagcgtcaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcataaggtgtacgcctgcgaggtgacccaccagggcctgtccagccccgtgaccaagagcttcaacaggggcgagtgc Original 674J13, hIgG1, CysMab SEQ ID NO: 244 HCDR1 (combined) GYSITSGYSWH SEQ ID NO: 245 HCDR2 (combined) HIHSSGSTNYNPSLKS SEQ ID NO: 246 HCDR3 (combined) GGVQAFAY SEQ ID NO: 247 HDDR1 (Kabat) SGYSWH SEQ ID NO: 248 HDDR2 (Kabat) HIHSSGSTNYNPSLKS SEQ ID NO: 249 HDDR3 (Kabat) GGVQAFAY SEQ ID NO: 250 HCDR1 (Chothia) GYSITSGY SEQ ID NO: 251 HCDR2 (Chothia) HSSGS SEQ ID NO: 252 HCDR3 (Chothia) GGVQAFAY SEQ ID NO: 253 HCDR1 (IMGT) GYSITSGYS SEQ ID NO: 254 HDDR2 (IMGT) IHSSGST SEQ ID NO: 255 HDDR3 (IMGT) ARGGVQAFAY SEQ ID NO: 256 vh DVQLQESGPDLVKPSQSLSLTCTVTGYSITSGYSWHWIRQFPGNKLEWMAHIHSSGSTNYNPSLKSRISIIRDTSKNLFFLQLNSVTTEDTATYYCARGGVQAFAYWGQGTLVTVSA SEQ ID NO: 257 DNA VH GATGTGCAGCTTCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTCAGTCACTTTCACTCACCTGCACTGTCACTGGCTACTCCATCACCAGTGGTTATAGCTGGCACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGCCCACATCCACTCCAGTGGTAGCACTAACTACAACCCATCTCTCAAAAGTCGCATCTCTATCATTCGAGACACATCCAAGAACCTGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGAGGGGGGGTACAGGCCTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA SEQ ID NO: 258 heavy chain
(CysMab, mutations underlined) DVQLQESGPDLVKPSQSLSLTCTVTGYSITSGYSWHWIRQFPGNKLEWMAHIHSSGSTNYNPSLKSRISIIRDTSKNLFFLQLNSVTTEDTATYYCARGGVQAFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 259 heavy chain DNA GATGTGCAGCTTCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTCAGTCACTTTCACTCACCTGCACTGTCACTGGCTACTCCATCACCAGTGGTTATAGCTGGCACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGCCCACATCCACTCCAGTGGTAGCACTAACTACAACCCATCTCTCAAAAGTCGCATCTCTATCATTCGAGACACATCCAAGAACCTGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGAGGGGGGGTACAGGCCTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttcccctgtcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggacgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctgccagccccaatcgaaa agacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctacccctgtgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 260 LCDR1 (combined) SASSSVIYMH SEQ ID NO: 261 LCDR2 (combined) DTSKLAS SEQ ID NO: 262 LCDR3 (combined) QQWSSNPLT SEQ ID NO: 263 LCDR1 (Kabat) SASSSVIYMH SEQ ID NO: 264 LCDR2 (Kabat) DTSKLAS SEQ ID NO: 265 LCDR3 (Kabat) QQWSSNPLT SEQ ID NO: 266 LCDR1 (Chothia) SSSVIY SEQ ID NO: 267 LCDR2 (Chothia) DTS SEQ ID NO: 268 LCDR3 (Chothia) WSSNPL SEQ ID NO: 269 LCDR1 (IMGT) SSVIY SEQ ID NO: 270 LCDR2 (IMGT) DTS SEQ ID NO: 271 LCDR3 (IMGT) QQWSSNPLT SEQ ID NO: 272 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVIYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTTLELK SEQ ID NO: 273 DNA VL CAAATTGTCCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGTTCAAGTGTAATTTACATGCACTGGTACCAGCAGAAGTCAGGCACCTCCCCCAAAAGATGGATTTATGACACATCCAAACTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGTAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCGCTCACGTTCGGTGCTGGGACCACGTTGGAGCTGAAA SEQ ID NO: 274 light chain QIVLTQSPAIMSASPGEKVTMTCSASSSVIYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTTLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACENSFTHQGL SEQ ID NO: 275 light chain DNA CAAATTGTCCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCCAGGGGAGAAGGTCACCATGACCTGCAGTGCCAGTTCAAGTGTAATTTACATGCACTGGTACCAGCAGAAGTCAGGCACCTCCCCCAAAAGATGGATTTATGACACATCCAAACTGGCTTCTGGAGTCCCTGCTCGCTTCAGTGGTAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCAGCAGTGGAGTAGTAACCCGCTCACGTTCGGTGCTGGGACCACGTTGGAGCTGAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 674J13, hIgG1, DAPA, CysMab SEQ ID NO: 276 HCDR1 (combined) GYSITSGYSWH SEQ ID NO: 277 HCDR2 (combined) HIHSSGSTNYNPSLKS SEQ ID NO: 278 HCDR3 (combined) GGVQAFAY SEQ ID NO: 279 HDDR1 (Kabat) SGYSWH SEQ ID NO: 280 HDDR2 (Kabat) HIHSSGSTNYNPSLKS SEQ ID NO: 281 HDDR3 (Kabat) GGVQAFAY SEQ ID NO: 282 HCDR1 (Chothia) GYSITSGY SEQ ID NO: 283 HCDR2 (Chothia) HSSGS SEQ ID NO: 284 HCDR3 (Chothia) GGVQAFAY SEQ ID NO: 285 HCDR1 (IMGT) GYSITSGYS SEQ ID NO: 286 HDDR2 (IMGT) IHSSGST SEQ ID NO: 287 HDDR3 (IMGT) ARGGVQAFAY SEQ ID NO: 288 vh DVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYSWHWIRQHPGKGLEWMAHIHSSGSTNYNPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARGGVQAFAYWGQGTLVTVSS SEQ ID NO: 289 DNA VH GACGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTGAAACCCTCCCAGACCCTGTCCCTGACCTGCACCGTGTCCGGCTACTCCATCACCTCCGGCTACAGCTGGCACTGGATCCGGCAGCACCCCGGCAAGGGCCTGGAATGGATGGCCCACATCCACTCCTCCGGCTCCACCAACTACAACCCCAGCCTGAAGTCCAGAATCACCATCAGCCGGGACACCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCCGCTGACACCGCCGTGTACTACTGTGCCAGAGGCGGCGTGCAGGCCTTCGCTTATTGGGGCCAGGGCACCCTGGTGACAGTGTCCTCC SEQ ID NO: 290 heavy chain
(DAPA, CysMab, mutations are underlined) DVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYSWHWIRQHPGKGLEWMAHIHSSGSTNYNPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARGGVQAFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 291 heavy chain DNA GACGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTGAAACCCTCCCAGACCCTGTCCCTGACCTGCACCGTGTCCGGCTACTCCATCACCTCCGGCTACAGCTGGCACTGGATCCGGCAGCACCCCGGCAAGGGCCTGGAATGGATGGCCCACATCCACTCCTCCGGCTCCACCAACTACAACCCCAGCCTGAAGTCCAGAATCACCATCAGCCGGGACACCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCCGCTGACACCGCCGTGTACTACTGTGCCAGAGGCGGCGTGCAGGCCTTCGCTTATTGGGGCCAGGGCACCCTGGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCTGTCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGGCTGCCCCAATCGAAA AGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCTGTGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 292 LCDR1 (combined) SASSSVIYMH SEQ ID NO: 293 LCDR2 (combined) DTSKLAS SEQ ID NO: 294 LCDR3 (combined) QQWSSNPLT SEQ ID NO: 295 LCDR1 (Kabat) SASSSVIYMH SEQ ID NO: 296 LCDR2 (Kabat) DTSKLAS SEQ ID NO: 297 LCDR3 (Kabat) QQWSSNPLT SEQ ID NO: 298 LCDR1 (Chothia) SSSVIY SEQ ID NO: 299 LCDR2 (Chothia) DTS SEQ ID NO: 300 LCDR3 (Chothia) WSSNPL SEQ ID NO: 301 LCDR1 (IMGT) SSVIY SEQ ID NO: 302 LCDR2 (IMGT) DTS SEQ ID NO: 303 LCDR3 (IMGT) QQWSSNPLT SEQ ID NO: 304 VL EIVLTQSPATLSASPGERVTMSCSASSSVIYMHWYQQKPGQAPRRWIYDTSKLASGVPARFSGSGSGTDYTLTISSMEPEDAAVYYCQQWSSNPLTFGQGTKLEIK SEQ ID NO: 305 DNA VL GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTGCTAGCCCTGGCGAGCGCGTGACAATGTCCTGCTCCGCCTCCTCCTCCGTGATCTACATGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCGGTGGATCTACGATACCTCCAAGCTGGCCTCCGGCGTGCCCGCCAGATTCTCCGGCTCTGGCTCTGGCACCGACTACACCCTGACCATCTCCAGCATGGAACCCGAGGACGCCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCCCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG SEQ ID NO: 306 light chain EIVLTQSPATLSASPGERVTMSCSASSSVIYMHWYQQKPGQAPRRWIYDTSKLASGVPARFSGSGSGTDYTLTISSMEPEDAAVYYCQQWSSNPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS SEQ ID NO: 307 light chain DNA GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTGCTAGCCCTGGCGAGCGCGTGACAATGTCCTGCTCCGCCTCCTCCTCCGTGATCTACATGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCGGTGGATCTACGATACCTCCAAGCTGGCCTCCGGCGTGCCCGCCAGATTCTCCGGCTCTGGCTCTGGCACCGACTACACCCTGACCATCTCCAGCATGGAACCCGAGGACGCCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCCCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 121G12, hIgG1, CysMab SEQ ID NO: 308 HCDR1 (combined) GFTFSTYAMS SEQ ID NO: 309 HCDR2 (combined) TISDAGSYSYYPDNVKG SEQ ID NO: 310 HCDR3 (combined) RGSRYEEYYVMDY SEQ ID NO: 311 HDDR1 (Kabat) TYAMS SEQ ID NO: 312 HDDR2 (Kabat) TISDAGSYSYYPDNVKG SEQ ID NO: 313 HDDR3 (Kabat) RGSRYEEYYVMDY SEQ ID NO: 314 HCDR1 (Chothia) GFTFSTY SEQ ID NO: 315 HCDR2 (Chothia) SDAGSY SEQ ID NO: 316 HCDR3 (Chothia) RGSRYEEYYVMDY SEQ ID NO: 317 HCDR1 (IMGT) GFTFSTYA SEQ ID NO: 318 HDDR2 (IMGT) ISDAGSYS SEQ ID NO: 319 HDDR3 (IMGT) ARRGSRYEEYYVMDY SEQ ID NO: 320 vh EVQLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWVATISDGGSYSYYPDNVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCARRGSRYEEYYVMDYWGQGTSVTVSS SEQ ID NO: 321 DNA VH
(CysMab, mutations underlined) GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGGTCGCAACCATTAGTGATGGTGGTAGTTATTCGTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTATACCTGCAAATGAGCCATCTGAAGTCTGAGGACACAGCCATGTATTACTGTGCAAGACGAGGTAGTAGGTACGAAGAGTACTATGTTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA SEQ ID NO: 322 heavy chain
(CysMab, mutations underlined) EVQLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWVATISDGGSYSYYPDNVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCARRGSRYEEYYVMDYWGQGTSVTVSSastkgpsvfplapsskstsggtaalgclvkdyfp C pvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssslgtqtyicnvnhkpsntkvdkrvepkscdkthtcppcpapellggpsvflfppkpkdtlmisrtpevtcvvvdvshedpevkfnwyvdgvevhnaktkpreeqynstyrvvsvltvlhqdwlngkeykckvsnkalpapiektiskakgqprepqvytlppsreemtknqvsltclvkgfyp C diavewesngqpennykttppvldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspgk SEQ ID NO: 323 heavy chain DNA GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGGTCGCAACCATTAGTGATGGTGGTAGTTATTCGTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTATACCTGCAAATGAGCCATCTGAAGTCTGAGGACACAGCCATGTATTACTGTGCAAGACGAGGTAGTAGGTACGAAGAGTACTATGTTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttcccctgtcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggacgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctgc cagccccaatcgaaaagacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctacccctgtgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 324 LCDR1 (combined) RASQSISNNLH SEQ ID NO: 325 LCDR2 (combined) YASQSIS SEQ ID NO: 326 LCDR3 (combined) QQSSSWLT SEQ ID NO: 327 LCDR1 (Kabat) RASQSISNNLH SEQ ID NO: 328 LCDR2 (Kabat) YASQSIS SEQ ID NO: 329 LCDR3 (Kabat) QQSSSWLT SEQ ID NO: 330 LCDR1 (Chothia) SQSISNN SEQ ID NO: 331 LCDR2 (Chothia) YAS SEQ ID NO: 332 LCDR3 (Chothia) SSSWL SEQ ID NO: 333 LCDR1 (IMGT) QSISNN SEQ ID NO: 334 LCDR2 (IMGT) YAS SEQ ID NO: 335 LCDR3 (IMGT) QQSSSWLT SEQ ID NO: 336 VL DIVLTQSPATLSVTPGDSVSLSCRASQSISNNLHWYQQKSHESPKLLIKYASQSISGIPSRFSGSGSGTDFTLSINSVETEDFGMYFCQQSNSWLTFGAGTKLGLK SEQ ID NO: 337 DNA VL GATATTGTGCTAACTCAGTCTCCAGCCACCCTGTCTGTGACTCCAGGAGATAGCGTCAGTCTTTCCTGCAGGGCCAGCCAAAGTATTAGCAACAACCTACACTGGTATCAACAGAAATCACATGAGTCTCCAAAACTTCTCATCAAGTATGCTTCCCAGTCCATCTCTGGGATCCCCTCCAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACTCTCAGTATCAACAGTGTGGAGACTGAAGATTTTGGAATGTATTTCTGTCAACAGAGTAACAGCTGGCTCACGTTCGGTGCTGGGACCAAGCTGGGGCTGAAA SEQ ID NO: 338 light chain DIVLTQSPATLSVTPGDSVSLSCRASQSISNNLHWYQQQKSHESPKLLIKYASQSISGIPSRFSGSGSGTDFTLSINSVETEDFGMYFCQQSNSWLTFGAGTKLGLKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACESFTHQGLSSPV SEQ ID NO: 339 light chain DNA GATATTGTGCTAACTCAGTCTCCAGCCACCCTGTCTGTGACTCCAGGAGATAGCGTCAGTCTTTCCTGCAGGGCCAGCCAAAGTATTAGCAACAACCTACACTGGTATCAACAGAAATCACATGAGTCTCCAAAACTTCTCATCAAGTATGCTTCCCAGTCCATCTCTGGGATCCCCTCCAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACTCTCAGTATCAACAGTGTGGAGACTGAAGATTTTGGAATGTATTTCTGTCAACAGAGTAACAGCTGGCTCACGTTCGGTGCTGGGACCAAGCTGGGGCTGAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 121G12, hIgG1, DAPA, CysMab SEQ ID NO: 340 HCDR1 (combined) GFTFSTYAMS SEQ ID NO: 341 HCDR2 (combined) TISDAGSYSYYPDNVKG SEQ ID NO: 342 HCDR3 (combined) RGSRYEEYYVMDY SEQ ID NO: 343 HDDR1 (Kabat) TYAMS SEQ ID NO: 344 HDDR2 (Kabat) TISDAGSYSYYPDNVKG SEQ ID NO: 345 HDDR3 (Kabat) RGSRYEEYYVMDY SEQ ID NO: 346 HCDR1 (Chothia) GFTFSTY SEQ ID NO: 347 HCDR2 (Chothia) SDAGSY SEQ ID NO: 348 HCDR3 (Chothia) RGSRYEEYYVMDY SEQ ID NO: 349 HCDR1 (IMGT) GFTFSTYA SEQ ID NO: 350 HDDR2 (IMGT) ISDAGSYS SEQ ID NO: 351 HDDR3 (IMGT) ARRGSRYEEYYVMDY SEQ ID NO: 352 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSTYAMSWVRQAPGKGLEWVATISDAGSYSYYPDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGSRYEEYYVMDYWGQGTTVTVSS SEQ ID NO: 353 DNA VH GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCACCTACGCCATGTCCTGGGTGCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCGACGCCGGCTCCTACTCCTACTACCCCGACAACGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGGCTCCAGATACGAAGAGTACTACGTGATGGACTATTGGGGCCAGGGCACCACCGTGACAGTGTCCTCC SEQ ID NO: 354 heavy chain
(DAPA, CysMab, mutations are underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSTYAMSWVRQAPGKGLEWVATISDAGSYSYYPDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGSRYEEYYVMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 355 heavy chain DNA GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCACCTACGCCATGTCCTGGGTGCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCGACGCCGGCTCCTACTCCTACTACCCCGACAACGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGGCTCCAGATACGAAGAGTACTACGTGATGGACTATTGGGGCCAGGGCACCACCGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCTGTCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGG CTGCCCCAATCGAAAAGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCTGTGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 356 LCDR1 (combined) RASQSISNNLH SEQ ID NO: 357 LCDR2 (combined) YASQSIS SEQ ID NO: 358 LCDR3 (combined) QQSSSWLT SEQ ID NO: 359 LCDR1 (Kabat) RASQSISNNLH SEQ ID NO: 360 LCDR2 (Kabat) YASQSIS SEQ ID NO: 361 LCDR3 (Kabat) QQSSSWLT SEQ ID NO: 362 LCDR1 (Chothia) SQSISNN SEQ ID NO: 363 LCDR2 (Chothia) YAS SEQ ID NO: 364 LCDR3 (Chothia) SSSWL SEQ ID NO: 365 LCDR1 (IMGT) QSISNN SEQ ID NO: 366 LCDR2 (IMGT) YAS SEQ ID NO: 367 LCDR3 (IMGT) QQSSSWLT SEQ ID NO: 368 VL EIVLTQSPATLSVSPGERVTLSCRASQSISNNLHWYQQKPGQAPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSVEPEDFGVYFCQQSSSWLTFGQGTKLEIK SEQ ID NO: 369 DNA VL GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCCGTGTCTCCCGGCGAGAGAGTGACCCTGTCCTGCCGGGCCTCCCAGTCCATCTCCAACAACCTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATTAAGTACGCCTCCCAGAGCATCTCCGGCATCCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACCCTGACCATCTCCAGCGTGGAACCCGAGGACTTCGGCGTGTACTTCTGCCAGCAGTCCTCATCCTGGCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG SEQ ID NO: 370 light chain EIVLTQSPATLSVSPGERVTLSCRASQSISNNLHWYQQKPGQAPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSVEPEDFGVYFCQQSSSWLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACENSFTHQGL SEQ ID NO: 371 light chain DNA GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCCGTGTCTCCCGGCGAGAGAGTGACCCTGTCCTGCCGGGCCTCCCAGTCCATCTCCAACAACCTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATTAAGTACGCCTCCCAGAGCATCTCCGGCATCCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACCCTGACCATCTCCAGCGTGGAACCCGAGGACTTCGGCGTGTACTTCTGCCAGCAGTCCTCATCCTGGCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 506E15, hIgG1, CysMab SEQ ID NO: 372 HCDR1 (combined) GFTFSSYAMS SEQ ID NO: 373 HCDR2 (combined) TISSGGSFTYYPDSVKG SEQ ID NO: 374 HCDR3 (combined) RASTVVGTDFDV SEQ ID NO: 375 HDDR1 (Kabat) SYAMS SEQ ID NO: 376 HDDR2 (Kabat) TISSGGSFTYYPDSVKG SEQ ID NO: 377 HDDR3 (Kabat) RASTVVGTDFDV SEQ ID NO: 378 HCDR1 (Chothia) GFTFSSY SEQ ID NO: 379 HCDR2 (Chothia) SSGGSF SEQ ID NO: 380 HCDR3 (Chothia) RASTVVGTDFDV SEQ ID NO: 381 HCDR1 (IMGT) GFTFSSYA SEQ ID NO: 382 HDDR2 (IMGT) ISSGGSFT SEQ ID NO: 383 HDDR3 (IMGT) ARRASTVVGTDFDV SEQ ID NO: 384 vh EVMLVESGGGLVKPGGSLKLSCAASGFTFSSYAMSWIRQTPEKRLEWVATISSGGSFTYYPDSVKGRFTISRDNVKNTLYLQMSSLRSEDTAMYYCARRASTVVGTDFDVWGAGTTVTVSS SEQ ID NO: 385 DNA VH GAAGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATGCCATGTCTTGGATTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATCAGTAGTGGTGGTAGTTTCACCTACTATCCAGACAGTGTGAAGGGGCGATTCACCATTTCTAGAGACAATGTCAAGAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAAGACACGGCCATGTATTACTGTGCAAGACGGGCTTCTACGGTAGTAGGTACGGACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 386 heavy chain
(CysMab, mutations underlined) EVMLVESGGGLVKPGGSLKLSCAASGFTFSSYAMSWIRQTPEKRLEWVATISSGGSFTYYPDSVKGRFTISRDNVKNTLYLQMSSLRSEDTAMYYCARRASTVVGTDFDVWGAGTTVTVSSastkgpsvfplapsskstsggtaalgclvkdyfp C pvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssslgtqtyicnvnhkpsntkvdkrvepkscdkthtcppcpapellggpsvflfppkpkdtlmisrtpevtcvvvdvshedpevkfnwyvdgvevhnaktkpreeqynstyrvvsvltvlhqdwlngkeykckvsnkalpapiektiskakgqprepqvytlppsreemtknqvsltclvkgfyp C diavewesngqpennykttppvldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspgk SEQ ID NO: 387 heavy chain DNA GAAGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATGCCATGTCTTGGATTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATCAGTAGTGGTGGTAGTTTCACCTACTATCCAGACAGTGTGAAGGGGCGATTCACCATTTCTAGAGACAATGTCAAGAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAAGACACGGCCATGTATTACTGTGCAAGACGGGCTTCTACGGTAGTAGGTACGGACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCAgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttcccctgtcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggacgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctgccag ccccaatcgaaaagacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctacccctgtgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 388 LCDR1 (combined) RASQDIGSSLN SEQ ID NO: 389 LCDR2 (combined) ATSSLDS SEQ ID NO: 390 LCDR3 (combined) LQYASSPPT SEQ ID NO: 391 LCDR1 (Kabat) RASQDIGSSLN SEQ ID NO: 392 LCDR2 (Kabat) ATSSLDS SEQ ID NO: 393 LCDR3 (Kabat) LQYASSPPT SEQ ID NO: 394 LCDR1 (Chothia) SQDIGSS SEQ ID NO: 395 LCDR2 (Chothia) ATS SEQ ID NO: 396 LCDR3 (Chothia) YASSPP SEQ ID NO: 397 LCDR1 (IMGT) QDIGSS SEQ ID NO: 398 LCDR2 (IMGT) ATS SEQ ID NO: 399 LCDR3 (IMGT) LQYASSPPT SEQ ID NO: 400 VL DIQMTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSGSRSGSDYSLTISSLESEDFVVYYCLQYASSPPTFGGGTKLEIK SEQ ID NO: 401 DNA VL GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTATGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGGCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGTCTATTACTGTCTACAATATGCTAGTTCGCCTCCGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA SEQ ID NO: 402 light chain DIQMTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSGSRSGSDYSLTISSLESEDFVVYYCLQYASSPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKHKSSVY SEQ ID NO: 403 light chain DNA GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTATGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGGCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGTCTATTACTGTCTACAATATGCTAGTTCGCCTCCGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 506E15, hIgG1, DAPA, CysMab SEQ ID NO: 404 HCDR1 (combined) GFTFSSYAMS SEQ ID NO: 405 HCDR2 (combined) TISSGGSFTYYPDSVKG SEQ ID NO: 406 HCDR3 (combined) RASTVVGTDFDV SEQ ID NO: 407 HDDR1 (Kabat) SYAMS SEQ ID NO: 408 HDDR2 (Kabat) TISSGGSFTYYPDSVKG SEQ ID NO: 409 HDDR3 (Kabat) RASTVVGTDFDV SEQ ID NO: 410 HCDR1 (Chothia) GFTFSSY SEQ ID NO: 411 HCDR2 (Chothia) SSGGSF SEQ ID NO: 412 HCDR3 (Chothia) RASTVVGTDFDV SEQ ID NO: 413 HCDR1 (IMGT) GFTFSSYA SEQ ID NO: 414 HDDR2 (IMGT) ISSGGSFT SEQ ID NO: 415 HDDR3 (IMGT) ARRASTVVGTDFDV SEQ ID NO: 416 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYAMSWIRQAPGKGLEWVATISSGGSFTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRASTVVGTDFDVWGQGTTVTVSS SEQ ID NO:417 DNA VH GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCAGCTACGCCATGTCCTGGATCCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCTCCGGCGGCAGCTTCACCTACTACCCCGACAGCGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGCCTCCACCGTCGTGGGAACCGACTTCGATGTGTGGGGCCAGGGCACCACCGTGACAGTGTCCTCC SEQ ID NO: 418 heavy chain
(DAPA, CysMab, mutations are underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYAMSWIRQAPGKGLEWVATISSGGSFTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRASTVVGTDFDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 419 heavy chain DNA GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCAGCTACGCCATGTCCTGGATCCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCTCCGGCGGCAGCTTCACCTACTACCCCGACAGCGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGCCTCCACCGTCGTGGGAACCGACTTCGATGTGTGGGGCCAGGGCACCACCGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCTGTCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGGCTG CCCCAATCGAAAAGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCTGTGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 420 LCDR1 (combined) RASQDIGSSLN SEQ ID NO: 421 LCDR2 (combined) ATSSLDS SEQ ID NO: 422 LCDR3 (combined) LQYASSPPT SEQ ID NO: 423 LCDR1 (Kabat) RASQDIGSSLN SEQ ID NO: 424 LCDR2 (Kabat) ATSSLDS SEQ ID NO: 425 LCDR3 (Kabat) LQYASSPPT SEQ ID NO: 426 LCDR1 (Chothia) SQDIGSS SEQ ID NO:427 LCDR2 (Chothia) ATS SEQ ID NO: 428 LCDR3 (Chothia) YASSPP SEQ ID NO: 429 LCDR1 (IMGT) QDIGSS SEQ ID NO: 430 LCDR2 (IMGT) ATS SEQ ID NO:431 LCDR3 (IMGT) LQYASSPPT SEQ ID NO: 432 VL DIQMTQSPSSLSASVGDRVTLTCRASQDIGSSLNWLQQKPGKAIKRLIYATSSLDSGVPSRFSGSRSGTDYTLTISSLQPEDFVVYYCLQYASSPPTFGGGTKLEIK SEQ ID NO:433 DNA VL GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCCGCCTCCGTGGGCGATAGAGTGACCCTGACCTGCCGGGCCTCCCAGGACATCGGCTCCTCCCTGAACTGGCTGCAGCAGAAGCCCGGCAAGGCCATCAAGCGGCTGATCTACGCCACCTCCTCCCTGGACTCCGGCGTGCCCTCCCGGTTCTCCGGCTCTAGATCCGGCACCGACTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGTGGTGTACTACTGCCTGCAGTACGCCTCCAGCCCCCCCACCTTTGGCGGAGGCACCAAGCTGGAAATCAAG SEQ ID NO: 434 light chain DIQMTQSPSSLSASVGDRVTLTCRASQDIGSSLNWLQQKPGKAIKRLIYATSSLDSGVPSRFSGSRSGTDYTLTISSLQPEDFVVYYCLQYASSPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 435 light chain DNA GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCCGCCTCCGTGGGCGATAGAGTGACCCTGACCTGCCGGGCCTCCCAGGACATCGGCTCCTCCCTGAACTGGCTGCAGCAGAAGCCCGGCAAGGCCATCAAGCGGCTGATCTACGCCACCTCCTCCCTGGACTCCGGCGTGCCCTCCCGGTTCTCCGGCTCTAGATCCGGCACCGACTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGTGGTGTACTACTGCCTGCAGTACGCCTCCAGCCCCCCCACCTTTGGCGGAGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Original 684E12, hIgG1, CysMab SEQ ID NO: 436 HCDR1 (combined) GFTFSNFAMS SEQ ID NO:437 HCDR2 (combined) TISTGGTYTYYPDSVKG SEQ ID NO: 438 HCDR3 (combined) RGYDGVDK SEQ ID NO: 439 HDDR1 (Kabat) SNFAMS SEQ ID NO: 440 HDDR2 (Kabat) TISTGGTYTYYPDSVKG SEQ ID NO:441 HDDR3 (Kabat) RGYDGVDK SEQ ID NO:442 HCDR1 (Chothia) GFTFSNF SEQ ID NO:443 HCDR2 (Chothia) STGGTY SEQ ID NO:444 HCDR3 (Chothia) RGYDGVDK SEQ ID NO: 445 HCDR1 (IMGT) GFTFSNFA SEQ ID NO: 446 HDDR2 (IMGT) ISTGGTYT SEQ ID NO:447 HDDR3 (IMGT) TRRGYDGVDK SEQ ID NO: 448 vh EVHLVESGGGLVKPGGSLKLSCAASGFTFSNFAMSWVRQTPEKRLEWVATISTGGTYTYYPDSVKGRFTISRDNAKKTLYLQMSSLRSEDTAMYYCTRRGYDGVDKWGQGTTLTVSS SEQ ID NO:449 DNA VH GAAGTGCATCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAACTTTGCCATGTCTTGGGTTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATTAGTACTGGTGGTACTTACACCTACTATCCAGACAGTGTGAAGGGTCGATTCACCATCTCCAGAGACAATGCCAAGAAAACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAGGACACGGCCATGTATTACTGTACAAGACGGGGGTACGACGGCGTGGACAAATGGGGCCAAGGCACCACTCTCACAGTCTCCTCA SEQ ID NO: 450 heavy chain
(CysMab, mutations underlined) EVHLVESGGGLVKPGGSLKLSCAASGFTFSNFAMSWVRQTPEKRLEWVATISTGGTYTYYPDSVKGRFTISRDNAKKTLYLQMSSLRSEDTAMYYCTRRGYDGVDKWGQGTTLTVSSastkgpsvfplapsskstsggtaalgclvkdyfp C pvtvswnsgaltsgvhtfpavlqssglyslssvvtvpssslgtqtyicnvnhkpsntkvdkrvepkscdkthtcppcpapellggpsvflfppkpkdtlmisrtpevtcvvvdvshedpevkfnwyvdgvevhnaktkpreeqynstyrvvsvltvlhqdwlngkeykckvsnkalpapiektiskakgqprepqvytlppsreemtknqvsltclvkgfyp C diavewesngqpennykttppvldsdgsfflyskltvdksrwqqgnvfscsvmhealhnhytqkslslspgk SEQ ID NO:451 heavy chain DNA
(CysMab, mutations underlined) GAAGTGCATCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAACTTTGCCATGTCTTGGGTTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATTAGTACTGGTGGTACTTACACCTACTATCCAGACAGTGTGAAGGGTCGATTCACCATCTCCAGAGACAATGCCAAGAAAACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAGGACACGGCCATGTATTACTGTACAAGACGGGGGTACGACGGCGTGGACAAATGGGGCCAAGGCACCACTCTCACAGTCTCCTCAgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttcccctgtcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggacgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctgccagccccaatcgaaa agacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctacccctgtgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 452 LCDR1 (combined) KSGQSLLDSDGKTYLN SEQ ID NO: 453 LCDR2 (combined) LVSKLDS SEQ ID NO: 454 LCDR3 (combined) WQGTHFPQT SEQ ID NO: 455 LCDR1 (Kabat) KSGQSLLDSDGKTYLN SEQ ID NO:456 LCDR2 (Kabat) LVSKLDS SEQ ID NO:457 LCDR3 (Kabat) WQGTHFPQT SEQ ID NO: 458 LCDR1 (Chothia) GQSLLDSDGKTY SEQ ID NO:459 LCDR2 (Chothia) LVS SEQ ID NO: 460 LCDR3 (Chothia) GTHFPQ SEQ ID NO:461 LCDR1 (IMGT) QSLLDSDGKTY SEQ ID NO:462 LCDR2 (IMGT) LVS SEQ ID NO:463 LCDR3 (IMGT) WQGTHFPQT SEQ ID NO: 464 VL DVVMTQTPLTLSVTIGQPASISCKSGQSLLDSDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGTKLEIK SEQ ID NO: 465 DNA VL gatgttgtgatgacccagactccactcactttgtcggttaccattggacaaccagcctccatctcttgcaagtcaggtcagagcctcttagatagtgatggaaagacatatttgaattggtttttacagaggccaggccagtctccaaagcgcctaatctatctggtgtctaaactggactctggagtccctgacaggttcactggcagtggatcagggacagatttcacactgaaaatcagcagagtggaggctgaggatttgggagtttattattgctggcaaggtacacattttcctcagacgttcggtggaggcaccaagctggaaatcaaa SEQ ID NO:466 light chain DVVMTQTPLTLSVTIGQPASISCKSGQSLLDSDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQRGSSKDSTYSLSSTLVNKYGL SEQ ID NO:467 light chain DNA GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAGGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTTTTACAGAGGCCAGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAACTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTACACATTTTCCTCAGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAcgtacggtggccgctcccagcgtgttcatcttcccccccagcgacgagcagctgaagagtggcaccgccagcgtggtgtgcctgctgaacaacttctacccccgggaggccaaggtgcagtggaaggtggacaacgccctgcagagcggcaacagccaggagagcgtcaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcataaggtgtacgcctgcgaggtgacccaccagggcctgtccagccccgtgaccaagagcttcaacaggggcgagtgc Original 684E12, hIgG1, DAPA, CysMab SEQ ID NO:468 HCDR1 (combined) GFTFSNFAMS SEQ ID NO:469 HCDR2 (combined) TISTGGTYTYYPDSVKG SEQ ID NO: 470 HCDR3 (combined) RGYDGVDK SEQ ID NO:471 HDDR1 (Kabat) SNFAMS SEQ ID NO:472 HDDR2 (Kabat) TISTGGTYTYYPDSVKG SEQ ID NO:473 HDDR3 (Kabat) RGYDGVDK SEQ ID NO: 474 HCDR1 (Chothia) GFTFSNF SEQ ID NO: 475 HCDR2 (Chothia) STGGTY SEQ ID NO:476 HCDR3 (Chothia) RGYDGVDK SEQ ID NO:477 HCDR1 (IMGT) GFTFSNFA SEQ ID NO: 478 HDDR2 (IMGT) ISTGGTYT SEQ ID NO: 479 HDDR3 (IMGT) TRRGYDGVDK SEQ ID NO: 480 vh EVHLVESGGGLVKPGGSLKLSCAASGFTFSNFAMSWVRQTPEKRLEWVATISTGGTYTYYPDSVKGRFTISRDNAKKTLYLQMSSLRSEDTAMYYCTRRGYDGVDKWGQGTTLTVSS SEQ ID NO: 481 DNA VH GAAGTGCATCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAACTTTGCCATGTCTTGGGTTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATTAGTACTGGTGGTACTTACACCTACTATCCAGACAGTGTGAAGGGTCGATTCACCATCTCCAGAGACAATGCCAAGAAAACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAGGACACGGCCATGTATTACTGTACAAGACGGGGGTACGACGGCGTGGACAAATGGGGCCAAGGCACCACTCTCACAGTCTCCTCA SEQ ID NO: 482 heavy chain
(DAPA, CysMab, mutations are underlined) EVHLVESGGGLVKPGGSLKLSCAASGFTFSNFAMSWVRQTPEKRLEWVATISTGGTYTYYPDSVKGRFTISRDNAKKTLYLQMSSLRSEDTAMYYCTRRGYDGVDKWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 483 heavy chain DNA GAAGTGCATCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAACTTTGCCATGTCTTGGGTTCGCCAGACTCCGGAGAAGAGACTGGAGTGGGTCGCAACCATTAGTACTGGTGGTACTTACACCTACTATCCAGACAGTGTGAAGGGTCGATTCACCATCTCCAGAGACAATGCCAAGAAAACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAGGACACGGCCATGTATTACTGTACAAGACGGGGGTACGACGGCGTGGACAAATGGGGCCAAGGCACCACTCTCACAGTCTCCTCAgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttcccctgtcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggccgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctggctgccccaatcgaaa agacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctacccctgtgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 484 LCDR1 (combined) KSGQSLLDSDGKTYLN SEQ ID NO: 485 LCDR2 (combined) LVSKLDS SEQ ID NO:486 LCDR3 (combined) WQGTHFPQT SEQ ID NO:487 LCDR1 (Kabat) KSGQSLLDSDGKTYLN SEQ ID NO: 488 LCDR2 (Kabat) LVSKLDS SEQ ID NO: 489 LCDR3 (Kabat) WQGTHFPQT SEQ ID NO: 490 LCDR1 (Chothia) GQSLLDSDGKTY SEQ ID NO: 491 LCDR2 (Chothia) LVS SEQ ID NO: 492 LCDR3 (Chothia) GTHFPQ SEQ ID NO: 493 LCDR1 (IMGT) QSLLDSDGKTY SEQ ID NO: 494 LCDR2 (IMGT) LVS SEQ ID NO: 495 LCDR3 (IMGT) WQGTHFPQT SEQ ID NO: 496 VL DVVMTQTPLTLSVTIGQPASISCKSGQSLLDSDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGTKLEIK SEQ ID NO:497 DNA VL gatgttgtgatgacccagactccactcactttgtcggttaccattggacaaccagcctccatctcttgcaagtcaggtcagagcctcttagatagtgatggaaagacatatttgaattggtttttacagaggccaggccagtctccaaagcgcctaatctatctggtgtctaaactggactctggagtccctgacaggttcactggcagtggatcagggacagatttcacactgaaaatcagcagagtggaggctgaggatttgggagtttattattgctggcaaggtacacattttcctcagacgttcggtggaggcaccaagctggaaatcaaa SEQ ID NO: 498 light chain DVVMTQTPLTLSVTIGQPASISCKSGQSLLDSDGKTYLNWFLQRPGQSPKRLIYLVSKLDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPQTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQRGSSKDSTYSLSSTLVNKYGL SEQ ID NO: 499 light chain DNA GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAGGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTTTTACAGAGGCCAGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAACTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTACACATTTTCCTCAGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAcgtacggtggccgctcccagcgtgttcatcttcccccccagcgacgagcagctgaagagtggcaccgccagcgtggtgtgcctgctgaacaacttctacccccgggaggccaaggtgcagtggaaggtggacaacgccctgcagagcggcaacagccaggagagcgtcaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcataaggtgtacgcctgcgaggtgacccaccagggcctgtccagccccgtgaccaagagcttcaacaggggcgagtgc Humanized 674J13, hIgG1, DAPA SEQ ID NO: 500 HCDR1 (combined) GYSITSGYSWH SEQ ID NO: 501 HCDR2 (combined) HIHSSGSTNYNPSLKS SEQ ID NO: 502 HCDR3 (combined) GGVQAFAY SEQ ID NO: 503 HDDR1 (Kabat) SGYSWH SEQ ID NO: 504 HDDR2 (Kabat) HIHSSGSTNYNPSLKS SEQ ID NO: 505 HDDR3 (Kabat) GGVQAFAY SEQ ID NO: 506 HCDR1 (Chothia) GYSITSGY SEQ ID NO: 507 HCDR2 (Chothia) HSSGS SEQ ID NO: 508 HCDR3 (Chothia) GGVQAFAY SEQ ID NO: 509 HCDR1 (IMGT) GYSITSGYS SEQ ID NO: 510 HDDR2 (IMGT) IHSSGST SEQ ID NO: 511 HDDR3 (IMGT) ARGGVQAFAY SEQ ID NO: 512 vh DVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYSWHWIRQHPGKGLEWMAHIHSSGSTNYNPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARGGVQAFAYWGQGTLVTVSS SEQ ID NO: 513 DNA VH GACGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTGAAACCCTCCCAGACCCTGTCCCTGACCTGCACCGTGTCCGGCTACTCCATCACCTCCGGCTACAGCTGGCACTGGATCCGGCAGCACCCCGGCAAGGGCCTGGAATGGATGGCCCACATCCACTCCTCCGGCTCCACCAACTACAACCCCAGCCTGAAGTCCAGAATCACCATCAGCCGGGACACCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCCGCTGACACCGCCGTGTACTACTGTGCCAGAGGCGGCGTGCAGGCCTTCGCTTATTGGGGCCAGGGCACCCTGGTGACAGTGTCCTCC SEQ ID NO: 514 heavy chain
(DAPA, mutations underlined) DVQLQESGPGLVKPSQTLSLTCTVSGYSITSGYSWHWIRQHPGKGLEWMAHIHSSGSTNYNPSLKSRITISRDTSKNQFSLKLSSVTAADTAVYYCARGGVQAFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV a VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL a APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 515 heavy chain DNA GACGTGCAGCTGCAGGAATCTGGCCCTGGCCTGGTGAAACCCTCCCAGACCCTGTCCCTGACCTGCACCGTGTCCGGCTACTCCATCACCTCCGGCTACAGCTGGCACTGGATCCGGCAGCACCCCGGCAAGGGCCTGGAATGGATGGCCCACATCCACTCCTCCGGCTCCACCAACTACAACCCCAGCCTGAAGTCCAGAATCACCATCAGCCGGGACACCTCCAAGAACCAGTTCTCCCTGAAGCTGTCCTCCGTGACCGCCGCTGACACCGCCGTGTACTACTGTGCCAGAGGCGGCGTGCAGGCCTTCGCTTATTGGGGCCAGGGCACCCTGGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGGCTGCCCCAATCGAAA AGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 516 LCDR1 (combined) SASSSVIYMH SEQ ID NO: 517 LCDR2 (combined) DTSKLAS SEQ ID NO: 518 LCDR3 (combined) QQWSSNPLT SEQ ID NO: 519 LCDR1 (Kabat) SASSSVIYMH SEQ ID NO: 520 LCDR2 (Kabat) DTSKLAS SEQ ID NO: 521 LCDR3 (Kabat) QQWSSNPLT SEQ ID NO: 522 LCDR1 (Chothia) SSSVIY SEQ ID NO: 523 LCDR2 (Chothia) DTS SEQ ID NO: 524 LCDR3 (Chothia) WSSNPL SEQ ID NO: 525 LCDR1 (IMGT) SSVIY SEQ ID NO: 526 LCDR2 (IMGT) DTS SEQ ID NO: 527 LCDR3 (IMGT) QQWSSNPLT SEQ ID NO: 528 VL EIVLTQSPATLSASPGERVTMSCSASSSVIYMHWYQQKPGQAPRRWIYDTSKLASGVPARFSGSGSGTDYTLTISSMEPEDAAVYYCQQWSSNPLTFGQGTKLEIK SEQ ID NO: 529 DNA VL GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTGCTAGCCCTGGCGAGCGCGTGACAATGTCCTGCTCCGCCTCCTCCTCCGTGATCTACATGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCGGTGGATCTACGATACCTCCAAGCTGGCCTCCGGCGTGCCCGCCAGATTCTCCGGCTCTGGCTCTGGCACCGACTACACCCTGACCATCTCCAGCATGGAACCCGAGGACGCCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCCCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG SEQ ID NO: 530 light chain EIVLTQSPATLSASPGERVTMSCSASSSVIYMHWYQQKPGQAPRRWIYDTSKLASGVPARFSGSGSGTDYTLTISSMEPEDAAVYYCQQWSSNPLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS SEQ ID NO: 531 light chain DNA GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCTGCTAGCCCTGGCGAGCGCGTGACAATGTCCTGCTCCGCCTCCTCCTCCGTGATCTACATGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCGGTGGATCTACGATACCTCCAAGCTGGCCTCCGGCGTGCCCGCCAGATTCTCCGGCTCTGGCTCTGGCACCGACTACACCCTGACCATCTCCAGCATGGAACCCGAGGACGCCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCCCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Humanized 121G12, hIgG1, DAPA SEQ ID NO: 532 HCDR1 (combined) GFTFSTYAMS SEQ ID NO: 533 HCDR2 (combined) TISDAGSYSYYPDNVKG SEQ ID NO: 534 HCDR3 (combined) RGSRYEEYYVMDY SEQ ID NO: 535 HDDR1 (Kabat) TYAMS SEQ ID NO: 536 HDDR2 (Kabat) TISDAGSYSYYPDNVKG SEQ ID NO: 537 HDDR3 (Kabat) RGSRYEEYYVMDY SEQ ID NO: 538 HCDR1 (Chothia) GFTFSTY SEQ ID NO: 539 HCDR2 (Chothia) SDAGSY SEQ ID NO: 540 HCDR3 (Chothia) RGSRYEEYYVMDY SEQ ID NO: 541 HCDR1 (IMGT) GFTFSTYA SEQ ID NO: 542 HDDR2 (IMGT) ISDAGSYS SEQ ID NO: 543 HDDR3 (IMGT) ARRGSRYEEYYVMDY SEQ ID NO: 544 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSTYAMSWVRQAPGKGLEWVATISDAGSYSYYPDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGSRYEEYYVMDYWGQGTTVTVSS SEQ ID NO: 545 DNA VH GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCACCTACGCCATGTCCTGGGTGCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCGACGCCGGCTCCTACTCCTACTACCCCGACAACGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGGCTCCAGATACGAAGAGTACTACGTGATGGACTATTGGGGCCAGGGCACCACCGTGACAGTGTCCTCC SEQ ID NO: 546 heavy chain
(DAPA, mutations underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSTYAMSWVRQAPGKGLEWVATISDAGSYSYYPDNVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGSRYEEYYVMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV a VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL a APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 547 heavy chain DNA GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCACCTACGCCATGTCCTGGGTGCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCGACGCCGGCTCCTACTCCTACTACCCCGACAACGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGGCTCCAGATACGAAGAGTACTACGTGATGGACTATTGGGGCCAGGGCACCACCGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGG CTGCCCCAATCGAAAAGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 548 LCDR1 (combined) RASQSISNNLH SEQ ID NO: 549 LCDR2 (combined) YASQSIS SEQ ID NO: 550 LCDR3 (combined) QQSSSWLT SEQ ID NO: 551 LCDR1 (Kabat) RASQSISNNLH SEQ ID NO: 552 LCDR2 (Kabat) YASQSIS SEQ ID NO: 553 LCDR3 (Kabat) QQSSSWLT SEQ ID NO: 554 LCDR1 (Chothia) SQSISNN SEQ ID NO: 555 LCDR2 (Chothia) YAS SEQ ID NO: 556 LCDR3 (Chothia) SSSWL SEQ ID NO: 557 LCDR1 (IMGT) QSISNN SEQ ID NO: 558 LCDR2 (IMGT) YAS SEQ ID NO: 559 LCDR3 (IMGT) QQSSSWLT SEQ ID NO: 560 VL EIVLTQSPATLSVSPGERVTLSCRASQSISNNLHWYQQKPGQAPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSVEPEDFGVYFCQQSSSWLTFGQGTKLEIK SEQ ID NO: 561 DNA VL GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCCGTGTCTCCCGGCGAGAGAGTGACCCTGTCCTGCCGGGCCTCCCAGTCCATCTCCAACAACCTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATTAAGTACGCCTCCCAGAGCATCTCCGGCATCCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACCCTGACCATCTCCAGCGTGGAACCCGAGGACTTCGGCGTGTACTTCTGCCAGCAGTCCTCATCCTGGCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAG SEQ ID NO: 562 light chain EIVLTQSPATLSVSPGERVTLSCRASQSISNNLHWYQQKPGQAPRLLIKYASQSISGIPARFSGSGSGTDFTLTISSVEPEDFGVYFCQQSSSWLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACENSFTHQGL SEQ ID NO: 563 light chain DNA GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCCGTGTCTCCCGGCGAGAGAGTGACCCTGTCCTGCCGGGCCTCCCAGTCCATCTCCAACAACCTGCACTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATTAAGTACGCCTCCCAGAGCATCTCCGGCATCCCTGCCAGATTCTCCGGCTCCGGCAGCGGCACCGACTTCACCCTGACCATCTCCAGCGTGGAACCCGAGGACTTCGGCGTGTACTTCTGCCAGCAGTCCTCATCCTGGCTGACCTTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Humanized 506E15, hIgG1, DAPA SEQ ID NO: 564 HCDR1 (combined) GFTFSSYAMS SEQ ID NO: 565 HCDR2 (combined) TISSGGSFTYYPDSVKG SEQ ID NO: 566 HCDR3 (combined) RASTVVGTDFDV SEQ ID NO: 567 HDDR1 (Kabat) SYAMS SEQ ID NO: 568 HDDR2 (Kabat) TISSGGSFTYYPDSVKG SEQ ID NO: 569 HDDR3 (Kabat) RASTVVGTDFDV SEQ ID NO: 570 HCDR1 (Chothia) GFTFSSY SEQ ID NO: 571 HCDR2 (Chothia) SSGGSF SEQ ID NO: 572 HCDR3 (Chothia) RASTVVGTDFDV SEQ ID NO: 573 HCDR1 (IMGT) GFTFSSYA SEQ ID NO: 574 HDDR2 (IMGT) ISSGGSFT SEQ ID NO: 575 HDDR3 (IMGT) ARRASTVVGTDFDV SEQ ID NO: 576 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYAMSWIRQAPGKGLEWVATISSGGSFTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRASTVVGTDFDVWGQGTTVTVSS SEQ ID NO: 577 DNA VH GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCAGCTACGCCATGTCCTGGATCCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCTCCGGCGGCAGCTTCACCTACTACCCCGACAGCGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGCCTCCACCGTCGTGGGAACCGACTTCGATGTGTGGGGCCAGGGCACCACCGTGACAGTGTCCTCC SEQ ID NO: 578 heavy chain
(DAPA, mutations underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYAMSWIRQAPGKGLEWVATISSGGSFTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRASTVVGTDFDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV a VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL a APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 579 heavy chain DNA GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCAGCTACGCCATGTCCTGGATCCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGGCCACCATCTCCTCCGGCGGCAGCTTCACCTACTACCCCGACAGCGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGCCTCCACCGTCGTGGGAACCGACTTCGATGTGTGGGGCCAGGGCACCACCGTGACAGTGTCCTCCGCTAGCACCAAGGGCCCAAGTGTGTTTCCCCTGGCCCCCAGCAGCAAGTCTACTTCCGGCGGAACTGCTGCCCTGGGTTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACAGTGTCCTGGAACTCTGGGGCTCTGACTTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCTCTCTGGGAACCCAGACCTATATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCTCCAGAACTGCTGGGAGGGCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGTCCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTCTCCAACAAGGCCCTGGCTG CCCCAATCGAAAAGACAATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCAGCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCAAG SEQ ID NO: 580 LCDR1 (combined) RASQDIGSSLN SEQ ID NO: 581 LCDR2 (combined) ATSSLDS SEQ ID NO: 582 LCDR3 (combined) LQYASSPPT SEQ ID NO: 583 LCDR1 (Kabat) RASQDIGSSLN SEQ ID NO: 584 LCDR2 (Kabat) ATSSLDS SEQ ID NO: 585 LCDR3 (Kabat) LQYASSPPT SEQ ID NO: 586 LCDR1 (Chothia) SQDIGSS SEQ ID NO: 587 LCDR2 (Chothia) ATS SEQ ID NO: 588 LCDR3 (Chothia) YASSPP SEQ ID NO: 589 LCDR1 (IMGT) QDIGSS SEQ ID NO: 590 LCDR2 (IMGT) ATS SEQ ID NO: 591 LCDR3 (IMGT) LQYASSPPT SEQ ID NO: 592 VL DIQMTQSPSSLSASVGDRVTLTCRASQDIGSSLNWLQQKPGKAIKRLIYATSSLDSGVPSRFSGSRSGTDYTLTISSLQPEDFVVYYCLQYASSPPTFGGGTKLEIK SEQ ID NO: 593 DNA VL GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCCGCCTCCGTGGGCGATAGAGTGACCCTGACCTGCCGGGCCTCCCAGGACATCGGCTCCTCCCTGAACTGGCTGCAGCAGAAGCCCGGCAAGGCCATCAAGCGGCTGATCTACGCCACCTCCTCCCTGGACTCCGGCGTGCCCTCCCGGTTCTCCGGCTCTAGATCCGGCACCGACTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGTGGTGTACTACTGCCTGCAGTACGCCTCCAGCCCCCCCACCTTTGGCGGAGGCACCAAGCTGGAAATCAAG SEQ ID NO: 594 light chain DIQMTQSPSSLSASVGDRVTLTCRASQDIGSSLNWLQQKPGKAIKRLIYATSSLDSGVPSRFSGSRSGTDYTLTISSLQPEDFVVYYCLQYASSPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 595 light chain DNA GACATCCAGATGACCCAGTCCCCCTCCAGCCTGTCCGCCTCCGTGGGCGATAGAGTGACCCTGACCTGCCGGGCCTCCCAGGACATCGGCTCCTCCCTGAACTGGCTGCAGCAGAAGCCCGGCAAGGCCATCAAGCGGCTGATCTACGCCACCTCCTCCCTGGACTCCGGCGTGCCCTCCCGGTTCTCCGGCTCTAGATCCGGCACCGACTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGTGGTGTACTACTGCCTGCAGTACGCCTCCAGCCCCCCCACCTTTGGCGGAGGCACCAAGCTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGTGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC Humanized 684E12, hIgG1, DAPA, CysMab SEQ ID NO: 596 HCDR1 (combined) GFTFSNFAMS SEQ ID NO: 597 HCDR2 (combined) TISTGGTYTYYPDSVKG SEQ ID NO: 598 HCDR3 (combined) RGYSGVDK SEQ ID NO: 599 HDDR1 (Kabat) SNFAMS SEQ ID NO: 600 HDDR2 (Kabat) TISTGGTYTYYPDSVKG SEQ ID NO: 601 HDDR3 (Kabat) RGYSGVDK SEQ ID NO: 602 HCDR1 (Chothia) GFTFSNF SEQ ID NO: 603 HCDR2 (Chothia) STGGTY SEQ ID NO: 604 HCDR3 (Chothia) RGYSGVDK SEQ ID NO: 605 HCDR1 (IMGT) GFTFSNFA SEQ ID NO: 606 HDDR2 (IMGT) ISTGGTYT SEQ ID NO: 607 HDDR3 (IMGT) TRRGYSGVDK SEQ ID NO: 608 vh EVQLVESGGGLVKPGGSLRLSCAASGFTFSNFAMSWVRQAPGKGLEWVSTISTGGTYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGYSGVDKWGQGTTVTVSS SEQ ID NO: 609 DNA VH GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCAACTTCGCCATGTCCTGGGTGCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGTCCACCATCTCCACCGGCGGCACCTACACCTACTACCCCGACAGCGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGGCTACTCAGGCGTGGACAAATGGGGCCAGGGCACCACCGTGACAGTGTCCTCC SEQ ID NO: 610 heavy chain
(DAPA, CysMab, mutations are underlined) EVQLVESGGGLVKPGGSLRLSCAASGFTFSNFAMSWVRQAPGKGLEWVSTISTGGTYTYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGYSGVDKWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFP C PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVV A VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKAL A APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP C DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 611 heavy chain DNA GAGGTGCAGCTGGTGGAATCTGGCGGAGGCCTGGTGAAACCCGGCGGATCCCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCAACTTCGCCATGTCCTGGGTGCGGCAGGCTCCCGGCAAGGGCCTGGAATGGGTGTCCACCATCTCCACCGGCGGCACCTACACCTACTACCCCGACAGCGTGAAGGGCAGATTCACCATCAGCCGGGACAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAGACGGGGCTACTCAGGCGTGGACAAATGGGGCCAGGGCACCACCGTGACAGTGTCCTCCgctagcaccaagggcccaagtgtgtttcccctggcccccagcagcaagtctacttccggcggaactgctgccctgggttgcctggtgaaggactacttcccctgtcccgtgacagtgtcctggaactctggggctctgacttccggcgtgcacaccttccccgccgtgctgcagagcagcggcctgtacagcctgagcagcgtggtgacagtgccctccagctctctgggaacccagacctatatctgcaacgtgaaccacaagcccagcaacaccaaggtggacaagagagtggagcccaagagctgcgacaagacccacacctgccccccctgcccagctccagaactgctgggagggccttccgtgttcctgttcccccccaagcccaaggacaccctgatgatcagcaggacccccgaggtgacctgcgtggtggtggccgtgtcccacgaggacccagaggtgaagttcaactggtacgtggacggcgtggaggtgcacaacgccaagaccaagcccagagaggagcagtacaacagcacctacagggtggtgtccgtgctgaccgtgctgcaccaggactggctgaacggcaaagaatacaagtgcaaagtctccaacaaggccctggctgccccaatcgaaa agacaatcagcaaggccaagggccagccacgggagccccaggtgtacaccctgccccccagccgggaggagatgaccaagaaccaggtgtccctgacctgtctggtgaagggcttctacccctgtgatatcgccgtggagtgggagagcaacggccagcccgagaacaactacaagaccacccccccagtgctggacagcgacggcagcttcttcctgtacagcaagctgaccgtggacaagtccaggtggcagcagggcaacgtgttcagctgcagcgtgatgcacgaggccctgcacaaccactacacccagaagtccctgagcctgagccccggcaag SEQ ID NO: 612 LCDR1 (combined) KSGQSLLDSTGKTYLN SEQ ID NO: 613 LCDR2 (combined) LVSKLDS SEQ ID NO: 614 LCDR3 (combined) WQGTHFPQT SEQ ID NO: 615 LCDR1 (Kabat) KSGQSLLDSTGKTYLN SEQ ID NO: 616 LCDR2 (Kabat) LVSKLDS SEQ ID NO: 617 LCDR3 (Kabat) WQGTHFPQT SEQ ID NO: 618 LCDR1 (Chothia) GQSLLDSTGKTY SEQ ID NO: 619 LCDR2 (Chothia) LVS SEQ ID NO: 620 LCDR3 (Chothia) GTHFPQ SEQ ID NO: 621 LCDR1 (IMGT) QSLLDSTGKTY SEQ ID NO: 622 LCDR2 (IMGT) LVS SEQ ID NO: 623 LCDR3 (IMGT) WQGTHFPQT SEQ ID NO: 624 VL DVVMTQSPLSLPVTLGQPASISCKSGQSLLDSTGKTYLNWFLQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPQTFGGGTKLEIK SEQ ID NO: 625 DNA VL GACGTGGTGATGACCCAGTCCCCCCTGTCCCTGCCTGTGACCCTGGGCCAGCCTGCCTCCATCTCCTGCAAGTCCGGCCAGTCCCTGCTGGACTCCACTGGCAAGACCTACCTGAACTGGTTCCTGCAGCGGCCTGGCCAGTCCCCTCGGCGGCTGATCTACCTGGTGTCCAAGCTGGACAGCGGCGTGCCCGACAGATTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAAGCCGAGGACGTGGGCGTGTACTACTGCTGGCAGGGCACCCACTTCCCCCAGACCTTCGGCGGAGGCACCAAGCTGGAAATCAAG SEQ ID NO: 626 light chain DVVMTQSPLSLPVTLGQPASISCKSGQSLLDSTGKTYLNWFLQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPQTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESTKVTEQDSKDSPVSLSSTLVSSKADYEKHKVDNALQSGNSQESTKVTEQDSKDSPVSFTYSLSSTLVSSKADYEKHKVYYCACE SEQ ID NO: 627 light chain DNA GACGTGGTGATGACCCAGTCCCCCCTGTCCCTGCCTGTGACCCTGGGCCAGCCTGCCTCCATCTCCTGCAAGTCCGGCCAGTCCCTGCTGGACTCCACTGGCAAGACCTACCTGAACTGGTTCCTGCAGCGGCCTGGCCAGTCCCCTCGGCGGCTGATCTACCTGGTGTCCAAGCTGGACAGCGGCGTGCCCGACAGATTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGAAGATCTCCCGGGTGGAAGCCGAGGACGTGGGCGTGTACTACTGCTGGCAGGGCACCCACTTCCCCCAGACCTTCGGCGGAGGCACCAAGCTGGAAATCAAGcgtacggtggccgctcccagcgtgttcatcttcccccccagcgacgagcagctgaagagtggcaccgccagcgtggtgtgcctgctgaacaacttctacccccgggaggccaaggtgcagtggaaggtggacaacgccctgcagagcggcaacagccaggagagcgtcaccgagcaggacagcaaggactccacctacagcctgagcagcaccctgaccctgagcaaggccgactacgagaagcataaggtgtacgcctgcgaggtgacccaccagggcctgtccagccccgtgaccaagagcttcaacaggggcgagtgc

Example 2 In vitro assessment of antibody-induced ADCC activity

[00374] The ability of candidate antibodies to mediate ADCC was assessed using a surrogate ADCC reporter assay. JVM2 cells expressing CCR7 and CD20 were used as target cells. JVM2 cells were washed and resuspended at 8×10 ⁴ cells/ml cells/ml. The effector cells in this assay were the Jurkat cell line stably expressing CD16V158 and the NFAT-dependent luciferase reporter (Jurkat-V158); luciferase expression is a substitute for canonical ADCC signaling via CD16. Briefly, Jurkat-V158 cells grown in suspension culture were centrifuged to remove depleted medium, the pellet was resuspended in assay medium and adjusted to 1.6×10 ⁶ cells/ml cells/ml. Mixed equal volumes of effector cells and target cells to obtain the main mixture of cells, in which the ratio of target cells to effector cells is 1:5 or 1:20. The antibody titer was diluted in assay medium at a final highest concentration of 50 μg/ml per assay well. 12.5 μl of the Ab solution was added to a 384-well round bottom plate and then 12.5 μl of the basic cell mixture was added. The antibody and cells were thoroughly mixed by pipetting and incubated for 4 hours at 37°C in 5% CO ₂ . After incubation, 15 µl of Bright Glo substrate (Promega #G7572) was added to each well and shaken for 5 min at RT at 1050 rpm. The luminescent signal was read on an Envision plate reader (Perkin Elmer).

[00375] An antibody targeting CD20 was included as a control and showed significant NFAT signaling as measured by luciferase activity. Similarly, candidate antibodies to CCR7 induced significant ADCC activity (Figure 1).

[00376] The table below summarizes typical results for antibodies of various formats included in the ADCC assay using JVM2 and Jurkat-V158 cells.

Table 5. ADCC activity in non-humanized and humanized anti-CCR7 antibodies

ADCC activity; IC50 (nM) 121G12, non-humanized, CysMab 0.077 506E15, inhumanized, CysMab 0.131 674J13, inhumanized, CysMab 1.09 684E12, inhumanized, CysMab 2.63 121G12, humanized, CysMab Present, but insufficient for curve fitting 506E15, humanized, CysMab 0.054 674J13, humanized, CysMab Present, but insufficient for curve fitting ADCC control, anti-CD20 antibody 0.132

[00377] An ADCC assay was performed with the non-humanized 506E15 antibody as a representative anti-CCR7 antibody capable of mediating ADCC on various cell lines with different numbers of CCR7 receptors to determine the minimum receptor density required for ADCC activity and whether there is an appropriate margin of safety compared to normal CCR7+ T cells. The table below summarizes some of the data.

Table 6 ADCC activity of non-humanized 506E15 antibody

cell line Number of CCR7 receptors 506E15, IC50 (nM) 506E15 (maximum RLU) JVM2 66,500 0.046 5840 MOTN-1 64,600 0.077 5387 DEL 62,000 0.074 6480 CMLT-1 29,000 0.011 4400 SR786 27,800 0.011 4067 PEER 8,700 0.038 3360 ALL-SIL 6,200 0.008 3720 Jurkat V158 3,400 0.004 3267 DND-41 1,700 0.002 3733 Normal T cells < 1.5 thousand cells depletion in vivo n.a.

[00378] The data show that even very low levels of CCR7 receptors, comparable to levels of CCR7 receptors on normal T cells, were sufficient to provide significant ADCC activity. ADCC was also evaluated using a viability assay when NK cells were co-cultured with CCR7+ cancer cells and similar results were obtained (not discussed here).

[00379] This is consistent with the observation described below that T cell depletion was observed in vivo and found to be based on an ADCC mechanism. These results demonstrate that the ADCC mechanism makes CCR7 inapplicable from a security point of view. Therefore, all candidates were switched to the silent Fc (DAPA) format to improve overall target safety.

[00380] The in vitro ADCC reporter assay was repeated and confirmed the absence of ADCC activity in non-humanized anti-CCR7 antibodies containing DAPA mutated Fc variants (Figure 2).

Example 3 Biochemical Characterization of Antibodies

Affinity of Antibodies to CCR7 for CCR7

[00381] The affinity of various antibodies and ADCs for CCR7 and its species orthologues was determined using FACS. Purified IgGs were titrated to determine EC50 values for binding to CCR7 expressed on the cell surface.

[00382] For this, CCR7-positive cells were harvested (adherent cells were separated with accutase), washed twice with FACS buffer (PBS/3% FCS/0.02% sodium azide) and diluted to about 2×10 ⁶ cells/ml in the FACS buffer. All subsequent steps were performed on ice to prevent internalization of the receptor. 100 μl of cell suspension/well was transferred to 96-well U-bottom plates (Falcon). 2x10 ⁵ cells/well were incubated with a serial dilution of the anti-CCR7 antibody of interest, with the antibody concentration varying by several orders of magnitude, from the highest 100 nM, for 60 minutes at 4° C. with gentle shaking. After incubation, cells were centrifuged (1200 rpm, 2 min, 4°C) and washed three times with FACS buffer. The detection antibody, APC fluorophore-conjugated anti-hFc-gamma antibody (Jackson Immunoresearch), was added at a rate of 1:400 and the samples were incubated for 1 hour on ice in the dark with gentle shaking. After a final wash, the cells were resuspended in 100 µl of FACS buffer containing 0.2 µg/ml DAPI, followed by a flow cytometry device (BD LSRFortessa Cell Analyzer; Cat # 647177). Mean fluorescence intensity (MFI) of live single cells was calculated in Flowjo 10.0.8 and exported to Graphpad Prism6 for EC ₅₀ determination.

[00383] Selectivity was assessed by measuring observed binding affinities to an isogenic pair of cells engineered to overexpress CCR7 as well as cell lines expressing CCR7 paralogs, eg CCR9, CCR6, CXCR4 and CCR8. All anti-CCR7 antibodies bound in a specific manner only to cells expressing CCR7, as shown in Table 7 below.

Table 7. Binding of various anti-CCR7 antibodies to cells expressing CCR7

Observed FACS Binding (at 5 µg/mL) CysMab humanized antibody (with wild-type Fc) 121G12 506E15 674J13 684E12 NIH3T3 cells No binding No binding No binding No binding NIH3T3.hCCR7 Binding Binding Binding Binding Cells PF382
(CCR7-/ CCR9+/ CCR6+) No binding Lack of binding No binding Lack of binding HEK293-CXCR4 No binding No binding Lack of binding No binding CHO-CCR8 Lack of binding No binding Lack of binding Lack of binding

[00384] In a similar experiment, antibodies were tested for cross-reactivity using sets of engineered isogenically matched cell lines (NIH3T3 series) and CD4+ T cells, which were purified from multiple batches of PBMC from healthy donors, as well as cynomolgus monkeys, Wistar rats, and CD- mice. one. All antibodies were found to specifically bind human and cynomolgus monkey CCR7 with similar observed affinities as shown in Tables 8 and 9 below. Only the non-humanized 121G12 antibody is cross-reactive with the rodent molecule.

Table 8. Cross-reactivity of various non-humanized antibodies to CCR7

Observed affinity in FACS; EC ₅₀ (nM) Non-humanized antibody CysMab 121G12 506E15 674J13 684E12 human CD4+ T cells 22-30 4.6-9.7 0.39-1.4 n.d. CD4+ T cells of the cynomolgus macaque 5.8-12 6.6 0.45 n.d. mouse CD4+ T cells 48-51 No binding No binding n.d. Rat CD4+ T cells 18-31 No binding Lack of binding n.d. NIH3T3 with human CCR7 1.3 n.d. 3.6 >30 NIH3T3 with CCR7 cynomolgus monkey 1.3 n.d. n.d. Binding NIH3T3 with CCR7 mouse 2.5 No binding No binding No binding NIH3T3 with rat CCR7 2.4 No binding No binding No binding Cancer cells Jeko-1 (CCR7+) 1.5 n.d. n.d. 12

Table 9. Cross-reactivity of various humanized anti-CCR7 antibodies

Observed affinity in FACS; EC ₅₀ (nM) Humanized Antibody CysMab 121G12 506E15 674J13 684E12 human CD4+ T cells 30-34 7.8-11 0.87-2.7 6.7 CD4+ T cells of the cynomolgus macaque 12 7 0.5 n.d. NIH3T3 with human CCR7 one n.d. n.d. n.d. NIH3T3 with CCR7 cynomolgus monkey 0.9 n.d. n.d. n.d.

[00385] To determine the effect of receptor density on observed affinity, and hence the contribution of avidity to antibody-to-cell binding, FACS titrimetric experiments were performed on cancer cell lines expressing human CCR7 at various expression levels and normal CCR7-positive T cells derived from from PBMC. Receptor quantitation was performed by FACS using microspheres from Bangs Laboratories as counting standards and following the manufacturer's instructions. Illustrative results are shown in Table 10 below.

Table 10. Contribution of avidity to observed affinity as a function of receptor density

Observed affinity in FACS; EC ₅₀ (nM) Humanized antibody CysMab.DAPA Density of CCR7 receptors 121G12 506E15 674J13 684E12 NIH3T3.hCCR7 cells >1,000,000 2.5 1.8 0.64 n.d. Cancer cells DEL ~100,000 1.78 3.09 0.47 n.d. human CD4+ T cells <2,000 ~30 ~10 ~2 ~7

[00386] All anti-CCR7 antibodies showed a significant contribution of avidity to the observed affinity, and the binding strength decreased depending on the density of the receptors. In particular, 121G12 shows significantly weaker binding on low CCR7-expressing normal CD4+ T cells compared to DEL cancer cells, representative of the cells for which it is indicated.

[00387] The relatively low affinity of particular 121G12, which showed the strongest avidity effect of any anti-CCR7 antibody, is optimal for using the difference in receptor density between normal and cancer cells as a method to shift antibody binding towards cancer cells.

Binding to recombinant hCCR7 in ELISA

[00388] Binding and affinity were also evaluated in an ELISA-based assay using recombinant CCR7 (Origene, No. TP306614). Maxisorp™ 384-well plates (Thermo Nunc) were coated with 3.5 μg/ml recombinant CCR7 diluted in PBS. After blocking with 3% BSA (bovine serum albumin) in PBS for 1 hour at room temperature, washing plates 3x with PBS-T (0.01% Tween 20 in PBS), serial dilutions of primary antibodies were added and incubated for 1 hour at room temperature. Plates were washed again and bound antibodies were detected by incubation with 1:5000 horseradish peroxidase conjugated anti-hFc-gamma antibody (HRP; Jackson Immunoresearch, Cat # 115-035-098) for 1 hour at room temperature followed by washing using PBS-T and then adding SureBlue peroxidase substrate (KPL, #52-00-03). After 15 min, absorbance was recorded at 650 nm and analyzed in GraphPad Prism6.

[00389] All anti-CCR7 antibodies tested are capable of binding recombinant hCCR7 (Table 11; Figure 3).

Table 11 Binding affinity of humanized anti-CCR7 antibodies to recombinant hCCR7

Humanized Antibody CysMab Affinity, Kd (nM) 121G12.DAPA 0.677 506E15.DAPA 5.731 674J13.DAPA 0.006

Binding with double FabGraft

[00390] FabGraft ELISA was performed to assess binding to the minimum extent epitope that contains the N-terminus and EC2 from CCR7. Briefly, Maxisorp™ 384-well plates (Thermo Nunc) were coated with 5 μg/ml FabGraft. In all other respects, the general instructions for the ELISA protocol were followed as described above. All anti-CCR7 antibodies are capable of binding to double FabGrafts as shown in Table 12 below.

Table 12. Binding affinity of humanized anti-CCR7 antibodies to FabGraft

Humanized Antibody CysMab ELISA; Kd (nM) 121G12.DAPA 0.023 506E15.DAPA 0.025 674J13.DAPA 0.023

Assessment of pH-dependence in ELISA using VLP

[00391] CCL19 bound to CCR7 is known to be internalized as a receptor-ligand complex. However, while CCR7 is recycled back to the cell surface, CCL19 is sorted into the lysosome for degradation, showing the opposite fate of endocytotic CCR7 and its ligand (Otero et al., J Immunol 2006; 177:2314-2323). For an anti-CCR7 ADC to be effective, it is preferred that the antibody behave similarly to the ligand, eg, be rapidly internalized but not recycled back along with CCR7. To achieve this, the authors of the present invention ensured the selection of pH-dependent antibodies that will exhibit weaker binding to CCR7 under low pH conditions.

[00392] To assess the pH-dependence of antibodies to CCR7, ELISA was performed using virus-like particles (VLP) expressing CCR7. Briefly, Maxisorp™ 384-well plates (Thermo Nunc) were coated with 25 μg/ml VLP. Primary antibodies were incubated either in pH 5.8 buffer (1:1; dH ₂ O:0.1 M citrate buffer, 150 mM NaCl) or pH 7.4 buffer. In all other respects, the general instructions for the ELISA protocol were followed as described above.

[00393] Using a range of humanized variants for each candidate, a comparison of antibody binding at neutral (7.4) and acidic (5.8) pH showed that all CCR7 candidate antibodies have improved affinity at pH 7.4 (Table 13 ). The objects below were selected based on their highest pH dependence, among other features.

Table 13. pH-dependence of binding of humanized anti-CCR7 antibodies to VLPs expressing CCR7

ELISA; Kd (nM) Humanized Antibody CysMab Buffer pH 7.4 Buffer pH 5.8 fold change (pH 5.8/7.4) 121G12 0.1411 0.5207 four 506E15 0.0571 0.4942 9 674J13 0.0162 0.1657 ten 684E12 0.0374 0.5041 13

b-arrestin assay

[00394] To determine the functionality of anti-CCR7 antibodies, a β-arrestin assay was performed using DiscoverX's PathHunter Pulse Detection Kit (No. 93-0247) in either agonist mode to assess agonist function or antagonist mode to assess antagonist function.

[00395] In the case of the CHO-flpin-hCCR7 agonist regimen (a cell line developed by DiscoverX expressing hCCR7 labeled with ProLink, β-arrestin-EA) were plated at 8×10 ⁴ cells/well in 20 μl/well of growth medium (medium F -12 Ham/Glutamax; Invitrogen+10% FBS+0.5 mg/mL G418+0.2 mg/mL hygromycin B; Invitrogen+5 µg/mL blasticidin; Gibco) with 100 ng/mL Dox in 384-well plates covered with metal lids, were incubated at 37°C, 5% CO ₂ overnight. The following day, serial dilutions of 5x working solution in 1x assay buffer (20 mM HEPES/0.1% BSA/1x HBSS, pH 7.4) were performed on test antibodies or a positive control using hCCL19 ligand (R&D, 361/MI- 025/CF). 5 µl of 5x working solution of antibodies or ligand was added to each well, fast centrifuged and incubated at 37°C/5% CO ₂ for 2 hours. After incubation, 25 µl of detection reagent was added to each well, incubated at room temperature in the dark with shaking for 20 min. The luminescence signal was then measured on an Envision instrument to determine enzyme activity. Finally, enzyme activity was analyzed using Excel.

[00396] In antagonist mode, CHO-flpin-hCCR7 cells were seeded as described above. The following day, a 6x working solution (0.5 μM x 6 = 3.0 μM) in 1x assay buffer was prepared for each test antibody, either a MAB197 positive control (reference antibody; R&D antagonist ligand) or hIgG negative control. 5 µl of 6x working solution of antibodies or controls was added to each well, fast centrifuged and incubated at 37°C/5% CO ₂ for 30 min. During incubation, serial dilutions of 6x hCCL19 working solution in 1x assay buffer were performed. After incubation, 5 μl of 6x hCCL19 working solution was added to each well, fast centrifuged and incubated at 37°C/5% CO ₂ for 90 min. After incubation, 25 μl of detection reagent was added to each well, incubated at room temperature in the dark with shaking for 20 min. The luminescence signal was then measured on an Envision instrument to determine enzyme activity and analyzed using Excel.

[00397] None of the original anti-CCR7 antibodies showed activity in the agonist model (Figure 4A). However, when analyzed in antagonist format, 506E15 and 121G12 were identified as strong antagonists, eg, antibodies blocking ligand binding (Figures 4B, 4C). 674J12 is a neutral antibody that does not block ligand binding. 684E12 is a weak antagonist.

Competition binding analysis in FACS using a ligand

[00398] To confirm that the antibody competes with the CCR7 ligand, a FACS analysis was performed in the presence of an excess concentration of the ligand. FACS analysis was performed as described above. Some changes were made to the protocol, for example, CCL19 was maintained at a constant concentration of 1 μM, while the concentration of the primary antibody, which was simultaneously applied to DEL cells, varied in the range of several orders of magnitude, starting with the highest concentration of 100 nM. After a 30 min incubation period in ice-cold FACS buffer, cells were washed, a secondary anti-hFc antibody conjugated to PE was applied for 15 min, and MFI was determined as described above.

[00399] Figure 5 shows that humanized CysMab.DAPA 674J13 was not affected by the presence of excess CCL19, confirming its neutral functionality. The binding affinity of humanized CysMab.DAPA 121G12 and 506E15 was indeed strongly affected by the presence of excess ligand.

The ability to internalize in various cell lines characterized by a range of receptor densities

[00400] Another aspect of effecting ADC to CCR7 is to ensure optimal use of the differential distribution of CCR7 expression. As a representative of normal cells expressing CCR7, CD4+ T cells were isolated from PBMCs of healthy donors. In addition, a variety of CCR7-positive cancer cell lines exhibiting a range of receptor densities were selected. The present inventors specifically selected antibodies with a weaker observed FACS binding affinity for CCR7+ T cells relative to CCR7+ cancer cells. In addition, this document describes the pHrodo assay, in which a low pH-activated fluorophore label on anti-CCR7 antibodies is used to evaluate how fluorescence-measured antibody uptake by cells correlates with receptor density. It is preferable to minimize antibody uptake by normal cells in order to maximize the therapeutic window.

[00401] Briefly, labeling of an anti-CCR7 antibody in CysMab format with maleimide-pHrodo (ThermoFisher) was performed following the manufacturer's instructions to obtain objects with DAR=4 (ratio of drug, eg, fluorophore, to antibody). FACS analysis was performed as described above. Some changes were made to the protocol, for example, to allow for internalization, the primary antibody was incubated at 5 μg/ml with cells at 37°C in culture medium for 6 h, then washed with ice-cold FACS buffer containing sodium azide to termination of the reaction.

[00402] Table 14 below summarizes the internalization capacity of three antibodies using a range of cell lines. An untargeted pHrodo-labeled antibody was used as a control, and background noise signal was found to be up to 400 MFI, eg, off-target-mediated capture of the antibody conjugate. The data show that all three anti-CCR7 antibodies require levels of the CCR7 receptor in the range that is typical for most CCR7+ cancer cell lines, e.g. over 20,000 receptors, to be efficiently internalized and accumulate the conjugated substance, while at the same time not falling into normal CD4+ T cells.

Table 14. Internalization capacity of various anti-CCR7 antibodies

cell line Number of receptors pHrodo (6 h, MFI) Humanized 121G12.CysMab.
DAPA Humanized 506E15.CysMab.
DAPA Humanized 617J13.CysMab.
DAPA CD4+ T cells 2000 278 429 402 JVM3 8717 148 238 146 CMLT1 19583 301 465 561 Jeko-1 28852 450 908 1389 Mec2 50840 1340 3089 3004 L1236 61602 1237 3126 5824 MOTN1 84200 1026 2899 2539 DEL 110685 1913 3058 3777 MJ 111121 1649 3870 3259 KE97 152093 1567 2723 4416 L540 167549 6836 16117 11800

Epitope-specific sorting using the Octet Red96 system

[00403] Epitope-specific sorting of the original antibodies to hCCR7 was performed using the Octet Red96 system (ForteBio, USA), which performs biolayer interferometry (BLI). The CCR7 immunogenic backbone was biotinylated with AviTag™ using BirA biotin ligase according to the manufacturer's recommendations (Avidity, LLC, US cat. no. BirA500). The biotinylated immunogenic backbone was loaded at a rate of 1.5 µg/ml onto pre-balanced streptavidin sensors (ForteBio, USA). Then the sensors were transferred into a solution containing 100 nM antibody A in 1X buffer for kinetic analysis (ForteBio, USA). The sensors were washed briefly in 1X kinetic assay buffer and transferred to a second solution containing 100 nM competitor antibody. Binding kinetics were determined from raw data using Octet Red96 system analysis software (version 6.3, ForteBio, USA). Antibodies were tested in all pairwise combinations as both antibody A and antibody competitor.

Table 15 Antibody Sorting Results

Category Antibody one 684E12; MAB197 2 674J13 3 506E15 four 121G12

Epitope mapping using CCR7 mutations

[00404] Additional epitope mapping was performed using cell lines with mutant CCR7. NIH/3T3 cell lines were created expressing mutated human CCR7 variants. Mutations were introduced at specific positions to replace the human CCR7 residue with the corresponding mouse CCR7 residue. The introduced mutations covered D35E, F44Y, L47V, S49F, D198G, R201K, S202N, S204G, Q206D, A207T, M208L, I213V, T214S, E215A and H216Q. Mutant CCR7 plasmid constructs were generated by site-directed mutagenesis and introduced into NIH/3T3 cells to generate stable expression cell lines. The specific binding of candidate antibodies to each cell line with mutant CCR7 was assessed by flow cytometry. Cells were thoroughly rinsed with PBS and treated with accutase (Millipore, no. SCR005) to separate from growth plates and resuspended at approximately 1x10 ⁵ cells/90 μl in 1x FACS buffer (2% FBS+0.1% NaN ₃ in PBS). 10 µl of 10x antibody solution in FACS buffer was preliminarily added to a 96-well U-bottomed plate and 90 µl of cell suspension was added. Cells were incubated for 30 minutes at 4°C, washed 1x with cold PBS and resuspended in 100 μl of secondary antibody diluted 1:500 in 1x FACS buffer (conjugated with goat allophycocyanin F(ab')2 to human IgG, Fcγ- specific; Jackson Immunoresearch, Cat # 109-136-098). After an additional 15 min incubation at 4° C., cells were washed twice with PBS and resuspended in 100 μl of 1x FACS buffer+4 μg/ml propidium iodide (Life Technologies, Cat # P3566). Geometric mean fluorescence intensity was calculated on live single cells using FlowJo and plotted as % of geometric mean fluorescence intensity at CCR7 WT.

[00405] EC50-based affinities for CCR7 with point mutations showed that all antibodies tested had different binding profiles, indicating different use of the conformational epitope (Figure 6). Of all the antibodies shown, 506E15 has a binding pattern most similar to that of MAB197, e.g., both show a significant dramatic decrease in binding affinity when the F44 or L47 residues at the N-terminus of CCR7 are mutated, and no decrease when M208 or I213 are mutated in the EC2 loop . It appears that 121G12 and 674J13 use a different set of critical contact points within the conformational epitope space as they differ from MAB197 in at least 4 of the 8 point mutations tested.

Example 4 Design and Characterization of Anti-CCR7 Antibody Drug Conjugates

In the section below, µl is used to refer to the microliter. Similarly, µM is used for micromoles/L; and µm is used to refer to a micrometer.

[00406] Example 4A. Preparation of the antibody-drug conjugate 121G12.CysMab.DAPA.MPET.DM4

[00407] Conjugation of purified 121G12.CysMab.DAPA and MPET.DM4

[00408] The starting material was the 121G12.CysMab.DAPA antibody at 127 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 10 mM histidine hydrochloride buffer. To 7.9 ml of antibody (1003 mg) was added 16 ml of 0.5 M sodium phosphate, pH 8 (Teknova S1280), checked for pH >7, then the antibody was absorbed onto 100.3 ml of RMP Resin with Protein A (GE Healthcare 1-223BPO/I) for 25 minutes with gentle swirling at room temperature. Resin loaded at 10 mg Ab/mL layer was washed with 15 layer volumes of 1x PBS buffer (Hyclone SH30256.02) by vacuum filtration through a 0.2 µm filter device (Nalgene 567-0020) placed on neck of the bottle, then resuspended in 100.3 ml 1x PBS to obtain a 50% slurry.

[00409] To the slurry was added 4329 µl of 0.5 M cysteine (Sigma G121-03) formulated in 0.5 M phosphate, pH 8, to which NaOH (Alfa Aesar A16037) was added at a ratio of 13.6 g/L. The slurry was stirred occasionally by rotating the vessel at room temperature for 30 minutes, then washed by vacuum filtration through a 0.2 µm filter device placed on the neck of the bottle with 50 bed volumes of 1x PBS over at least 10 filtration cycles. and additions. The washed resin was resuspended in 100.3 ml of 1x PBS (50% slurry) and 1003 μl of 100 μM CuCl ₂ (Aldrich 751944) (total 500 nM Cu 2 ⁺ ) was added to initiate re-oxidation. Reoxidation of the antibody was tested by removing a 30 µl aliquot of the suspension, adding 1 µl of a 20 mM reference maleimide stock solution (Example 3, page 110 in WO2015/095301), which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuge at 7000 x g for 10 seconds, remove the supernatant, add 60 µl of IgG Thermo elution buffer (Thermo Scientific 21009), centrifuge at 14000 x g for 10 seconds, sample the supernatant and analyze the products by RPLC as follows: 2 -μl sample was injected into a heated (80°C) column 4.6×50 mm Agilent PLRP-S (particles 5 μm, pore size 4000 Å), equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/v water (Millipore TX1280P-1, Burdick and Jackson 407-4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00410] When RPLC analysis indicated that reoxidation was optimal (in this case 60 minutes), 3010 μl of a 20 mM stock solution of MPET.DM4 in DMSO was added and the slurry was gently mixed occasionally by swirling the vessel at room temperature for 30 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00411] The slurry was then transferred to a porous glass filter column (Pierce 7375021), pre-eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (201 ml) was neutralized with 20.1 ml of 0.5 M sodium phosphate, pH 8, then concentrated to 60 ml using centrifuge concentrators (Amicon UFC905024) at 3000× g . The concentrate was then applied to 24 x PD-10 buffer exchange columns (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loading 2.5 ml concentrate and eluting with 3.5 ml 1x PBS as recommended by the manufacturer.

[00412] For stability studies, the material was combined with a batch obtained in an identical manner to provide 2 grams of starting material. The pooled material was extensively dialyzed (Slidealyzer flask, Thermo Scientific 87762) against 10 mM histidine chloride buffer, pH 5 (histidine from JTBaker 2080-05), concentrated to ~30 mg/mL, then sucrose (Millipore 1.00892.1003) was added and Tween 20 (JTBaker 4116-04) to 240 mM and 0.02% (v/v), respectively. For excess material that was not required for stability studies, buffer back exchange was performed with 1x PBS. The samples were divided into aliquots, subjected to rapid freezing with liquid nitrogen and stored at -80°C. Final concentrations were 23.9 mg/ml for material formulated for stability testing and 18.9 mg/ml for material formulated in 1x PBS.

[00413] The analysis of the obtained samples is as follows.

Parameter Stability Sample Sample in 1x PBS Concentration (mg/ml) 23.9 18.9 Pyrogen (EU/ml) 0.1 0.05 % aggregates <1 <1 DAR 3.79 3.79

[00414] Methods of analysis. The concentration was determined based on the fact that for a 10 mg/ml IgG solution, the extinction coefficient at OD280 is 13.7 Pyrogen was determined using a Kinetic QCL assay (Lonza Walkersville 50-650H) with a TECAN Safire plate reader. The percentage of aggregates was determined by analytical size exclusion chromatography on a Shodex KW-G guard column (Thomson Instrument Company, cat. no. 6960955) and a KW-803 column (TIC, cat. no. 6960940), equilibrated with a mobile phase [20 mM Tris, ~pH 7.65 (prepared with 10 mM Tris, pH 7.4, 10 mM Tris, pH 8), 200 mM NaCl, 0.02% sodium azide], obtaining data at 280 nm. For DAR determination, an aliquot of the sample was prepared, which was obtained by diluting the sample to 2 mg/ml in 1x PBS, deglycosylating the sample with PNGase F (obtained in the laboratory) for 10 minutes at 50°C, removing PNGase F by binding to protein A, washing with 1x PBS and eluting with 1% formic acid. The sample was then injected into a 2.1×50 mm PLRP-S column (8 µm particles, pore size 1000 Å) equilibrated with 0.1% formic acid in 20% CH ₃ CN/water (Invitrogen) at a flow rate of 0.5 ml /min The column was washed with 20% CH ₃ CN/in water for 3 minutes, then eluted with a 0.1 minute gradient to 0.1% formic acid in 90% CH ₃ CN/in water, while this concentration was maintained at within 1.9 minutes. Mass spectral data were acquired on an Agilent 1260 instrument and deconvolution was performed using MassHunter Qualitative Analysis software version B.05.00 over the 110-180 kDa range. Peak areas corresponding to different calculated DAR states were weighted according to the DAR of each peak, then summed and the weighted DAR4 peak area was divided by the sum of all weighted peaks to give the DAR value.

[00415] Obtain MPET.DM4 payload linker

[00416] Analytical methods

Unless otherwise indicated, the following HPLC and HPLC/MS methods were used in the Preparation of Intermediates and Examples.

LC/MS analysis was performed on an Agilent 1200sl/6140 system.

Column: Waters Acquity HSS T3 C18, 50×2.0, 1.8 µm

Mobile phase: A) H ₂ O+0.05% TFA; B: acetonitrile+0.035% TFA

Pump operation parameters:

Time A% B% Flow (ml/min) 0 90 ten 0.9 1.35 0 100 0.9 1.36 0 100 0.9 1.95 0 100 0.9 1.96 90 ten 0.9 2.0 90 ten 0.9

Detection: Diode array UV detector at 190nm - 400nm

Scan at MS: 200-1350 amu

ELSD: 60°C

MS options:

Polarity Positive drying gas 12 Spray pressure fifty Drying gas temperature 350 Capillary voltage 3000

[00417] (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6 -dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenecyclotetradecaphan-10,12-dien-4-yl N-(4-(( 2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)ethyl)disulfanyl)-4-methylpentanoyl)-N-methyl-L-alaninate

[00418] Step 1: Preparation of (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl -12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenecyclotetradecaphan-10,12-dien-4-yl N-( 4-((2-aminoethyl)disulfanyl)-4-methylpentanoyl)-N-methyl-L-alaninate

[00419] To DM4 (480 mg, 0.62 mmol) dissolved in PBS buffer (10.5 mL) and anhydrous THF (21 mL) was added 2-(pyridin-2-yldisulfanyl)ethan-1-amine (151 mg, 0.68 mmol) and DIEA (0.27 ml, 1.54 mmol) at room temperature. The reaction mixture was stirred at room temperature for 30 min and concentrated in vacuo. The aqueous residue was diluted with CH ₃ CN (1 ml) and H ₂ O (2 ml) and purified by reverse phase ISCO, eluted with 10-60% acetonitrile in H ₂ O containing 0.05% TFA. Fractions containing the desired product were lyophilized to give the desired product (555 mg, 93% yield). ¹ H NMR (400 MHz, MeOD- d ₄ ) δ ppm 0.83 (s, 3 H) 1.21 (d, J =5.0 Hz, 3 H) 1.25 (s, 3 H) 1, 28 (s, 3 H) 1.30 (d, J =5.0 Hz, 3 H) 1.45-1.55 (m, 3 H) 1.67 (s, 3 H) 1.84-1 .88 (m, 1 H) 1.95-2.01 (m, 1 H) 2.14 (dd, J = 5.0 and 15.0 Hz, 1 H) 2.37-2.43 (m , 1 H) 2.53-2.59 (m, 1 H) 2.64 (dd, J =10.0 and 15.0 Hz, 1 H) 2.82-2.89 (m, 5 H) 2.91 (d, J =10.0 Hz, 1 H) 3.16 (dd, J = 5.0 and 10.0 Hz, 2 H) 3.20 (s, 3 H) 3.23 (d , J =10.0 Hz, 1 H) 3.35 (s, 3 H) 3.55 (d, J =5.0 Hz, 1 H) 3.58 (d, J =10.0 Hz, 1 H) 4.15-4.20 (m, 1 H) 4.64 (dd, J = 5.0 and 10.0 Hz, 1 H) 5.43 (q, J = 5.0 Hz, 2 H ) 5.66 (dd, J = 10.0 and 15.0 Hz, 1 H) ) 6.58 (dd, J = 10.0 and 15.0 Hz, 1 H) 6.65 (d, J = 10.0 Hz, 1 H) 6.66 (s, 1 H) 7.11 (bs, 1H) 7.28 (bs, 1H); MS mass/charge 855.3 (M+H), retention time 0.988 minutes.

[00420] Step 2: Preparation of (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl -12,6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenecyclotetradecaphan-10,12-dien-4-yl N-( 4-((2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)ethyl)disulfanyl)-4-methylpentanoyl)-N-methyl-L-alaninate

[00421] K (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12, 6-dioxo-7-aza-1(6,4)-oxazinana-3(2,3)-oxirana-8(1,3)-benzenecyclotetradecaphan-10,12-dien-4-yl-N-(4- ((2-aminoethyl)disulfanyl)-4-methylpentanoyl)-N-methyl-L-alaninate (555mg, 0.57mmol) dissolved in anhydrous DMSO (7mL) was added 2,5-dioxopyrrolidin-1-yl -3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (171 mg, 0.63 mmol) and DIEA (249 ml, 1.43 mmol) at room temperature. The reaction mixture was stirred at room temperature for 15 minutes and neutralized with TFA. The mixture was cooled to 0°C with an ice bath, followed by the addition of CH ₃ CN (2 ml) and H ₂ O (7 ml), and then purified by reverse phase ISCO, while eluting with 10-70% acetonitrile in H ₂ O containing 0.05% TFA. Fractions containing the desired product were lyophilized to give the desired product (430 mg, 66% yield). ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 0.81 (s, 3 H) 1.23 (s, 3 H) 1.24 (s, 3 H) 1.25 (s, 1 H) 1, 28 (d, J =5.0 Hz, 3 H) 1.31 (d, J = 5.0 Hz, 3 H) 1.43-1.49 (m, 1 H) 1.61 (d, J =15.0 Hz, 1 H) 1.64 (s, 3 H) 1.81-1.87 (m, 1 H) 1.94-2.01 (m, 1 H) 2.19 (dd, J = 5.0 and 15.0 Hz, 1 H) 2.30-2.36 (m, 1 H) 2.54 (t, J = 5.0 Hz, 2 H) 2.61 (dd, J =10.0 and 15.0 Hz, 1 H) 2.70 (t, J =5.0 Hz, 2 H) 2.88 (s, 3 H) 3.00 (d, J =10.0 Hz , 1 H) 3.13 (d, J =10.0 Hz, 1 H) 3.21 (s, 3 H) 3.55 (s, 3 H) 3.45 (q, J = 5.0 Hz , 2 H) 3.49 (d, J =5.0 Hz, 1 H) 3.62 (d, J =10.0 Hz, 1 H) 3.83 (t, J =5.0 Hz, 1 H) 3.98 (s, 3 H) 4.32 (m, 1 H) 4.80 (dd, J = 5.0 and 10.0 Hz, 1 H) 5.28 (d, J = 5, 0 Hz, 1 H) 5.66 (dd, J = 10.0 and 15.0 Hz, 1 H) 6.22 (bs, 1 H) 6.42 (dd, J = 10.0 and 15, 0 Hz, 1 H) 6.50 (s, 1 H) 6.63 (s, 1 H) 6.66 (d, J =10.0 Hz, 1 H) 6.70 (s, 2H) 6, 83(s, 1H); MS mass/charge 988.3 (M+HH ₂ O), retention time 1.145 minutes.

[00422] Example 4B. Preparation of the 121G12.DAPA.sSPDB.DM4 antibody-drug conjugate

[00423] To a stirred solution of 25 mM HEPES buffer, pH 7.6 (3 ml; sterile) and dimethylacetamide (DMAc; 0.12 ml) at 22°C was added a solution (1.695 ml) of 121G12.DAPA; MW ~145546 g/mol; 62 mg (0.426 µmol)) in potassium phosphate buffer (10 mM, pH 6; sterile). Maytansinoid DM4 (2.42 mg (3.101 µmol) dissolved in 0.242 ml DMAc) was added. Sulfo-SPDB linker (0.970 mg (2.386 µmol, corrected for analysis)) dissolved in 0.970 ml DMAc was added. After 18 hours, the reaction mixture was analyzed for completeness reactions using SEC-UV and HPLC.

[00424] The reaction mixture was purified from low molecular weight by-products and buffer exchange was carried out by filtration through Amicon membrane cells; cut-off molecular weight 30 kDa, using 10 mM PBS buffer, pH 7.4 (sterile) for washing. The ultraconcentrates obtained after application of Amicon were pooled and diluted to 10 mg/ml (UV) to obtain 2.9 ml of a solution of the antibody-drug conjugate, 121G12.DAPA.sSPDB-DM4, in 10 mM PBS buffer, pH 7.4 (protein recovery 49%).

[00425] Using SEC-UV, the drug to antibody ratio was determined to be n=3.6 and the monomer purity was 98.7%. Endotoxin levels were 0.14 EU/mg (BET Endosafe test).

[00426] Example 4C. Obtaining an antibody-drug conjugate 684E12.SMCC.DM1

(n=3.6)

[00427] To an antibody solution (stock 684E12) (7.1 mg/mL, 3.4 mL, approx. μl of 4 mM sulfo-SMCC (0.15 mg) in DMA and the mixture was incubated and gently stirred at 4° C. overnight. After incubation, the reaction mixture was purified by desalting on a HiPrep 26/10 desalting column (GE Healthcare) using PBS, pH 7.4 as running buffer and sterilized by filtration. The purified conjugate was analyzed by MALDI-MS and the estimated DAR was 2.6. Analytical SEC indicated that the sample had 3.7% aggregation (or 96.3% monomers) and LAL testing (PTS, Charles River Laboratories) determined an endotoxin value of 0.36 EU/mg.

[00428] Example 4D. Obtaining an antibody-drug conjugate 506E15.AURIX1

The starting material was 506E15.CysMab (Fc WT) at 18.8 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 1x phosphate buffered saline (1x PBS). 1.76 ml of antibody was absorbed onto 3 ml of protein A RMP resin (GE Healthcare 1-223BPO/I) and 240 μl of 0.5 M cysteine (Sigma G121-03) formulated in 0.5 M phosphate was added to the resulting slurry, pH 8 to which NaOH (Alfa Aesar A16037) was added at a ratio of 13.6 g/l. The slurry was stirred occasionally by rotating the vessel at room temperature for 30 minutes, then washed by vacuum filtration through a 0.2 µm filter device placed on the neck of the bottle with 50 bed volumes of 1x PBS over at least 10 filtration cycles. and additions. The washed resin was resuspended in 2 ml 1x PBS (50% slurry) and 60 μl 100 μM CuCl2 (Aldrich 751944) (total 100 nM Cu2 ⁺ ) was added to initiate re-oxidation. After 420 minutes, an additional 90 μl of 100 μM CuCl ₂ was added to further accelerate the re-oxidation. Reoxidation of the antibody was tested by removing a 30 µl aliquot of the suspension, adding 1 µl of a 20 mM reference maleimide stock solution (Example 3, page 110 in WO2015/095301), which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuge at 7000 x g for 10 seconds, remove the supernatant, add 60 µl of Thermo IgG elution buffer (Thermo Scientific 21009), centrifuge at 14000 x g for 10 seconds, sample the supernatant and analyze the products by RPLC as follows: 2 -μl sample was injected into a heated (80°C) column 4.6×50 mm Agilent PLRP-S (particles 5 μm, pore size 4000 Å), equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/v water (Millipore TX1280P-1, Burdick and Jackson 407-4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00429] When RPLC analysis indicated that reoxidation was optimal (in this case 565 minutes), 220 µl of a 20 mM AURIX1 stock solution in DMSO was added and the slurry was occasionally gently mixed by swirling the vessel at room temperature for 70 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00430] The slurry was then transferred to a porous glass filter column (Pierce 7375021), pre-eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (6 ml) was neutralized with 0.6 ml of 0.5 M sodium phosphate, pH 8, concentrated to 2.5 ml using a centrifuge concentrator (Amicon UFC905024) at 3000 × g , applied to a PD-10 buffer exchange column ( GE Healthcare 17-0851-01) equilibrated with 1x PBS, loading 2.5 ml concentrate and eluting with 3.5 ml 1x PBS as recommended by the manufacturer. The yield was 22 mg (66%).

[00431] Example 4E. Obtaining an antibody-drug conjugate 506E15.CysMab.DAPA.AURIX2

The starting material was the 506E15.CysMab.DAPA antibody at 10 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 1x phosphate buffered saline (1x PBS). 2.0 ml of antibody was absorbed onto 2 ml of protein A RMP resin (GE Healthcare 1-223BPO/I) and 160 μl of 0.5 M cysteine (Sigma G121-03) formulated in 0.5 M phosphate was added to the resulting slurry, pH 8 to which NaOH (Alfa Aesar A16037) was added at a ratio of 13.6 g/l. The slurry was stirred occasionally by rotating the vessel at room temperature for 30 minutes, then washed by vacuum filtration through a 0.2 µm filter device placed on the neck of the bottle with 50 bed volumes of 1x PBS over at least 10 filtration cycles. and additions. The washed resin was resuspended in 2 ml 1x PBS (50% slurry) and 10 μl 100 μM CuCl ₂ (Aldrich 751944) (total 250 nM Cu 2 ⁺ ) was added to initiate re-oxidation. Reoxidation of the antibody was tested by removing a 30 µl aliquot of the suspension, adding 1 µl of a 20 mM reference maleimide stock solution (Example 3, page 110 in WO2015/095301), which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuge at 7000 x g for 10 seconds, remove the supernatant, add 60 µl of Thermo IgG elution buffer (Thermo Scientific 21009), centrifuge at 14000 x g for 10 seconds, sample the supernatant and analyze the products by RPLC as follows: 2 -μl sample was injected into a heated (80°C) column 4.6×50 mm Agilent PLRP-S (particles 5 μm, pore size 4000 Å), equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/v water (Millipore TX1280P-1, Burdick and Jackson 407-4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00432] When RPLC analysis indicated that reoxidation was optimal (in this case 295 minutes), 80 μl of a 20 mM AURIX2 stock solution in DMSO was added and the slurry was gently mixed occasionally by swirling the vessel at room temperature for 85 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00433] The slurry was then transferred to a porous glass filter column (Pierce 7375021), pre-eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (4 ml) was neutralized with 0.4 ml 0.5 M sodium phosphate, pH 8, applied to 2 PD-10 buffer exchange columns (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loading 2.5 ml eluate and elute with 3.5 ml 1x PBS as recommended by the manufacturer. The yield was 13.3 mg (67%).

[00434] Example 4F. Obtaining the conjugate of the antibody and drug 674J13.CysMab.AURIX1

[00435] The starting material was 674J13.CysMab (Fc WT) at 9 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 1x phosphate buffered saline (1x PBS). DTT to 200 mM (Invitrogen 15508-013) was added to 57.6 ml of antibody and the solution was incubated for 75 minutes for strong Ab recovery. The recovered Abs were then applied to PD-10 buffer exchange columns (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loading 2.5 ml concentrate and eluting with 3.5 ml 1x PBS as recommended by the manufacturer. The eluates from the PD10 columns were pooled, then reapplied to fresh PD10 columns for more complete removal of DTT. Note that in separate experiments, PD10 columns are more efficient in removing DTT than would be observed using the size exclusion chromatography mechanism alone, provided that the columns are used only once.

[00436] Antibody reoxidation was tested by removing a 30 μl aliquot of the suspension, adding 1 μl of 20 mM stock solution of AURIX1, which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuging at 7000 × g for 10 seconds , removing the supernatant, adding 60 µl of Thermo IgG elution buffer (Thermo Scientific 21009), centrifuging at 14,000 × g for 10 seconds, sampling the supernatant and analyzing the products by RPLC as follows: 2 µl of the sample was injected into a heated (80° C) 4.6×50 mm Agilent PLRP-S column (particles 5 μm, pore size 4000 Å) equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/water (Millipore TX1280P-1, Burdick and Jackson 407-4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00437] When RPLC analysis indicated that reoxidation was optimal (in this case 180 minutes), 290 µl of a 20 mM AURIX1 stock solution in DMSO was added along with 6 ml of protein A RMP resin (GE Healthcare 1-223BPO/I). The slurry was stirred occasionally by rotating the vessel at room temperature for 40 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00438] The slurry was then transferred to a porous glass filter column (Pierce 7375021), pre-eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (11.5 ml) was neutralized with 1.2 ml of 0.5 M sodium phosphate, pH 8, concentrated to 2.5 ml using a centrifuge concentrator (Amicon UFC905024) at 3000 × g , applied to a PD- buffer exchange column. 10 (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loaded with 2.5 ml of concentrate and eluted with 3.5 ml of 1x PBS as recommended by the manufacturer. The yield was 26 mg (41%).

[00439] 4G example. Obtaining the conjugate of the antibody and drug 674J13.CysMab.DAPA.AURIX2

[00440] The starting material was the 674J13.CysMab.DAR4.DAPA antibody at 31.7 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 1x phosphate buffered saline (1x PBS). 9.5 ml of antibody was absorbed into 30.1 ml of protein A RMP resin (GE Healthcare 1-223BPO/I) and 1800 mg of DTT (Invitrogen 15508-013) was added to the resulting slurry for strong Ab recovery (200 mM DTT in total). The slurry was stirred by rotating the vessel at room temperature for 20 minutes, then washed by vacuum filtration through a 0.2 µm filter device placed on the neck of the bottle with 50 bed volumes of 1x PBS over at least 10 filtration cycles and additions. The washed resin was resuspended in 2 ml of 1x PBS and mixed by rotating the vessel at room temperature, with no addition of copper (it accelerated the re-oxidation too much). Reoxidation of the antibody was tested by removing a 30 µl aliquot of the suspension, adding 1 µl of a 20 mM reference maleimide stock solution (Example 3, page 110 in WO2015/095301), which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuge at 7000 x g for 10 seconds, remove the supernatant, add 60 µl of Thermo IgG elution buffer (Thermo Scientific 21009), centrifuge at 14000 x g for 10 seconds, sample the supernatant and analyze the products by RPLC as follows: 2 -μl sample was injected into a heated (80°C) column 4.6×50 mm Agilent PLRP-S (particles 5 μm, pore size 4000 Å), equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/v water (Millipore TX1280P-1, Burdick and Jackson 407-4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00441] When RPLC analysis indicated that reoxidation was optimal (in this case 80 minutes), 903 µl of a 20 mM stock solution of AURIX2 in DMSO was added and the slurry was gently mixed occasionally by swirling the vessel at room temperature for 50 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00442] The slurry was then transferred to a porous glass filter column (Pierce 7375021), pre-eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (60.2 ml) was neutralized with 6.0 ml of 0.5 M sodium phosphate, pH 8, concentrated to 17.5 ml using a centrifuge concentrator (Amicon UFC905024) at 3000 x g , applied to 7 PD buffer exchange columns -10 (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loaded with 2.5 ml of concentrate and eluted with 3.5 ml of 1x PBS as recommended by the manufacturer. The yield was 243 mg (81%)

[00443] Example 4H. Obtaining the conjugate of the antibody and drug 121G12.CysMab.DAPA.AURIX1

[00444] The starting material was the 121G12.CysMab.DAPA antibody at 16.7 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 1x phosphate buffered saline (1x PBS). 3.6 ml of antibody was absorbed into 6 ml of protein A RMP resin (GE Healthcare 1-223BPO/I) and the resulting slurry was stirred by rotating the vessel at room temperature for 125 minutes, then 544 μl of 0.5 M cysteine (Sigma G121 -03) formulated in 0.5 M phosphate, pH 8, to which NaOH (Alfa Aesar A16037) was added at a ratio of 13.6 g/l. The slurry was stirred occasionally by rotating the vessel at room temperature for 60 minutes, then washed by vacuum filtration through a 0.2 µm filter device placed on the neck of the bottle with 50 bed volumes of 1x PBS over at least 10 filtration cycles. and additions. The washed resin was resuspended in 2 ml 1x PBS (50% slurry) and 16 μl 100 μM CuCl ₂ (Aldrich 751944) (total 250 nM Cu 2 ⁺ ) was added to initiate re-oxidation. Antibody reoxidation was tested by removing a 30 µl aliquot of the suspension, adding 1 µl of 20 mM AURIX1 stock solution, which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuging at 7000 × g for 10 seconds, removing the supernatant , adding 60 µl of Thermo IgG elution buffer (Thermo Scientific 21009), centrifugation at 14,000 × g for 10 seconds, sampling the supernatant, and analyzing the products by RPLC as follows: 2 µl sample was injected into a heated (80°C) column 4.6 x 50 mm Agilent PLRP-S (5 µm particles, 4000 Å pore size) equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/water (Millipore TX1280P-1, Burdick and Jackson 407- 4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00445] When RPLC analysis indicated that reoxidation was optimal (in this case 170 minutes), 67 µl of a 20 mM AURIX1 stock solution in DMSO was added and the slurry was gently mixed occasionally by swirling the vessel at room temperature for 120 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00446] The slurry was then transferred to a porous glass filter column (Pierce 7375021), preliminarily eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (15 ml) was neutralized with 1.5 ml 0.5 M sodium phosphate, pH 8, concentrated to 5 ml using a centrifuge concentrator (Amicon UFC905024) at 3000 x g, applied to 2 PD-10 buffer exchange columns (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loading 2.5 ml eluate and eluting with 3.5 ml 1x PBS as recommended by the manufacturer. The yield was 54 mg (92%).

[00447] Example 4I. Obtaining the conjugate of the antibody and drug 121G12.CysMab.AURIX1

The starting material was 121G12.CysMab (Fc WT) at 12.5 mg/mL (for a 10 mg/mL IgG solution, the extinction coefficient at OD280 is 13.7) in 1x phosphate buffered saline (1xPBS). 4.8 ml of antibody was absorbed onto 6 ml of protein A RMP resin (GE Healthcare 1-223BPO/I) and the resulting slurry was stirred by rotating the vessel at room temperature for 125 minutes, then 592 μl of 0.5 M cysteine (Sigma G121 -03) formulated in 0.5 M phosphate, pH 8, to which NaOH (Alfa Aesar A16037) was added at a ratio of 13.6 g/l. The slurry was stirred occasionally by rotating the vessel at room temperature for 60 minutes, then washed by vacuum filtration through a 0.2 µm filter device placed on the neck of the bottle with 50 bed volumes of 1x PBS over at least 10 filtration cycles. and additions. The washed resin was resuspended in 2 ml 1x PBS (50% slurry) and 16 μl 100 μM CuCl ₂ (Aldrich 751944) (total 250 nM Cu 2 ⁺ ) was added to initiate re-oxidation. Antibody reoxidation was tested by removing a 30 µl aliquot of the suspension, adding 1 µl of 20 mM AURIX1 stock solution, which is known to shift the antibody peak in RPLC, mixing for 1 minute, centrifuging at 7000 × g for 10 seconds, removing the supernatant , adding 60 µl of Thermo IgG elution buffer (Thermo Scientific 21009), centrifugation at 14,000 × g for 10 seconds, sampling the supernatant, and analyzing the products by RPLC as follows: 2 µl sample was injected into a heated (80°C) column 4.6 x 50 mm Agilent PLRP-S (5 µm particles, 4000 Å pore size) equilibrated with 0.1% trifluoroacetic acid in 29.5% CH ₃ CN/water (Millipore TX1280P-1, Burdick and Jackson 407- 4) at a flow rate of 1.5 ml/min. The column was eluted with a 5 minute gradient to 44.5% CH ₃ CN/in water, maintained at this concentration for 1.9 minutes and the peaks were determined at 280 nm. The optimal time for conjugation is defined as the time at which the peak of the main product reaches its maximum value, the peaks with later elution are minimized, and the peaks with earlier elution no longer increase.

[00448] When RPLC analysis indicated that reoxidation was optimal (in this case 160 minutes), 67 µl of a 20 mM stock solution of AURIX1 in DMSO was added and the slurry was gently mixed occasionally by swirling the vessel at room temperature for 120 minutes. The slurry was then washed with 20 bed volumes of 1x PBS over at least 10 filtration and addition cycles.

[00449] The slurry was then transferred to a porous glass filter column (Pierce 7375021), pre-eluted with 0.5 bed volumes of IgG Thermo elution buffer (discarded), then eluted with 2 bed volumes of the same buffer. The entire eluate (15 ml) was neutralized with 1.5 ml 0.5 M sodium phosphate, pH 8, concentrated to 5 ml using a centrifuge concentrator (Amicon UFC905024) at 3000 x g, applied to 2 PD-10 buffer exchange columns (GE Healthcare 17-0851-01) equilibrated with 1x PBS, loading 2.5 ml eluate and eluting with 3.5 ml 1x PBS as recommended by the manufacturer. The yield was 52 mg (89%).

Analysis methods. The concentration was determined based on the fact that for a 10 mg/ml IgG solution, the extinction coefficient at OD280 is 13.7 Pyrogen was determined using a Kinetic QCL assay (Lonza Walkersville 50-650H) with a TECAN Safire plate reader. The percentage of aggregates was determined by analytical size exclusion chromatography on a Shodex KW-G guard column (Thomson Instrument Company, cat. no. 6960955) and a KW-803 column (TIC, cat. no. 6960940), equilibrated with a mobile phase [20 mM Tris, ~pH 7.65 (prepared with 10 mM Tris, pH 7.4, 10 mM Tris, pH 8), 200 mM NaCl, 0.02% sodium azide], obtaining data at 280 nm. To determine DAR, an aliquot of the sample was prepared by diluting the sample to 2 mg/ml in 1x PBS, deglycosylating the sample with PNGase F (obtained in the laboratory) for 10 minutes at 50°C, removing PNGase F by protein A binding, washing with using 1x PBS and eluting with 1% formic acid. The sample was reconstituted by adding 1/4 volume 5 M ammonium acetate, pH 5.0 containing 0.5 M TCEP, and incubating at room temperature for 30 minutes. The sample was then injected into a 2.1×50 mm PLRP-S column (8 µm particles, pore size 1000 Å) equilibrated with 0.1% formic acid in 20% CH₃CN/in water (Invitrogen) at 0.5 ml/min. The column was washed with 20% CH₃CN/in water for 3 minutes, then eluted with a 0.1 minute gradient to 0.1% formic acid in 90% CH₃CN/in water, while this concentration was maintained for 1.9 minutes. Mass spectral data were acquired on an Agilent 1260 instrument and deconvolution was performed using MassHunter Qualitative Software version B.05.00 over the 15-60 kDa range. Peak areas corresponding to different calculated DAR states were weighted according to the DAR of each peak, then summed, and the weighted DAR4 peak area was divided by the sum of all weighted peaks to give the DAR value.

The analysis of the obtained sample is as follows.

Parameter 506E15.DAPA.
AURIX2 506E15.
AURIX1 674J13.DAPA.
AURIX2 674J13.
AURIX1 121G12.
AURIX1 121G12.DAPA
AURIX1 Concentration (mg/ml) 1.9 6.7 9 8.4 7.5 7.8 Pyrogen (EU/ml) 0.05 0.42 0.05 <0.5 0.05 0.05 % aggregates <1 <1 2.8 <1 1.2 1.9 DAR 3.80 3.78 HC 3.93
LC 0.03 HC 3.8 3.80 3.80

[00450] Example 4J. Obtaining additional conjugates using other CysMab antibodies

[00451] The methods described in Example 4A were also used to prepare conjugates of MPET.DM4 with other engineered cysteine antibodies.

[00452] The methods are used to prepare P-cadherin-binding Ab.CysMab.MPET.DM4 conjugates using antibodies NOV169N31Q(E152C-S375C), NEG0012(E152C-S375C), NEG0013(E152C-S375C), NEG0016(E152C-S375C ), NEG0064(E152C-S375C), NEG0067(E152C-S375C), NOV169N31Q(K360C(HC)-K107C(LC)), NEG0012(K360C(HC)-K107C(LC)), NEG0013(K360C(HC)-K107C (LC)), NEG0016(K360C(HC)-K107C(LC)), NEG0064(K360C(HC)-K107C(LC)) and NEG0067(K360C(HC)-K107C(LC)), disclosed in PCT Publication No. WO2016/203432.

Example 5 In vitro ADC characterization

[00453] Characteristics of antibody drug conjugates (ADC) were determined using various functional and analytical methods. As assessed by FACS, ADCs retained binding to the CCR7 target protein on cells. For all ADCs, the geometric mean of the fluorescence intensity in the FACS binding assay was within 20% of the value for the unconjugated antibody. Analytical SEC showed that >95% of the ADC material had the desired molecular weight; in cases where this was not observed for the initial reaction products, the use of preparative SEC made it possible to achieve the required specification. The drug to antibody ratio (DAR) was assessed by LCMS of a deglycosylated reconstituted antibody sample, summing the occurrence of different DAR varieties and balancing the number of drug molecules on each DAR variety (e.g. one DAR2 ion counted as 2, one DAR1 ion counted as one). Conjugates containing constant regions containing two cysteine mutations were characterized by at least a DAR of 3.4 or greater and most commonly a DAR of 3.8 or greater. For all registered conjugates, the data were consistent.

Example 6 Cell Proliferation Inhibition/Cell Viability Assay

[00454] Above, we have shown that fluorophore-conjugated variants of all three anti-CCR7 antibodies could lead to internalization of CCR7 and efficient accumulation of the conjugated fluorophore in low pH cell compartments for a range of cell lines. In this example, the present inventors show the ability of an antibody to internalize and accumulate a conjugate within a cell in a mode where the conjugate is a toxic payload.

[00455] In ADC "piggyback" mode (pgADC), the cytotoxic effects of anti-CCR7 antibodies in combination with a payload-conjugated secondary antibody fragment were studied by assessing cell viability after four days of treatment. Three-fold dilutions of CCR7-specific IgGs were prepared and mixed with a constant amount of Fab fragment associated with the payload. The final concentration of the Fab fragment associated with the payload was 0.5 μg/ml. The Fab reagent is a mouse Fc-targeted Fab conjugated to either MMAF or saporin (Advanced Targeting Systems, Fab-Zap). After a 30 min pre-incubation at room temperature, 10 μl/well of the antibody-payload complex was added to 384-well white bottom plates in triplicate. Corresponding CCR7(+) cells were seeded so that their density was less than 1×10 ⁶ /ml in the case of cells from suspension cultures and 80% confluence was achieved in the case of adherent cells. Cells were harvested (adherent cells were separated with accutase) and resuspended to about 2×10 ⁴ cells/ml. Cells were added to 384-well plates on top of the antibody-payload complex (20 μl/well). The tablet was incubated for four days at 37°C and 5% CO ₂ . A CellTiter-Glo solution (CellTiter-Glo® luminescence cell viability assay; Promega, #G7571) was then prepared and added to the cells at 20 μl/well. Only viable cells produce the ATP required for the luciferase reaction (provided by CellTiter-Glo) that results in luminescence. Accordingly, cell viability was determined from the luminescence signal, which was measured after 10 min incubation at 22°C and 400 rpm using an Envision 2104 multi-label reader. IC50 values were calculated using Graphpad Prism software.

[00456] Figure 7A and Figure 7B show that all four anti-CCR7 antibodies are capable of concentration-dependent cell cytolysis using CCR7+ KE97 cells in a piggyback assay using an MMAF-conjugated reagent. The table below summarizes the results of experiments using MMAF or saporin as a tool-payload in the "piggy-back" format.

Table 16. IC ₅₀ and AMAX of anti-CCR7 antibodies in a cytotoxic assay using pgADC

Antibody to mFc.MMAF Fab-ZAP IC50 (nM) AMAX (%) IC50 (nM) AMAX (%) Original 121G12 0.055 104 0.064 43 Original 506E15 0.070 95 0.135 60 Original 674J13 0.137 102 6.12 31 Original 684E12 0.214 80 2.60 33 MAB197 (R&D) 0.142 85 0.155 40

[00457] The specificity of cytolysis by pgADC was evaluated using target-negative cell lines. Figure 8 shows an example using the FabZap reagent. A specific CCR7-dependent increase in 121G12 pgADC activity was observed in KE97 and NIH3T3.hCCR7 cells, in contrast to CCR7-negative original NIH3T3 cells or control mIgG antibody.

Example 7 Effects of mouse cross-reactive unconjugated or AURIX1-conjugated anti-CCR7 antibodies on normal mouse hematopoietic cells in vivo

[00458] Expression in normal tissues within the species is limited to cells of hematopoietic origin, including CD4+ and CD8+ T cells in the blood and lymphoid organs, making ADC targeting CCR7 potentially unsafe, especially in wild-type Fc format, which could lead to ADCC and depletion of lymphoid cells.

[00459] To determine the effect of targeting ADC to CCR7 found on normal hematopoietic cells in vivo , healthy female CD-1 mice aged 6-8 weeks were evaluated cross-reactive to the mouse of the original Ab 121G12 both in the unconjugated state and conjugated with AURIX1, the Fc format was wild-type or silent (DAPA). Mice were treated with a single IV injection containing wild type 121G12.Cys-Mab.Fc.hIgG1 (121G12.wt.Fc), 121G12.Cys-Mab.DAPA.hIgG1 (121G12.DAPA.Fc) parent, 121G12 parent. Wild-type Cys-Mab.Fc.hIgG1.AURIX1 (121G12.wt.Fc.AURIX1) or original 121G12.Cys-Mab.DAPA.hIgG1.AURIX1 (121G12.DAPA.Fc.AURIX1) at a final dose of 10 mg/ kg. All doses were adjusted according to the individual body weight of the mice.

[00460] On day 25 post-treatment, spleens were removed and separated into single cell suspension using gentleMACS Dissociator (Miltenyi Biotec Inc, San Diego, CA). Then, 1 million cells in each sample were stained with an Ab mixture that contained rat anti-mouse CD8a antibody BUV737, clone 53-6.7 (1:100) (BD Biosciences, San Jose, CA, Cat # 564297) and rat anti-mouse CD4 antibody BV510, clone RM4-5 (1:200) (BD Biosciences, San Jose, CA, Cat # 563106) to determine the effect of individual treatments on CD4+ and CD8a+ T cells. Samples were incubated at 4° C. for 30 min, washed in ice-cold HyClone phosphate buffered saline (Hyclone Laboratories, Logan, UT) and evaluated on a BD LSRFortessa™ cell analyzer (BD Biosciences, San Jose, CA). Total splenocyte counts were used to determine depletion of CD4+ or CD8a+ T cells. T-test was used to determine the significance of differences between groups.

[00461] As shown in Table 17 and Figure 9, a strong reduction in the number of CD4+ (FC 0.5-0.6) and CD8a+ T cells (FC 0.3-0.5) in the spleen was observed on day 3 of treatment with using Ab 121G12.wt.FC or 121G12.wt.Fc.AURIX1 at 10 mg/kg, this suggests that the T cell depletion effect was independent of the presence of the AURIX1 payload. Such effects are avoided by obtaining "silent" Fc by introducing DAPA mutations. Both 121G12.DAPA.Fc and 121G12.DAPA.Fc.AURIX1 had no effect on T cell populations compared to the untreated group. These data indicate that anti-CCR7 ADCs may be unsafe in terms of T-cell depletion, which can be prevented by generating "silent" Fc through DAPA mutations.

Table 17 Effect of 121G12 Antibody on CD4+ and CD8a+ T Cell Populations in CD-1 Mice

The experiment was evaluated on day 25 after treatment. Fold change (FC)=mean splenocyte count on day 25 for indicated treatment group/mean splenocyte count for no-treatment control group on day 3. T-test was used to determine significance versus no treatment group (*p<0.05, ***p<0.001; NS=not significant).

Example 8 ADC activity to CCR7 in direct conjugate format

[00462] Post-binding cytotoxic effects of antibodies (ADC) directly conjugated to various payloads and their internalization into CCR7(+) cells were examined by evaluating cell viability after four days of treatment. Three-fold dilutions of ADC were added to 384-well white bottom plates in triplicate (10 μl/ml). Corresponding CCR7(+) cells were seeded so that their density was less than 1×10 ⁶ /ml in the case of cells from suspension cultures and 80% confluence was achieved in the case of adherent cells. Cells were harvested (adherent cells were separated with accutase) and resuspended to about 2×10 ⁴ cells/ml. Cells were added to 384-well plates on top of the antibody (20 μl/well). The plates were incubated for four days at 37°C and 5% CO ₂ . A CellTiter-Glo solution (CellTiter-Glo® luminescence cell viability assay; Promega, #G7571) was then prepared and added to the cells at 20 μl/well. Cell viability was determined by the luminescence signal, which was measured after 10 min incubation at 22°C and 400 rpm using an Envision reader. IC50 values were calculated using Graphpad Prism software.

[00463] The table below shows examples of cell viability effects measured with CysMab anti-CCR7 Fc wild-type or silent (DAPA) Fc format conjugated to AURIX1 or AURIX2.

Table 18. IC ₅₀ for ADC to CCR7 in a cytotoxic assay

ADC activity in cell viability assay; IC ₅₀ (nM) cell line type of cancer 506E15.CysMab.AURIX1 674J13.
CysMab. AURIX1 506E15.
CysMab.DAPA.
AURIX2 674J13.
CysMab.DAPA.AURIX2 DEL ALCL 0.0029 0.0741 0.0084 0.1167 KE97 multiple myeloma 0.0031 0.03 0.0131 0.0658

[00464] To evaluate ADC for target specificity and dependence on receptor level, ADC activity was tested on cell lines with different levels of CCR7 receptors. The table below shows an example of a humanized 674J13 antibody in CysMab.DAPA format and conjugated to AURIX2. Cell lines were selected based on similar payload sensitivity.

Table 19. IC _{50 of} humanized 674J13 antibody in DAPA format and conjugated to AURIX2

ADC activity in cell viability assay; IC50 (nM) cell line type of cancer Levels of CCR7 receptors 674J13.CysMab.DAPA.AURIX2 DEL ALCL ~100,000 0.2417 (>95% of AMAX) KE97 multiple myeloma ~100,000 0.1018 (>95% of AMAX) SR786 Anaplastic large cell T-cell lymphoma ~28,000 2,737 (90% of AMAX) CML-T1 T-cell leukemia ~29,000 3,779 (60% of AMAX) DND-41 T-cell leukemia 1,700 < 20% off AMAX NCI-H82 Small cell lung cancer 0 Absence of cytolysis

[00465] As seen from the pHrodo experiment, ADC activity requires a higher degree of receptor number than the ADCC mechanism. The exact cut-off for the number of receptors depends on various parameters (eg, antibody avidity, payload activity), and the data shown in this example describes general principles. The use of an avidity-dependent antibody to CCR7 in DAPA format shifts ADC activity towards cancer cells rather than normal CCR7+ PBMCs, which in this example are cancer cell lines with less than 2000 CCR7 receptors.

Example 9 Site Specific Introduction of MPET.DM4 into ADC

[00466] DM4 conjugated ADCs are conventional in the ADC art. In this example, the present inventors describe the construction and use of site-specifically conjugated MPET.DM4 using a CysMab variant antibody, which provides the advantage of obtaining a reproducibly homogeneous lot of ADC with controlled DAR, in which the DAR (drug to antibody ratio) is approximately 4. It was described that conjugates prepared without the use of site-specific conjugation often contain significant high DAR populations that have been associated with undesirable biophysical features, including increased hydrophobicity and hence faster clearance, an unsatisfactory PK profile, and increased toxicity. Below, the authors of the present invention show different in vitro and in vivo estimates of ADC to CCR7 based on MPET.DM4 in comparison with its counterpart based on sSPDB.DM4.

Example 10 Binding Affinity of ADC to MPET.DM4 Versus ADC to sSPDB.DM4 in a FACS Analysis

[00467] Another potential beneficial effect of site-specific conjugation versus no site-specific conjugation could be the potential interference of lysine conjugation sites that are structurally located near important CSD residues. Payload conjugation to such lysine sites would be expected to affect ADC binding affinity. To test the binding affinities of ADCs using MPET.DM4-conjugated CysMabs described in this example versus antibodies conjugated to sSPDB.DM4 via endogenous lysine, binding affinities were determined by FACS as described above. The table below summarizes some exemplary affinity data that shows a modest reduction in binding affinity for sSPDB.DM4 ADC compared to MPET.DM4 ADC.

Table 20 ADC binding affinity for CCR7

Affinity of ADC in FACS analysis; EC50 (nM) cell line Levels of CCR7 receptors 121G12.
CysMab.DAR4, unconjugated 121G12.
CysMab.DAPA.
MPET.DM4
(DAR 3.8) 121G12.
DAPA.sSPDB.DM4 (DAR3,9) DEL ≥100,000 5.24 5.61 8.42

Example 11. In vitro ADC activity with MPET.DM4 compared to ADC with sSPDB.DM4

[00468] The cytotoxic effects of 121G12.CysMab.DAPA.MPET.DM4 and 121G12.DAPA.sSPDB.DM4 (DAR3.9) ADCs were assessed in an in vitro viability assay as described above. The table below shows a similar, slightly improved activity of the MPET.DM4 conjugate compared to the sSPDB.DM4 conjugate. This may be due to better preserved affinity or other factors.

Table 21. In vitro cytotoxic activity of ADC to CCR7

ADC activity in cell viability assay; IC50 (nM) cell line 121G12.CysMab.DAPA.MPET.DM4 121G12.DAPA.sSPDB.DM4 (DAR3.9) L540 2.944 3.905 DEL 2.286 2.983 KE97 2.171 3.177

[00469] ADC 121G12.CysMab.DAPA.MPET.DM4 was further tested on a number of cancer cell lines covering various indications where it achieved significant cell cytolysis that correlated with receptor density.

Table 22. In vitro cytotoxic activity of ADC to CCR7 in cell lines

cell line type of cancer Relative density of receptors (%) IC50 121G12.CysMab.DAPA.MPET.DM4 (nM) SUPHD1 Hodgkin's lymphoma 310 1.69 L540 Hodgkin's lymphoma 175 4.90 KE97 multiple myeloma 100 2.24 JVM2 MCL 79 4.62 MOTN1 CLL 67 4.24 DEL ALCL 67 2.28 OCI-Ly3 ABC-DLBCL 43 6.42 Toledo DLBCL twenty n.d. Mec-2 CLL fifteen >20 PEER T-ALL 2 >20

Example 12 Dose Dependent in vivo efficacy of 121G12.CysMab.DAPA.MPET.DM4 and 121G12.DAPA.sSPDB.DM4 against the KE97 xenograft model of multiple myeloma in SCID-Beige mice

[00470] To demonstrate the in vivo targeted antitumor activity of 121G12.CysMab.DAPA.MPET.DM4 and 121G12.DAPA.sSPDB.DM4, a KE97 xenograft model was engrafted to female SCID-beige mice by subcutaneous injection of 3×10 ⁶ cells into the right flank of each mouse . After tumors reached approximately 135 mm ³ , mice were randomized according to tumor volume into treatment groups (n=8 per group). Mice were treated with IV administration of one of 121G12.CysMab.DAPA.MPET.DM4 (DAR4) at a final dose of 0.5, 2 or 5 mg/kg, 121G12.DAPA.sSPDB.DM4 (DAR 3.9) based 0.5, 2 or 5 mg/kg or non-specific isotype control IgG1.CysMab.DAPA.MPET.DM4 at 5 mg/kg. All doses were adjusted according to the individual body weight of the mice.

[00471] All tested agents were well tolerated throughout the study, and no significant clinical symptoms of toxicity or weight loss were observed in any of the treatment groups (Table 23).

Table 23. Dose-dependent Efficacy of Anti-CCR7 ADCs in the KE97 Xenograft Model

Tumor reaction Host reaction treatment Dose, scheme of administration ΔT/ΔC (%) Regression (%) Δ body weight (%) Survival (live/total if
quality) No processing Missing 100 - 2.09 8/8 IgG1.CysMab.DAPA.MPET.DM4 5 mg/kg
single dose 92.94 - 3.02 8/8 121G12.CysMab.DAPA.MPET.DM4 0.5 mg/kg
single dose 78.62 - 1.05 8/8 121G12.CysMab.DAPA.MPET.DM4 2 mg/kg
single dose - 33.07* 2.74 8/8 121G12.CysMab.DAPA.MPET.DM4 5 mg/kg
single dose - 53.66* 0.46 8/8 121G12.DAPA.sSPDB.DM4 0.5 mg/kg
single dose 85.38 - 1.84 8/8 121G12.DAPA.sSPDB.DM4 2 mg/kg
single dose 13.77* - 0.59 8/8 121G12.DAPA.sSPDB.DM4 5 mg/kg
single dose - 32.63* 3.63 8/8

The experiment was evaluated at day 9 post-treatment (day 23 post-implantation), *p<0.001 compared to no-treatment control (one-way ANOVA/Tukey's multiple comparison test). % ΔT/ΔC=100 ΔT/ΔC where: ΔT=mean tumor volume in the drug-treated group in study D23 - the mean tumor volume in the drug-treated group on the initial day of dosing; ΔC=mean tumor volume in the control group in the D23 study - the average tumor volume in the control group on the initial day of dosing D14. % regression=(1-T final/T initial)×100, where T final is the mean tumor volume at D23 and T initial is defined as the tumor volume at D14 after implantation. Δ body weight (%)=(average body weight in D23-average body weight in D14) *100/average body weight in D14 treatments.

[00472] Negligible antitumor efficacy was observed after treatment with non-specific isotype control IgG1.CysMab.DAPA.MPET.DM4 at 5 mg/kg. Treatment with 121G12.CysMab.DAPA.MPET.DM4 resulted in a dose-dependent antitumor efficacy with a ΔT/ΔC value of 78.62% (0.5 mg/kg), while doses of 2 and 5 mg/kg resulted in a mean regression by 33% and 54%, respectively, to D9 after the first dose (D23 after implantation). Treatment with 121G12.DAPA.sSPDB.DM4 also demonstrated dose-dependent antitumor efficacy with ΔT/ΔC values of 85.38% (0.5mg/kg) and 13.77% (2mg/kg), while the dose was 5 mg/kg resulted in a mean regression of 33% to D9 after the first dose (D23 after implantation). Ko D23-D25 after implantation mice in the control and 0.5 mg/kg treatment groups were sacrificed and the remaining groups were treated with a second dose in D28 after implantation of one of 121G12.CysMab.DAPA.MPET.DM4 at 2 or 5 mg/kg or 121G12.DAPA.sSPDB.DM4 at the rate of 2 or 5 mg/kg. Sustained tumor regression was observed in the case of administration of 121G12.CysMab.DAPA.MPET.DM4 at 2 and 5 mg/kg, as well as in the case of 121G12.DAPA.sSPDB.DM4 at 5 mg/kg until the end of the study in D42 after implantation . In the case of 121G12.DAPA.sSPDB.DM4 at 2 mg/kg, the response was more heterogeneous, with approximately 25% of the mice showing stable tumor regression, while the rest of the group showed either disease stabilization or tumor progression (Figure 10, table 23).

Example 13 Efficacy Evaluation of 121G12.CysMab.DAPA.MPET.DM4 Versus 121G12.DAPA.sSPDB.DM4 in the KE97 Multiple Myeloma Model at Large Initial Tumor Volumes

[00473] A more demanding in vivo model was provided using the KE97 multiple myeloma cell line to further differentiate antitumor efficacy due to different cleavable linkers by comparing the potency of 121G12.CysMab.DAPA.MPET.DM4 to 121G12.DAPA.sSPDB. DM4 (DAR3,9) against tumors with large initial tumor volumes at the first dose. The KE97 xenograft model was engrafted to female SCID-beige mice by subcutaneous injection of 3×10 ⁶ cells into the right flank of each mouse. After tumors reached approximately 450 mm ³ , mice were randomized according to tumor volume into two treatment groups (n=8). Mice were treated with IV administration of 2 mg/kg of either 121G12.CysMab.DAPA.MPET.DM4 or 121G12.DAPA.sSPDB.DM4.

[00474] Negligible antitumor efficacy was observed after treatment with 121G12.DAPA.sSPDB.DM4 at 2 mg/kg. Treatment with 121G12.CysMab.DAPA.MPET.DM4 resulted in partial regression/prolonged stunting in 75% (6 out of 8) single dose treated mice (Figure 11). 121G12.CysMab.DAPA.MPET.DM4 strongly outnumbered 121G12.DAPA.sSPDB.DM4 in this model (in D25, the mean tumor volume was 524.83 ± 143.20 vs. 1337.13 ± 35.13, respectively, p< 0.001; unpaired T-test).

Example 14 in vivo efficacy of 121G12.CysMab.DAPA.MPET.DM4 against a tumor model based on patient-derived primary non-small cell lung cancer HLUX1934

[00475] The antitumor activity of 121G12.CysMab.DAPA.MPET.DM4 was evaluated in a xenograft model of primary non-small cell lung cancer HLUX1934 expressing CCR7. Female athymic nude mice were implanted subcutaneously with tumor fragments in the right flank of each mouse. After tumors reached approximately 100 mm ³ , mice were randomized according to tumor volume into treatment groups (n=8). Mice were treated with IV administration of one of the 121G12.CysMab.DAPA.MPET.DM4 (DAR4) at 10 mg/kg or the non-specific isotype control IgG1.CysMab.DAPA.MPET.DM4 at 10 mg/kg. A second dose of each antibody was administered 2 weeks later. All doses were adjusted according to the individual body weight of the mice.

[00476] The non-specific isotype control IgG1.CysMab.DAPA.MPET.DM4 at 10 mg/kg appeared to result in a slight delay in tumor growth compared to the no-treatment group (ΔT/ΔC value was 53.07%), potentially due to non-specific binding of the antibody to a non-target target in the HLUX1934 model. Treatment with 121G12.CysMab.DAPA.MPET.DM4 resulted in a more pronounced efficacy, which was sustained at the second dose. Treatment with 121G12.CysMab.DAPA.MPET.DM4 at 10mg/kg was well tolerated with no apparent weight loss and a ΔT/ΔC value of 18.83% in D34 after implantation (Figure 12, Table 24).

Table 24 Efficacy of 121G12.CysMab.DAPA.MPET.DM4 against patient-derived NSCLC model HLUX1934. The experiment was evaluated at day 34 post-implantation (D23 post-treatment)

Tumor reaction Host reaction Treatment Dose ΔT/ΔC (%) Δ body weight (%) Survival (live/total) No processing Missing 100 5.12 5/8** IgG1.CysMab.DAPA.MPET.DM4 10 mg/kg 53.07 2.81 8/8 121G12.CysMab.DAPA.MPET.DM4 10 mg/kg 18.83* 0.61 8/8

* p<0.001 compared to untreated controls (one-way ANOVA/Tukey's multiple comparison test). % ΔT/ΔC=100 ΔT/ΔC where: ΔT=mean tumor volume in the drug-treated group in study D34 - mean tumor volume in the drug-treated group on the initial day of dosing; ΔC=mean tumor volume in the control group in the D34 study - the average tumor volume in the control group on the initial day of dosing. Δ body weight (%)=(average body weight in D34-average body weight in D11) *100/average body weight in D11 treatment. ** In the no-treatment group, mice were sacrificed due to excess tumor mass in D25-D27 after treatment.

Example 15 in vivo efficacy of 684E12.SMCC.DM1 against the KE97 xenograft model of multiple myeloma in SCID-Beige mice

[00477] To demonstrate the in vivo targeted antitumor activity of 684E12.SMCC.DM1, a KE97 xenograft model was engrafted to female SCID-beige mice by subcutaneous injection of 3×10 ⁶ cells into the right flank of each mouse. After tumors reached approximately 200 mm ³ , mice were randomized according to tumor volume into treatment groups (n=8 per group). Mice were treated with IV administration of one of the parent 684E12.SMCC.DM1 (DAR2,6) at a final dose of 2 or 6 mg/kg or the non-specific isotype control IgG1.SMCC.DM1 at 6 mg/kg. All doses were adjusted according to the individual body weight of the mice.

[00478] All tested agents were well tolerated throughout the study, and no significant clinical symptoms of toxicity or weight loss were observed in any of the treatment groups. Negligible antitumor efficacy was observed after treatment with non-specific isotype control IgG1.SMCC.DM1 or parent 684E12.SMCC.DM1 at 2 mg/kg. Treatment with stock 684E12.SMCC.DM1 at 6 mg/kg resulted in a ΔT/ΔC value of 6.72% in D11 after dosing (p<0.0001, one-way ANOVA/Tukey's test for multiple comparisons) (Figure 13) .

Example 16 in vivo Targeted modulation of pharmacodynamic markers by 121G12.CysMab.DAPA.MPET.DM4 in the KE97 tumor model

[00479] Accumulation of phospho-histone H3 positive tumor cells after treatment with 121G12.CysMab.DAPA.MPET.DM4 was used to evaluate the ability of CCR7 ADCs to induce G2/M arrest in vivo .

[00480] A study was conducted in which a KE97 xenograft model was engrafted to female SCID-beige mice by subcutaneous injection of 3×10 ⁶ cells into the right flank of each mouse. After tumors reached approximately 140 mm ³ , mice were randomized according to tumor volume to treatment groups (n=3 per group). Mice were treated with a single IV injection of one of the 121G12.CysMab.DAPA.MPET.DM4 at a final dose of 2, 5 or 10 mg/kg or the non-specific isotype control IgG1.CysMab.DAPA.MPET.DM4 at 10 mg/kg. All doses were adjusted according to the individual body weight of the mice. 48 hours post-treatment, tumors were harvested for phospho-histone H3 levels by immunohistochemical staining, described below.

[00481] In order to measure the accumulation of phospho-histone H3 positive nuclei by immunohistochemistry, a rabbit polyclonal antibody targeting residues surrounding human histone H3 phosphorylated serine 10 was obtained from Ventana Medical Systems (Tucson, Arizona, cat no. 760-4591). The IHC protocol involved heating and moderate exposure (32 min) to Ventana Discovery Cell Conditioner 1 antigen unmasking reagent. Samples were incubated for 60 min at room temperature with primary antibody (pre-diluted according to manufacturer's recommendations). A subsequent 12 min incubation was performed with HRP-conjugated OmniMap rabbit secondary Ab (Ventana, Tucson, AZ, Cat # 760-4311) (pre-diluted according to manufacturer's recommendations).

[00482] Although Figure 14A shows rare phospho-histone H3 positive tumor cells in preparations from the no-treatment group and the IgG1.CysMab.DAPA.MPET.DM4 isotype control group, a consistent dose-dependent increase in phospho-histone immune staining H3 was detected 48 hours after the introduction of 121G12.CysMab.DAPA.MPET.DM4. Signal quantification was performed using MatLab (MathWorks, Natick, Massachusetts) where total phospho-histone H3 signal area (µm ² ) was normalized to total nuclear area (µm ² ) to give relative phospho-histone H3 values (%), shown below for each of the treatment groups. The data in Figure 14B indicate that 121G12.CysMab.DAPA.MPET.DM4 is able to induce a pronounced arrest at the G2/M stage on tumor xenografts, consistent with the expected payload mechanism.

Example 17. Method for obtaining antibodies 121G12.CysMab.DAPA

[00483] This example describes a method for producing an anti-CCR7 antibody, 121G12.CysMab.DAPA, using cell culture, where Ab is expressed from a vector that encodes Ab. After Ab was expressed in cell culture, Ab was purified from cell culture as follows.

[00484] The first step of the method for purifying the 121G12.CysMab.DAPA drug substance intermediate was to remove the cells by in-line deep filtration, followed by filtration through a 0.2 µm filter.

[00485] The second step is the Protein A affinity liquid chromatography step. Depending on the total amount of bulk product, this step is carried out in several cycles. Each cycle allows for a maximum loading of approximately 20 g/L column volume. Elution was carried out with 50 mM acetic acid, approximately pH 3.0. The operating temperature is 18-28°C. All eluates are pooled and stored at 2-8° C. until the virus inactivation step is carried out.

[00486] The third step is virus inactivation by "low pH treatment". The temperature of the intermediate solution from step 2 is brought to 18-28° C. and the pH is adjusted to 3.5 (range 3.4-3.6). The product intermediate solution is then allowed to inactivate the virus for 70 minutes (range 60-90 minutes). After the end of the aging time, the solution is adjusted to pH 6.0 (range 5.8-6.2). At the end of the stage, the solution is subjected to deep flow filtration using a filter with a pore size of 0.2 μm and stored at 2-8°C.

[00487] The fourth step is cation exchange chromatography in the binding/elution mode, which provides for integrated recovery on the column. Depending on the titer, this step is carried out in several cycles. Each cycle allows for a loading of approximately 30 g/L column volume. The column is equilibrated with buffer A containing 20 mM sodium succinate, pH 6.0. Recovery on the column is carried out using 20 mm sodium phosphate, 1 mm EDTA, 7 mm L-cysteine, pH 7.1, as a recovery buffer. The recovery buffer is removed with buffer A and the elution is carried out with a linear gradient from 10% to 90% with buffer A and buffer B containing 10 mm sodium succinate, 300 mm sodium chloride, pH 6.0. The eluates and pooled eluates can be stored at 2-8° C. until the anion exchange chromatography step on multimodal separation matrices.

[00488] The fifth step of the method is "continuous flow" anion exchange chromatography. Depending on the titer, this step is carried out in several cycles. Each cycle allows for a maximum load of approximately 350 g/L column volume. The operating temperature is 18-28°C. Equilibration is carried out with 20 mM sodium succinate, 119 mM sodium chloride, pH 6.0. The final filtration product is stored at 2-8° C. prior to the virus removal step.

[00489] Virus filtration, the sixth stage, consists of prefiltration with a filter with a pore size of 0.1 μm, followed by virus filtration with a Planova 20N nanofilter. The temperature of the intermediate solution from step 5 is adjusted to 18-28° C. before viral filtration. The operating temperature is 18-28°C. After nanofiltration, the intermediate is stored at 2-8°C or 18-28°C.

[00490] The seventh step, ultrafiltration/diafiltration, consists of a concentration step to about 70 g/L followed by a ^1st diafiltration step with 10 mM potassium phosphate, pH 6.0. With diafiltration, it is planned to obtain an exchange ratio of at least 7, followed by dilution to about 50 g/l. The final drug substance intermediate is passed through a 0.2 µm filter and stored at 2-8°C.

[00491] In the eighth and final step, the final bulk drug substance intermediate is aliquoted into suitable containers and stored below -60°C after freezing.

Table 25. Flowchart of the cleaning and recovery method

Stage Operation Stage 1 Collection, removal of cells and filtration Stage 2 Affinity chromatography
(MabSelect SuRE) Stage 3 Virus inactivation at pH 3.5 Stage 4 Cation exchange chromatography
and
column recovery
(Fractogel EMD SO3(M)) Stage 5 Anion exchange chromatography on multimodal separation matrices
(Capto adhere) Stage 6 Virus removal by nanofiltration
(Planova 20N) Stage 7 Ultrafiltration/diafiltration and final filtration Stage 8 Filling and deep freezing

Example 18 In vivo dose dependent efficacy of 121G12.CysMab.DAPA.MPET.DM4 against the ABC-DLBCL OCI-LY3 xenograft model in NSG mice.

[00492] To demonstrate the in vivo targeted antitumor activity of 121G12.CysMab.DAPA.MPET.DM4 in the ABC-DLBCL model, the OCI-LY3 xenograft model was engrafted to female NSG mice by subcutaneous injection of 10×10 ⁶ cells into the right flank of each mouse. After tumors reached approximately 140 mm ³ , mice were randomized according to tumor volume into treatment groups (n=6 per group). Mice were treated with IV administration of one of the 121G12.CysMab.DAPA.MPET.DM4 (DAR4) at a final dose of 0.5, 1, or 2 mg/kg or the non-specific isotype control hIgG1.CysMab.DAPA.MPET.DM4 at 2 mg/kg. kg on day 1 and day 15 of the study. All doses were adjusted according to the individual body weight of the mice. All tested agents were well tolerated throughout the study, and no significant clinical signs of toxicity or weight loss were observed in any of the treatment groups (Table 26).

[00493] Negligible antitumor efficacy was observed after treatment with the non-specific isotype control hIgG1.CysMab.DAPA.MPET.DM4 at 2 mg/kg. Treatment with 121G12.CysMab.DAPA.MPET.DM4 resulted in dose-dependent antitumor efficacy with ΔT/ΔC values of 74.6% (0.5 mg/kg) and 10.7% (1 mg/kg), while the 2 mg/kg dose resulted in a mean regression of 65.9% by day 28 of the study. 3 out of 6 mice in the 121G12.CysMab.DAPA.MPET.DM4 group at 2 mg/kg showed complete regression (FIG. 15).

Table 26. Dose-dependent efficacy of anti-CCR7 ADC against the OCI-LY3 xenograft model at day 28 of treatment.

Tumor response Host reaction Treatment Dose, scheme of administration ∆T/∆C (%) Regression (%) Δ body weight (%) Survival (live/
total) No processing Missing 100.0 - 5.6 6/6 hIgG1.CysMab.DAPA.MPET.DM4 2 mg/kg
when dosing at D1 and D15 119.7 - 1.8 5/6 121G12.CysMab.DAPA.MPET.DM4 0.5 mg/kg
when dosing at D1 and D15 74.6 - 3.5 5/6 121G12.CysMab.DAPA.MPET.DM4 1 mg/kg
when dosing at D1 and D15 10.7** 2.9 6/6 121G12.CysMab.DAPA.MPET.DM4 2 mg/kg
when dosing at D1 and D15 - 65.9** -0.8 6/6

The experiment was evaluated on day 28 post-treatment, **p<0.005 compared to no-treatment control (one-way ANOVA/Tukey's multiple comparison test). % ΔT/ΔC=100 ΔT/ΔC where: ΔT=mean tumor volume in the drug-treated group in study D28 - the mean tumor volume in the drug-treated group on the initial day of dosing; ΔC=mean tumor volume in the control group in the D28 study - the average tumor volume in the control group on the initial day of dosing D1. % regression=(1-T _final /T _initial ) x100 was calculated if ΔT<0, where T _final is the mean tumor volume in D28 and T _initial is defined as the tumor volume in D1 treatment. Δ body weight (%)=(average body weight in D28-average body weight in D1) *100/average body weight in D1 treatment.

Example 19 Dose-dependent in vivo efficacy of 121G12.CysMab.DAPA.MPET.DM4 against the GCB-DLBCL Toledo xenograft model in SCID-bg mice.

[00494] To demonstrate the in vivo targeted antitumor activity of 121G12.CysMab.DAPA.MPET.DM4 in the GCB-DLBCL model, the Toledo xenograft model was engrafted to female Scid-bg mice by subcutaneous injection of 3×10 ⁶ cells into the right flank of each mouse. After tumors reached approximately 100 mm ³ , mice were randomized according to tumor volume to treatment groups (n=4 per group). Mice were treated with IV administration of one of the 121G12.CysMab.DAPA.MPET.DM4 (DAR4) at a final dose of 2 or 5 mg/kg or the non-specific isotype control hIgG1.CysMab.DAPA.MPET.DM4 at 5 mg/kg on day 1 and day 15 of the study. All doses were adjusted according to the individual body weight of the mice. All tested agents were well tolerated throughout the study, and no significant clinical signs of toxicity or weight loss were observed in any of the treatment groups (Table 27).

[00495] Negligible antitumor efficacy was observed after treatment with the non-specific isotype control hIgG1.CysMab.DAPA.MPET.DM4 at 5 mg/kg. Treatment with 121G12.CysMab.DAPA.MPET.DM4 resulted in dose-dependent antitumor efficacy with ΔT/ΔC values of 52.7% (2 mg/kg) and 20.6% (5 mg/kg) (Figure 16, Table 27) .

Table 27. Dose-dependent efficacy of anti-CCR7 ADC against the Toledo xenograft model at day 20 of treatment.

Tumor response Host reaction Treatment Dose, scheme of administration ΔT/ΔC (%) Δ body weight (%) Survival (live/total) No processing Missing 100.0 4.9 3/6 hIgG1.CysMab.DAPA.MPET.DM4 5 mg/kg
when dosing at D1 and D15 94.5 1.0 3/6 121G12.CysMab.DAPA.MPET.DM4 2 mg/kg
when dosing at D1 and D15 52.7* 3.2 4/6 121G12.CysMab.DAPA.MPET.DM4 5 mg/kg
when dosing at D1 and D15 20.6** 0.1 4/6

The experiment was evaluated at day 20 post-treatment, *p<0.05, **p<0.005 compared to no-treatment controls (one-way ANOVA/Tukey's multiple comparison test). % ΔT/ΔC=100 ΔT/ΔC where: ΔT=mean tumor volume in the drug-treated group in study D20 - the mean tumor volume in the drug-treated group on the initial day of dosing; ΔC=mean tumor volume in the control group in the D20 study - the average tumor volume in the control group on the initial day of dosing D1. Δ body weight (%)=(average body weight in D20-average body weight in D1) *100/average body weight in D1 treatment.

Example 20 In vivo efficacy of 121G12.CysMab.DAPA.MPET.DM4 in a DEL ALCL xenograft model in SCID-bg mice.

[00496] To demonstrate the in vivo targeted antitumor activity of 121G12.CysMab.DAPA.MPET.DM4 in a CCR7 positive ALCL model, a DEL xenograft model was engrafted to female Scid-bg mice by subcutaneous injection of 3x10 ⁶ cells into the right flank of each mouse. After tumors reached approximately 100 mm ³ , mice were randomized according to tumor volume to treatment groups (n=4 per group). Mice were treated with IV administration of one of the 121G12.CysMab.DAPA.MPET.DM4 (DAR4) at a final dose of 2 mg/kg or the non-specific isotype control isotype.MPET.DM4 at 2 mg/kg on day 1 and day 15 of the study. All doses were adjusted according to the individual body weight of the mice.

[00497] Negligible antitumor efficacy was observed after treatment with the non-specific isotype control hIgG1.CysMab.DAPA.MPET.DM4 at 2 mg/kg. Treatment with 121G12.CysMab.DAPA.MPET.DM4 at 2 mg/kg resulted in a mean regression of 40.2% by study day 14 after a single dose (p<0.01). Mice received a second dose on day 15 and were observed for an additional three weeks. One exception animal did not respond to the second dose treatment and had to be sacrificed on day 28 due to tumor mass. Another animal showed slow disease progression and was also excluded on day 28 from follow-up for target expression. Two of the four mice continued to observe a sustained effect on tumor growth (FIG. 17). All treatments were well tolerated with no apparent weight loss.

Example 21 In Vivo Efficacy of 121G12.CysMab.DAPA.MPET.DM4 in a Tumor Model Based on Patient-Derived Primary Non-Small Cell Lung Cancer HLUX1787

[00498] The antitumor activity of 121G12.CysMab.DAPA.MPET.DM4 was evaluated in a xenograft model of primary non-small cell lung cancer HLUX1787 expressing CCR7. Female NSG mice were implanted subcutaneously with tumor fragments in the right flank of each mouse. After tumors reached approximately 150 mm ³ , mice were randomized according to tumor volume into treatment groups (n=6 per group). Mice were treated with IV administration of one of 121G12.CysMab.DAPA.MPET.DM4 (DAR4) at 0.5, 2, or 5 mg/kg on day 1, and a second dose was given 2 weeks later on day 15. All doses were adjusted according to with individual body weight of mice.

[00499] Insignificant antitumor efficacy was observed with low doses of conjugated Ab, however, partial regression or stabilization of the disease was observed with treatment with 121G12.CysMab.DAPA.MPET.DM4 at 5 mg/kg. Sustained tumor efficacy was observed two weeks after the second dose, resulting in a ΔT/ΔC value of 1.3% in treatment D30 (p<0.001; 1-way ANOVA/Tukey's multiple comparison test) (Figure 18). All treatments were well tolerated with no apparent weight loss.

--->

SEQUENCE LIST

<110> NOVARTIS AG

<120> ANTIBODY TO CCR7 AND DRUG CONJUGATES

<130> PAT057594-WO-PCT

<140>

<141>

<150> 62/454.476

<151> 2017-02-03

<160> 629

<170> PatentIn version 3.5

<210> 1

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 1

Gly Phe Thr Phe Ser Tyr Ala Met Ser

1.5 10

<210> 2

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 2

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 3

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 3

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 4

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 4

Ser Tyr Ala Met Ser

fifteen

<210> 5

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 5

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 6

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 6

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 7

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 7

Gly Phe Thr Phe Ser Ser Tyr

fifteen

<210> 8

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 8

Ser Ser Gly Gly Ser Phe

fifteen

<210> 9

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 9

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 10

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 10

Gly Phe Thr Phe Ser Ser Tyr Ala

fifteen

<210> 11

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 11

Ile Ser Ser Gly Gly Ser Phe Thr

fifteen

<210> 12

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 12

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 13

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 13

Glu Val Gln Leu Val Glu 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 Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 14

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 14

gaagtgcagc tggtggaatc tggcggcgga ctggtcaagc ctggcggctc cctgagactg 60

tcttgcgccg cctccggctt caccttctcc agctacgcca tgtcctggat ccggcaggcc 120

cctggcaagg gactggagtg ggtggccacc atctcctccg gcggcagctt cacctactac 180

cccgactccg tgaagggccg gttcaccatc tcccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacgggcc 300

tccaccgtcg tgggcaccga tttcgatgtg tggggccagg gcacaaccgt gaccgtgtcc 360

tcc 363

<210> 15

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 15

Glu Val Gln Leu Val Glu 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 Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 16

<211> 1353

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 16

gaagtgcagc tggtggaatc tggcggcgga ctggtcaagc ctggcggctc cctgagactg 60

tcttgcgccg cctccggctt caccttctcc agctacgcca tgtcctggat ccggcaggcc 120

cctggcaagg gactggagtg ggtggccacc atctcctccg gcggcagctt cacctactac 180

cccgactccg tgaagggccg gttcaccatc tcccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacgggcc 300

tccaccgtcg tgggcaccga tttcgatgtg tggggccagg gcacaaccgt gaccgtgtcc 360

tccgcctcca ccaagggacc ctccgtgttc cctctggccc cttccagcaa gtccacctct 420

ggcggcaccg ccgctctgggg ctgcctggtc aaggactact tcccctgccc tgtgacagtg 480

tcctggaact ccggcgctct gacctccggc gtgcacacct tccctgccgt gctgcagtcc 540

tccggcctgt actccctgtc ctccgtcgtg accgtgcctt cctccagcct gggcacccag 600

acctacatct gcaacgtgaa ccacaagccc tccaacacca aagtggacaa gcgggtggaa 660

cccaagtcct gcgacaagac ccacacctgt cctccctgcc ctgcccctga gctgctggga 720

ggcccttccg tgttcctgtt ccctccaaag cccaaggaca ccctgatgat ctcccggacc 780

cctgaagtga cctgcgtggt ggtggccgtg tcccacgagg atcccgaagt gaagttcaat 840

tggtacgtgg acggcgtgga agtgcacaat gccaagacca agcccagaga ggaacagtac 900

aactccacct accgggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 960

aaagagtaca agtgcaaagt gtccaacaag gccctggccg ctcccatcga aaagaccatc 1020

tccaaggcca agggccagcc cagagagccc caagtgtaca cactgcctcc cagccgggaa 1080

gagatgacca agaaccaagt gtccctgacc tgcctcgtga agggcttcta cccctgcgat 1140

atcgccgtgg agtgggagtc caacggccag cccgagaaca actacaagac cacccctccc 1200

gtgctggaca gcgacggctc attcttcctg tactccaagc tgaccgtgga caagtcccgg 1260

tggcagcagg gcaacgtgtt ctcctgctcc gtgatgcacg aggccctgca caaccactac 1320

acccagaagt ccctgtccct gagccccggc aag 1353

<210> 17

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 17

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 18

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 18

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 19

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 19

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 20

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 20

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 21

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 21

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 22

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 22

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 23

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 23

Ser Gln Asp Ile Gly Ser Ser

fifteen

<210> 24

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 24

Ala Thr Ser

one

<210> 25

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 25

Tyr Ala Ser Ser Pro

fifteen

<210> 26

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 26

Gln Asp Ile Gly Ser Ser

fifteen

<210> 27

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 27

Ala Thr Ser

one

<210> 28

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 28

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 29

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 29

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1.5 10 15

Asp Arg Val Thr Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Lys Pro Gly Lys Ala Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 30

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 30

gacatccaga tgacccagag cccctccagc ctgtccgcct ccgtgggcga tagagtgacc 60

ctgacctgcc gggcctccca ggacatcggc tcctccctga actggctgca gcagaagccc 120

ggcaaggcca tcaagcggct gatctacgcc acctcctccc tggactccgg cgtgccctcc 180

cggttctctg gctccagatc cggcaccgac tacaccctga ccatctccag cctgcagccc 240

gaggacttcg tggtgtacta ctgcctgcag tacgcctcca gccctcccac cttcggcgga 300

ggcaccaagc tggaaatcaa g 321

<210> 31

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 31

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1.5 10 15

Asp Arg Val Thr Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Lys Pro Gly Lys Ala Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 32

<211> 642

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 32

gacatccaga tgacccagag cccctccagc ctgtccgcct ccgtgggcga tagagtgacc 60

ctgacctgcc gggcctccca ggacatcggc tcctccctga actggctgca gcagaagccc 120

ggcaaggcca tcaagcggct gatctacgcc acctcctccc tggactccgg cgtgccctcc 180

cggttctctg gctccagatc cggcaccgac tacaccctga ccatctccag cctgcagccc 240

gaggacttcg tggtgtacta ctgcctgcag tacgcctcca gccctcccac cttcggcgga 300

ggcaccaagc tggaaatcaa gcgtacggtg gccgctccca gcgtgttcat cttccccccc 360

agcgacgagc agctgaagag cggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420

ccccgggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480

gagagcgtca ccgagcagga cagcaaggac tccacctaca gcctgagcag caccctgacc 540

ctgagcaagg ccgactacga gaagcataag gtgtacgcct gcgaggtgac ccaccagggc 600

ctgtccagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642

<210> 33

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 33

Gly Phe Thr Phe Ser Thr Tyr Ala Met Ser

1.5 10

<210> 34

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 34

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 35

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 35

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 36

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 36

Thr Tyr Ala Met Ser

fifteen

<210> 37

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 37

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 38

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 38

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 39

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 39

Gly Phe Thr Phe Ser Thr Tyr

fifteen

<210> 40

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 40

Ser Asp Ala Gly Ser Tyr

fifteen

<210> 41

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 41

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 42

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 42

Gly Phe Thr Phe Ser Thr Tyr Ala

fifteen

<210> 43

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 43

Ile Ser Asp Ala Gly Ser Tyr Ser

fifteen

<210> 44

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 44

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10 15

<210> 45

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 45

Glu Val Gln Leu Val Glu 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 Phe 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 Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Glyn Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 46

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 46

gaagtgcagc tggtggaatc tggcggcgga ctggtcaagc ctggcggctc cctgagactg 60

tcttgcgccg cctccggctt caccttctcc acctacgcca tgtcctggggt ccgacaggcc 120

cctggaaagg gcctggagtg ggtggccacc atctccgacg ccggctccta ctcctactac 180

cccgacaacg tgaagggccg gttcaccatc tcccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgcgc cagacggggc 300

tccagatacg aagagtacta cgtgatggac tactggggcc agggcacaac cgtgaccgtg 360

tcctcc 366

<210> 47

<211> 452

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 47

Glu Val Gln Leu Val Glu 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 Phe 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 Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 48

<211> 1356

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 48

gaagtgcagc tggtggaatc tggcggcgga ctggtcaagc ctggcggctc cctgagactg 60

tcttgcgccg cctccggctt caccttctcc acctacgcca tgtcctggggt ccgacaggcc 120

cctggaaagg gcctggagtg ggtggccacc atctccgacg ccggctccta ctcctactac 180

cccgacaacg tgaagggccg gttcaccatc tcccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgcgc cagacggggc 300

tccagatacg aagagtacta cgtgatggac tactggggcc agggcacaac cgtgaccgtg 360

tcctccgcct ccaccaaggg accctccgtg ttccctctgg ccccttccag caagtccacc 420

tctggcggca ccgccgctct gggctgcctg gtcaaggact acttcccctg ccctgtgaca 480

gtgtcctgga actccggcgc tctgacctcc ggcgtgcaca ccttccctgc cgtgctgcag 540

tcctccggcc tgtactccct gtcctccgtc gtgaccgtgc cttcctccag cctgggcacc 600

cagacctaca tctgcaacgt gaaccacaag ccctccaaca ccaaagtgga caagcgggtg 660

gaacccaagt cctgcgacaa gacccacacc tgtcctccct gccctgcccc tgagctgctg 720

ggaggccctt ccgtgttcct gttccctcca aagcccaagg acaccctgat gatctcccgg 780

acccctgaag tgacctgcgt ggtggtggcc gtgtcccacg aggatcccga agtgaagttc 840

aattggtacg tggacggcgt ggaagtgcac aatgccaaga ccaagcccag agaggaacag 900

tacaactcca cctaccgggt ggtgtccgtg ctgaccgtgc tgcaccagga ctggctgaac 960

ggcaaagagt acaagtgcaa agtgtccaac aaggccctgg ccgctcccat cgaaaagacc 1020

atctccaagg ccaagggcca gcccagagag ccccaagtgt

gaagagatga ccaagaacca agtgtccctg acctgcctcg tgaagggctt ctacccctgc 1140

gatatcgccg tggagtggga gtccaacggc cagcccgaga acaactacaa gaccacccct 1200

cccgtgctgg actccgacgg ctcattcttc ctgtactcca agctgaccgt ggacaagtcc 1260

cggtggcagc agggcaacgt gttctcctgc tccgtgatgc acgaggccct gcacaaccac 1320

tacacccaga agtccctgtc cctgagcccc ggcaag 1356

<210> 49

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 49

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 50

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 50

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 51

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 51

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 52

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 52

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 53

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 53

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 54

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 54

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 55

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 55

Ser Gln Ser Ile Ser Asn Asn

fifteen

<210> 56

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 56

Tyr Ala Ser

one

<210> 57

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 57

Ser Ser Ser Trp Leu

fifteen

<210> 58

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 58

Gln Ser Ile Ser Asn Asn

fifteen

<210> 59

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 59

Tyr Ala Ser

one

<210> 60

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 60

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 61

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 61

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser 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 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 Val Glu Pro

65 70 75 80

Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Ser Ser Trp Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 62

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 62

gagatcgtgc tgacacagtc ccctgccacc ctgtctgtgt ctcccggcga gagagtgacc 60

ctgtcctgcc gggcctccca gtccatctcc aacaacctgc actggtatca gcagaagccc 120

ggccaggccc ctcggctgct gattaagtac gcctcccaga gcatctccgg catccctgcc 180

agattctccg gctccggcag cggcaccgac ttcaccctga ccatctccag cgtggaaccc 240

gaggacttcg gcgtgtactt ctgccagcag tcctcctcct ggctgacctt cggccaggggc 300

accaagctgg aaatcaag 318

<210> 63

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 63

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser 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 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 Val Glu Pro

65 70 75 80

Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Ser Ser Trp Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 64

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 64

gagatcgtgc tgacacagtc ccctgccacc ctgtctgtgt ctcccggcga gagagtgacc 60

ctgtcctgcc gggcctccca gtccatctcc aacaacctgc actggtatca gcagaagccc 120

ggccaggccc ctcggctgct gattaagtac gcctcccaga gcatctccgg catccctgcc 180

agattctccg gctccggcag cggcaccgac ttcaccctga ccatctccag cgtggaaccc 240

gaggacttcg gcgtgtactt ctgccagcag tcctcctcct ggctgacctt cggccaggggc 300

accaagctgg aaatcaagcg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagcgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 65

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 65

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser Trp His

1.5 10

<210> 66

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 66

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 67

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 67

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 68

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 68

Ser Gly Tyr Ser Trp His

fifteen

<210> 69

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 69

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 70

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 70

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 71

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 71

Gly Tyr Ser Ile Thr Ser Gly Tyr

fifteen

<210> 72

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 72

His Ser Ser Gly Ser

fifteen

<210> 73

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 73

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 74

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 74

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser

fifteen

<210> 75

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 75

Ile His Ser Ser Gly Ser Thr

fifteen

<210> 76

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 76

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr

1.5 10

<210> 77

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 77

Asp 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 Val Ser Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 78

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 78

gacgtgcagc tgcaggaatc tggccctggc ctggtcaagc cctcccagac cctgtccctg 60

acctgcaccg tgtccggcta ctctatcacc tccggctaca gctggcactg gatccggcag 120

caccccggca agggcctgga atggatggcc cacatccact cctccggctc caccaactac 180

aaccccagcc tgaagtcccg gatcaccatc tcccgggaca cctccaagaa ccagttctcc 240

ctgaagctgt cctccgtgac cgccgctgac accgccgtgt actactgtgc cagaggcggc 300

gtgcaggcct tcgcttattg gggccaggga accctggtca ccgtgtcctc c 351

<210> 79

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 79

Asp 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 Val Ser Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 80

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 80

gacgtgcagc tgcaggaatc tggccctggc ctggtcaagc cctcccagac cctgtccctg 60

acctgcaccg tgtccggcta ctctatcacc tccggctaca gctggcactg gatccggcag 120

caccccggca agggcctgga atggatggcc cacatccact cctccggctc caccaactac 180

aaccccagcc tgaagtcccg gatcaccatc tcccgggaca cctccaagaa ccagttctcc 240

ctgaagctgt cctccgtgac cgccgctgac accgccgtgt actactgtgc cagaggcggc 300

gtgcaggcct tcgcttattg gggccaggga accctggtca ccgtgtcctc cgccagcacc 360

aagggaccct ccgtgttccc tctggcccct tccagcaagt ccacctctgg cggcaccgcc 420

gctctgggct gcctcgtgaa ggactacttc ccctgccccg tgaccgtgtc ctggaactcc 480

ggcgctctga cctccggcgt gcacaccttc cctgccgtgc tgcagtcctc cggcctgtac 540

tccctgtcca gcgtcgtgac cgtgccctcc agctctctgg gcaccagac ctacatctgc 600

aacgtgaacc acaagccctc caacaccaaa gtggacaagc gggtggaacc caagtcctgc 660

gacaagaccc acacctgtcc tccctgccct gcccctgagc tgctgggagg cccttccgtg 720

ttcctgttcc ctccaaagcc caaggacacc ctgatgatct cccggacccc tgaagtgacc 780

tgcgtggtgg tggccgtgtc ccacgaggat cccgaagtga agttcaattg gtacgtggac 840

ggcgtggaag tgcacaacgc caagaccaag cccagagagg aacagtacaa ctccacctac 900

cgggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agagtacaag 960

tgcaaagtgt ccaacaaggc cctggccgct cccatcgaaa agaccatctc caaggccaag 1020

ggccagccca gagagcccca agtgtacaca ctgcctccca gccgggaaga gatgaccaag 1080

aatcaagtgt ccctgacctg tctggtcaag ggcttctacc cctgcgatat cgccgtggag 1140

tgggagtcca acggccagcc cgagaacaac tacaagacca cccctcccgt gctggactcc 1200

gacggctcat tcttcctgta ctccaagctg accgtggaca agtcccggtg gcagcaggggc 1260

aacgtgttct cctgctccgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctgt cccctggcaa g 1341

<210> 81

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 81

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 82

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 82

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 83

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 83

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 84

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 84

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 85

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 85

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 86

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 86

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 87

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 87

Ser Ser Ser Val Ile Tyr

fifteen

<210> 88

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 88

Asp Thr Ser

one

<210> 89

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 89

Trp Ser Ser Asn Pro Leu

fifteen

<210> 90

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 90

Ser Ser Val Ile Tyr

fifteen

<210> 91

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 91

Asp Thr Ser

one

<210> 92

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 92

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 93

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 93

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Ala Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Met Ser Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Met Glu Pro Glu

65 70 75 80

Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 94

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 94

gagatcgtgc tgacacagtc ccctgccacc ctgtccgcct ctccaggcga gcgcgtgaca 60

atgtcctgct ccgcctcctc ctccgtgatc tacatgcact ggtatcagca gaagcccggc 120

caggcccctc ggcggtggat ctacgatacc tccaagctgg cctccggcgt gcccgccaga 180

ttctccggct ctggctctgg caccgactac accctgacca tctccagcat ggaacccgag 240

gacgccgccg tgtactactg cggcagtgg tcctccaacc ctctgacctt cggccaggggc 300

accaagctgg aaatcaag 318

<210> 95

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 95

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Ala Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Met Ser Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Met Glu Pro Glu

65 70 75 80

Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 96

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 96

gagatcgtgc tgacacagtc ccctgccacc ctgtccgcct ctccaggcga gcgcgtgaca 60

atgtcctgct ccgcctcctc ctccgtgatc tacatgcact ggtatcagca gaagcccggc 120

caggcccctc ggcggtggat ctacgatacc tccaagctgg cctccggcgt gcccgccaga 180

ttctccggct ctggctctgg caccgactac accctgacca tctccagcat ggaacccgag 240

gacgccgccg tgtactactg cggcagtgg tcctccaacc ctctgacctt cggccaggggc 300

accaagctgg aaatcaagcg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagcgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 97

<211> 378

<212> PROTEIN

<213> Homo sapiens

<400> 97

Met Asp Leu Gly Lys Pro Met Lys Ser Val Leu Val Val Ala Leu Leu

1.5 10 15

Val Ile Phe Gln Val Cys Leu Cys Gln Asp Glu Val Thr Asp Asp Tyr

20 25 30

Ile Gly Asp Asn Thr Val Asp Tyr Thr Leu Phe Glu Ser Leu Cys

35 40 45

Ser Lys Lys Asp Val Arg Asn Phe Lys Ala Trp Phe Leu Pro Ile Met

50 55 60

Tyr Ser Ile Ile Cys Phe Val Gly Leu Leu Gly Asn Gly Leu Val Val

65 70 75 80

Leu Thr Tyr Ile Tyr Phe Lys Arg Leu Lys Thr Met Thr Asp Thr Tyr

85 90 95

Leu Leu Asn Leu Ala Val Ala Asp Ile Leu Phe Leu Leu Thr Leu Pro

100 105 110

Phe Trp Ala Tyr Ser Ala Ala Lys Ser Trp Val Phe Gly Val His Phe

115 120 125

Cys Lys Leu Ile Phe Ala Ile Tyr Lys Met Ser Phe Phe Ser Gly Met

130 135 140

Leu Leu Leu Leu Cys Ile Ser Ile Asp Arg Tyr Val Ala Ile Val Gln

145 150 155 160

Ala Val Ser Ala His Arg His Arg Ala Arg Val Leu Leu Ile Ser Lys

165 170 175

Leu Ser Cys Val Gly Ile Trp Ile Leu Ala Thr Val Leu Ser Ile Pro

180 185 190

Glu Leu Leu Tyr Ser Asp Leu Gln Arg Ser Ser Ser Glu Gln Ala Met

195 200 205

Arg Cys Ser Leu Ile Thr Glu His Val Glu Ala Phe Ile Thr Ile Gln

210 215 220

Val Ala Gln Met Val Ile Gly Phe Leu Val Pro Leu Leu Ala Met Ser

225 230 235 240

Phe Cys Tyr Leu Val Ile Ile Arg Thr Leu Leu Gln Ala Arg Asn Phe

245 250 255

Glu Arg Asn Lys Ala Ile Lys Val Ile Ile Ala Val Val Val Val Phe

260 265 270

Ile Val Phe Gln Leu Pro Tyr Asn Gly Val Val Leu Ala Gln Thr Val

275 280 285

Ala Asn Phe Asn Ile Thr Ser Ser Thr Cys Glu Leu Ser Lys Gln Leu

290 295 300

Asn Ile Ala Tyr Asp Val Thr Tyr Ser Leu Ala Cys Val Arg Cys Cys

305 310 315 320

Val Asn Pro Phe Leu Tyr Ala Phe Ile Gly Val Lys Phe Arg Asn Asp

325 330 335

Leu Phe Lys Leu Phe Lys Asp Leu Gly Cys Leu Ser Gln Glu Gln Leu

340 345 350

Arg Gln Trp Ser Ser Cys Arg His Ile Arg Arg Ser Ser Met Ser Val

355 360 365

Glu Ala Glu Thr Thr Thr Thr Thr Phe Ser Pro

370 375

<210> 98

<211> 1134

<212> DNA

<213> Homo sapiens

<400> 98

atggacctgg ggaaaccaat gaaaagcgtg ctggtggtgg ctctccttgt cattttccag 60

gtatgcctgt gtcaagatga ggtcacggac gattacatcg gagacaacac cacagtggac 120

tacactttgt tcgagtcttt gtgctccaag aaggacgtgc ggaactttaa agcctggttc 180

ctccctatca tgtactccat catttgtttc gtgggcctac tgggcaatgg gctggtcgtg 240

ttgacctata tctatttcaa gaggctcaag accatgaccg atacctacct gctcaacctg 300

gcggtggcag acatcctctt ccctcctgacc cttcccttct gggcctacag cgcggccaag 360

tcctgggtct tcggtgtcca cttttgcaag ctcatctttg ccatctacaa gatgagcttc 420

ttcagtggca tgctcctact tctttgcatc agcattgacc gctacgtggc catcgtccag 480

gctgtctcag ctcaccgcca ccgtgcccgc gtccttctca tcagcaagct gtcctgtgtg 540

ggcatctgga tactagccac agtgctctcc atccgagc tcctgtacag tgacctccag 600

aggagcagca gtgagcaagc gatgcgatgc tctctcatca cagagcatgt ggaggccttt 660

atcaccatcc aggtggccca gatggtgatc ggctttctgg tccccctgct ggccatgagc 720

ttctgttacc ttgtcatcat ccgcaccctg ctccaggcac gcaactttga gcgcaacaag 780

gccatcaagg tgatcatcgc tgtggtcgtg gtcttcatag tcttccagct gccctacaat 840

ggggtggtcc tggccgac ggtggccaac ttcaacatca ccagtagcac ctgtgagctc 900

agtaagcaac tcaacatcgc ctacgacgtc acctacagcc tggcctgcgt ccgctgctgc 960

gtcaaccctt tcttgtacgc cttcatcggc gtcaagttcc gcaacgatct cttcaagctc 1020

ttcaaggacc tgggctgcct cagccaggag cagctccggc agtggtcttc ctgtcggcac 1080

atccggcgct cctccatgag tgtggaggcc gagaccacca ccaccttctc ccca 1134

<210> 99

<211> 378

<212> PROTEIN

<213> Macaca fascicularis

<400> 99

Met Asp Leu Gly Lys Pro Met Lys Ser Val Leu Val Val Ala Leu Leu

1.5 10 15

Val Ile Phe Gln Val Cys Leu Cys Gln Asp Glu Val Thr Asp Asp Tyr

20 25 30

Ile Gly Asp Asn Thr Val Asp Tyr Thr Leu Phe Glu Ser Leu Cys

35 40 45

Ser Lys Lys Asp Val Arg Asn Phe Lys Ala Trp Phe Leu Pro Ile Met

50 55 60

Tyr Ser Ile Ile Cys Phe Val Gly Leu Leu Gly Asn Gly Leu Val Val

65 70 75 80

Leu Thr Tyr Ile Tyr Phe Lys Arg Leu Lys Thr Met Thr Asp Thr Tyr

85 90 95

Leu Leu Asn Leu Ala Val Ala Asp Ile Leu Phe Leu Leu Thr Leu Pro

100 105 110

Phe Trp Ala Tyr Ser Ala Ala Lys Ser Trp Val Phe Gly Val His Phe

115 120 125

Cys Lys Leu Ile Phe Ala Ile Tyr Lys Met Ser Phe Phe Ser Gly Met

130 135 140

Leu Leu Leu Leu Cys Ile Ser Ile Asp Arg Tyr Val Ala Ile Val Gln

145 150 155 160

Ala Val Ser Ala His Arg His Arg Ala Arg Val Leu Leu Ile Ser Lys

165 170 175

Leu Ser Cys Val Gly Ile Trp Ile Leu Ala Thr Val Leu Ser Ile Pro

180 185 190

Glu Leu Leu Tyr Ser Gly Leu Gln Arg Ser Ser Ser Glu Gln Ala Met

195 200 205

Arg Cys Ser Leu Ile Thr Glu His Val Glu Ala Phe Ile Thr Ile Gln

210 215 220

Val Ala Gln Met Val Ile Gly Phe Leu Val Pro Leu Leu Ala Met Ser

225 230 235 240

Phe Cys Tyr Leu Val Ile Ile Arg Thr Leu Leu Gln Ala Arg Asn Phe

245 250 255

Glu Arg Asn Lys Ala Ile Lys Val Ile Ile Ala Val Val Val Val Phe

260 265 270

Ile Val Phe Gln Leu Pro Tyr Asn Gly Val Val Leu Ala Gln Thr Val

275 280 285

Ala Asn Phe Asn Ile Thr Ser Ser Thr Cys Glu Leu Ser Lys Gln Leu

290 295 300

Asn Ile Ala Tyr Asp Val Thr Tyr Ser Leu Ala Cys Val Arg Cys Cys

305 310 315 320

Val Asn Pro Phe Leu Tyr Ala Phe Ile Gly Val Lys Phe Arg Asn Asp

325 330 335

Leu Phe Lys Leu Phe Lys Asp Leu Gly Cys Leu Ser Gln Glu Gln Leu

340 345 350

Arg Gln Trp Ser Ser Cys Arg His Ile Arg Arg Ser Ser Met Ser Val

355 360 365

Glu Ala Glu Thr Thr Thr Thr Thr Phe Ser Pro

370 375

<210> 100

<211> 1134

<212> DNA

<213> Macaca fascicularis

<400> 100

atggacctgg ggaaaccaat gaaaagcgtg ctggtggtgg ctctccttgt cattttccag 60

gtatgcctgt gtcaagatga ggtcacggac gattacatcg gagacaacac cacagtggac 120

tacactttgt tcgagtcttt gtgctccaag aaggacgtgc ggaactttaa agcctggttc 180

ctccctatca tgtactccat catttgtttc gtgggcctac tgggcaatgg gctggtcgtg 240

ttgacctata tctatttcaa gaggctcaag accatgaccg atacctacct gctcaacctg 300

gcggtggcag acatcctctt ccctcctgacc cttcccttct gggcctacag cgcggccaag 360

tcctgggtct tcggtgtcca cttttgcaag ctcatctttg ccatctacaa gatgagcttc 420

ttcagtggca tgctcctact tctttgcatc agcattgacc gctacgtggc catcgtccag 480

gctgtctcag ctcaccgcca ccgtgcccgc gtccttctca tcagcaagct gtcctgtgtg 540

ggcatctgga tactagccac agtgctctcc atccgagc tcctgtacag tggcctccag 600

aggagcagca gtgagcaagc gatgcgatgc tctctcatca cagagcatgt ggaggccttt 660

atcaccatcc aggtggccca gatggtgatc ggctttctgg tccccctgct ggccatgagc 720

ttctgttacc ttgtcatcat ccgcaccctg ctccaggcac gcaactttga gcgcaacaag 780

gccatcaagg tgatcatcgc tgtggtcgtg gtcttcatag tcttccagct gccctacaat 840

ggggtggtcc tggccgac ggtggccaac ttcaacatca ccagtagcac ctgtgagctc 900

agtaagcaac tcaacatcgc ctacgacgtc acctacagcc tggcctgcgt ccgctgctgc 960

gtcaaccctt tcttgtacgc cttcatcggc gtcaagttcc gcaacgatct cttcaagctc 1020

ttcaaggacc tgggctgcct cagccaggag cagctccggc agtggtcttc ctgtcggcac 1080

atccggcgct cctccatgag tgtggaggcc gagaccacca ccaccttctc ccca 1134

<210> 101

<211> 378

<212> PROTEIN

<213> Mus sp.

<400> 101

Met Asp Pro Gly Lys Pro Arg Lys Asn Val Leu Val Val Ala Leu Leu

1.5 10 15

Val Ile Phe Gln Val Cys Phe Cys Gln Asp Glu Val Thr Asp Asp Tyr

20 25 30

Ile Gly Glu Asn Thr Val Asp Tyr Thr Leu Tyr Glu Ser Val Cys

35 40 45

Phe Lys Lys Asp Val Arg Asn Phe Lys Ala Trp Phe Leu Pro Leu Met

50 55 60

Tyr Ser Val Ile Cys Phe Val Gly Leu Leu Gly Asn Gly Leu Val Ile

65 70 75 80

Leu Thr Tyr Ile Tyr Phe Lys Arg Leu Lys Thr Met Thr Asp Thr Tyr

85 90 95

Leu Leu Asn Leu Ala Val Ala Asp Ile Leu Phe Leu Leu Ile Leu Pro

100 105 110

Phe Trp Ala Tyr Ser Glu Ala Lys Ser Trp Ile Phe Gly Val Tyr Leu

115 120 125

Cys Lys Gly Ile Phe Gly Ile Tyr Lys Leu Ser Phe Phe Ser Gly Met

130 135 140

Leu Leu Leu Leu Cys Ile Ser Ile Asp Arg Tyr Val Ala Ile Val Gln

145 150 155 160

Ala Val Ser Ala His Arg His Arg Ala Arg Val Leu Leu Ile Ser Lys

165 170 175

Leu Ser Cys Val Gly Ile Trp Met Leu Ala Leu Phe Leu Ser Ile Pro

180 185 190

Glu Leu Leu Tyr Ser Gly Leu Gln Lys Asn Ser Gly Glu Asp Thr Leu

195 200 205

Arg Cys Ser Leu Val Ser Ala Gln Val Glu Ala Leu Ile Thr Ile Gln

210 215 220

Val Ala Gln Met Val Phe Gly Phe Leu Val Pro Met Leu Ala Met Ser

225 230 235 240

Phe Cys Tyr Leu Ile Ile Ile Arg Thr Leu Leu Gln Ala Arg Asn Phe

245 250 255

Glu Arg Asn Lys Ala Ile Lys Val Ile Ile Ala Val Val Val Val Phe

260 265 270

Ile Val Phe Gln Leu Pro Tyr Asn Gly Val Val Leu Ala Gln Thr Val

275 280 285

Ala Asn Phe Asn Ile Thr Asn Ser Ser Cys Glu Thr Ser Lys Gln Leu

290 295 300

Asn Ile Ala Tyr Asp Val Thr Tyr Ser Leu Ala Ser Val Arg Cys Cys

305 310 315 320

Val Asn Pro Phe Leu Tyr Ala Phe Ile Gly Val Lys Phe Arg Ser Asp

325 330 335

Leu Phe Lys Leu Phe Lys Asp Leu Gly Cys Leu Ser Gln Glu Arg Leu

340 345 350

Arg His Trp Ser Ser Cys Arg His Val Arg Asn Ala Ser Val Ser Met

355 360 365

Glu Ala Glu Thr Thr Thr Thr Thr Phe Ser Pro

370 375

<210> 102

<211> 1134

<212> DNA

<213> Mus sp.

<400> 102

atggacccag ggaaacccag gaaaaacgtg ctggtggtgg ctctccttgt cattttccag 60

gtgtgcttct gccaagatga ggtcaccgat gactacatcg gcgagaatac cacggtggac 120

tacaccctgt acgagtcggt gtgcttcaag aaggatgtgc ggaactttaa ggcctggttc 180

ctgcctctca tgtattctgt catctgcttc gtgggcctgc tcggcaacgg gctggtgata 240

ctgacgtaca tctatttcaa gaggctcaag accatgacgg atacctacct gctcaacctg 300

gccgtggcag acatcctttt ccctcctgatt cttcccttct gggcctacag cgaagccaag 360

tcctggatct ttggcgtcta cctgtgtaag ggcatctttg gcatctataa gttaagcttc 420

ttcagcggga tgctgctgct cctatgcatc agcattgacc gctacgtagc catcgtccag 480

gccgtgtcgg ctcatcgcca ccgcgcccgc gtgcttctca tcagcaagct gtcctgtgtg 540

ggcatctgga tgctggccct cttcctctcc atcccggagc tgctctacag cggcctccag 600

aagaacagcg gcgaggacac gctgagatgc tcactggtca gtgcccaagt ggaggccttg 660

atcaccatcc aagtggccca gatggttttt gggttcctag tgcctatgct ggctatgagt 720

ttctgctacc tcattatcat ccgtaccttg ctccaggcac gcaactttga gcggaacaag 780

gccatcaagg tgatcattgc cgtggtggta gtcttcatag tcttccagct gccctacaat 840

ggggtggtcc tggctcagac ggtggccaac ttcaacatca ccaatagcag ctgcgaaacc 900

agcaagcagc tcaacattgc ctatgacgtc acctacagcc tggcctccgt ccgctgctgc 960

gtcaaccctt tcttgtatgc cttcatcggc gtcaagttcc gcagcgacct cttcaagctc 1020

ttcaaggact tgggctgtct cagccaggaa cggctccggc actggtcttc ctgccggcat 1080

gtacggaacg cgtcggtgag catggaggcg gagaccacca caaccttctc cccg 1134

<210> 103

<211> 378

<212> PROTEIN

<213> Rattus sp.

<400> 103

Met Asp Leu Gly Lys Pro Thr Lys Asn Val Leu Val Val Ala Leu Leu

1.5 10 15

Val Ile Phe Gln Val Cys Phe Cys Gln Asp Glu Val Thr Asp Asp Tyr

20 25 30

Ile Gly Glu Asn Thr Val Asp Tyr Thr Leu Tyr Glu Ser Val Cys

35 40 45

Phe Lys Lys Asp Val Arg Asn Phe Lys Ala Trp Phe Leu Pro Leu Met

50 55 60

Tyr Ser Val Ile Cys Phe Val Gly Leu Leu Gly Asn Gly Leu Val Val

65 70 75 80

Leu Thr Tyr Ile Tyr Phe Lys Arg Leu Lys Thr Met Thr Asp Thr Tyr

85 90 95

Leu Leu Asn Leu Ala Val Ala Asp Ile Leu Phe Leu Met Ile Leu Pro

100 105 110

Phe Trp Ala Tyr Ser Glu Ala Lys Ser Trp Ile Phe Gly Ala Tyr Leu

115 120 125

Cys Lys Ser Ile Phe Gly Ile Tyr Lys Leu Ser Phe Phe Ser Gly Met

130 135 140

Leu Leu Leu Leu Cys Ile Ser Ile Asp Arg Tyr Val Ala Ile Val Gln

145 150 155 160

Ala Val Ser Ala His Arg His Arg Ala Arg Val Leu Leu Ile Ser Lys

165 170 175

Leu Ser Cys Ile Gly Ile Trp Thr Leu Ala Phe Phe Leu Ser Ile Pro

180 185 190

Glu Leu Leu Tyr Ser Gly Leu Gln Lys Asn Ser Gly Glu Asp Thr Trp

195 200 205

Arg Cys Ser Leu Val Ser Ala Gln Val Glu Ala Leu Ile Ala Ile Gln

210 215 220

Val Ala Gln Met Val Val Gly Phe Val Leu Pro Met Leu Ala Met Ser

225 230 235 240

Phe Cys Tyr Leu Val Ile Ile Arg Thr Leu Leu Gln Ala Arg Asn Phe

245 250 255

Glu Arg Asn Lys Ala Ile Lys Val Ile Ile Ala Val Val Val Val Phe

260 265 270

Val Val Phe Gln Leu Pro Tyr Asn Gly Val Val Leu Ala Gln Thr Val

275 280 285

Ala Asn Phe Asn Ile Thr Asn Ser Ser Cys Glu Ala Ser Lys Gln Leu

290 295 300

Asn Ile Ala Tyr Asp Val Thr Tyr Ser Leu Ala Ser Val Arg Cys Cys

305 310 315 320

Val Asn Pro Phe Leu Tyr Ala Phe Ile Gly Val Lys Phe Arg Ser Asp

325 330 335

Leu Phe Lys Leu Phe Lys Asp Leu Gly Cys Leu Ser Gln Glu Arg Leu

340 345 350

Arg Gln Trp Ser Ser Cys Arg His Val Arg His Thr Ser Val Ser Met

355 360 365

Glu Ala Glu Thr Thr Thr Thr Thr Phe Ser Pro

370 375

<210> 104

<211> 1134

<212> DNA

<213> Rattus sp.

<400> 104

atggacctgg ggaagcccac gaaaaacgtg ctggtggtgg ctctcctggt cattttccag 60

gtgtgcttct gccaagatga ggtcacagac gactacatcg gcgagaacac caccgtggac 120

tacaccctgt atgagtcggt gtgcttcaag aaggatgtgc ggaactttaa ggcctggttc 180

ctccctctca tgtactcagt catttgcttc gtgggcctgc taggcaatgg gctggtggtg 240

ctgacataca tctatttcaa gagactgaag accatgacgg atacctacct gctcaacctg 300

gccgtggcag acatcctctt cctcatgatc cttcccttct gggcctacag cgaagccaag 360

tcctggatct ttggtgccta cctgtgtaag agcatctttg gcatctacaa gttaagcttc 420

ttcagcggga tgttgctgct cctgtgtatc agcattgacc gctatgtggc catcgtccag 480

gccgtgtcag cccaccggca ccgcgcccgc gtgcttctca tcagcaagct gtcctgtata 540

ggcatctgga cgctggcctt ttttcctttct atccctgagc tgctctacag cggcctccag 600

aagaacagcg gcgaggacac gtggagatgc tccctggtca gtgcccaagt ggaggccttg 660

atcgccatcc aagtggccca gatggttgtt gggtttgtac tgcctatgct ggctatgagt 720

ttctgctacc tggttatcat ccgcactctg ctccaggcgc gaaacttcga gcggaacaag 780

gccatcaagg tgatcatcgc tgtggtcgta gtgttcgtcg tcttccagct gccctacaat 840

ggggtggtcc tggccgac cgtggccaat ttcaacatca ccaatagcag ctgcgaagcc 900

agcaagcagc tcaacattgc ctatgacgtc acctacagcc tggcctccgt ccgctgctgt 960

gtcaaccctt tcttgtatgc cttcatcggc gtcaagttcc gcagcgacct cttcaagctc 1020

ttcaaggact tgggctgcct cagccaggaa cggctccggc agtggtcttc ctgccgccat 1080

gtacggcaca cgtccgtgag catggaggcg gagactacca ccaccttctc cccg 1134

<210> 105

<211> 378

<212> PROTEIN

<213> Homo sapiens

<400> 105

Met Asp Leu Gly Lys Pro Met Lys Ser Val Leu Val Val Ala Leu Leu

1.5 10 15

Val Ile Phe Gln Val Cys Leu Cys Gln Asp Glu Val Thr Asp Asp Tyr

20 25 30

Ile Gly Asp Asn Thr Val Asp Tyr Thr Leu Phe Glu Ser Leu Cys

35 40 45

Ser Lys Lys Asp Val Arg Asn Phe Lys Ala Trp Phe Leu Pro Ile Met

50 55 60

Tyr Ser Ile Ile Cys Phe Val Gly Leu Leu Gly Asn Gly Leu Val Val

65 70 75 80

Leu Thr Tyr Ile Tyr Phe Lys Arg Leu Lys Thr Met Thr Asp Thr Tyr

85 90 95

Leu Leu Asn Leu Ala Val Ala Asp Ile Leu Phe Leu Leu Thr Leu Pro

100 105 110

Phe Trp Ala Tyr Ser Ala Ala Lys Ser Trp Val Phe Gly Val His Phe

115 120 125

Cys Lys Leu Ile Phe Ala Ile Tyr Lys Met Ser Phe Phe Ser Gly Met

130 135 140

Leu Leu Leu Leu Cys Ile Ser Ile Asp Arg Tyr Val Ala Ile Val Gln

145 150 155 160

Ala Val Ser Ala His Arg His Arg Ala Arg Val Leu Leu Ile Ser Lys

165 170 175

Leu Ser Cys Val Gly Ile Trp Ile Leu Ala Thr Val Leu Ser Ile Pro

180 185 190

Glu Leu Leu Tyr Ser Asp Leu Gln Arg Ser Ser Ser Glu Gln Ala Met

195 200 205

Arg Cys Ser Leu Ile Thr Glu His Val Glu Ala Phe Ile Thr Ile Gln

210 215 220

Val Ala Gln Met Val Ile Gly Phe Leu Val Pro Leu Leu Ala Met Ser

225 230 235 240

Phe Cys Tyr Leu Val Ile Ile Arg Thr Leu Leu Gln Ala Arg Asn Phe

245 250 255

Glu Arg Asn Lys Ala Ile Lys Val Ile Ile Ala Val Val Val Val Phe

260 265 270

Ile Val Phe Gln Leu Pro Tyr Asn Gly Val Val Leu Ala Gln Thr Val

275 280 285

Ala Asn Phe Asn Ile Thr Ser Ser Thr Cys Glu Leu Ser Lys Gln Leu

290 295 300

Asn Ile Ala Tyr Asp Val Thr Tyr Ser Leu Ala Cys Val Arg Cys Cys

305 310 315 320

Val Asn Pro Phe Leu Tyr Ala Phe Ile Gly Val Lys Phe Arg Asn Asp

325 330 335

Leu Phe Lys Leu Phe Lys Asp Leu Gly Cys Leu Ser Gln Glu Gln Leu

340 345 350

Arg Gln Trp Ser Ser Cys Arg His Ile Arg Arg Ser Ser Met Ser Val

355 360 365

Glu Ala Glu Thr Thr Thr Thr Thr Phe Ser Pro

370 375

<210> 106

<211> 30

<212> PROTEIN

<213> Homo sapiens

<400> 106

Gln Asp Glu Val Thr Asp Asp Tyr Ile Gly Asp Asn Thr Thr Val Asp

1.5 10 15

Tyr Thr Leu Phe Glu Ser Leu Cys Ser Lys Lys Asp Val Arg

20 25 30

<210> 107

<211> 8

<212> PROTEIN

<213> Homo sapiens

<400> 107

Lys Ser Trp Val Phe Gly Val His

fifteen

<210> 108

<211> 19

<212> PROTEIN

<213> Homo sapiens

<400> 108

Tyr Ser Asp Leu Gln Arg Ser Ser Ser Glu Gln Ala Met Arg Cys Ser

1.5 10 15

Leu Ile Thr

<210> 109

<211> 7

<212> PROTEIN

<213> Homo sapiens

<400> 109

Phe Asn Ile Thr Ser Ser Thr

fifteen

<210> 110

<211> 19

<212> PROTEIN

<213> Homo sapiens

<400> 110

Tyr Ser Asp Leu Gln Arg Ser Ser Ser Glu Gln Ala Met Arg Ser Ser

1.5 10 15

Leu Ile Thr

<210> 111

<211> 216

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 111

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 Leu Trp Val Arg Gln Ala Pro Glu Lys Gly Leu Glu Trp Ile

35 40 45

Ala Tyr Ile Ser Ser Gly Ser Ser Thr Ile Tyr Tyr Ala Asp Arg Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Phe

65 70 75 80

Leu Gln Met Thr Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ser Thr Gly Thr Phe Ala Tyr Trp Gly Gln Gly Thr Pro Val Thr Val

100 105 110

Ser Ser Ala Lys Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly

115 120 125

Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly Cys Leu Val Lys

130 135 140

Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Ser Leu

145 150 155 160

Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr

165 170 175

Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser Glu

180 185 190

Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val Asp

195 200 205

Lys Lys Ile Val Pro Arg Asp Cys

210 215

<210> 112

<211> 219

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 112

Asp Val Val Met Thr Gln Asn Pro Leu Ser Leu Pro Val Ser Leu Gly

1.5 10 15

Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Ile Tyr Asn

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp Tyr Arg Gln Lys Pro Gly Gln Ser

35 40 45

Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser

85 90 95

Thr His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu

115 120 125

Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe

130 135 140

Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg

145 150 155 160

Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu

180 185 190

Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser

195 200 205

Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys

210 215

<210> 113

<211> 265

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 113

Gln Asp Glu Val Thr Asp Asp Tyr Ile Gly Asp Asn Thr Thr Val Asp

1.5 10 15

Tyr Thr Leu Phe Glu Ser Leu Cys Ser Lys Lys Asp Val Arg Glu Val

20 25 30

Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly Ser Leu

35 40 45

Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr Gly Met

50 55 60

Leu Trp Val Arg Gln Ala Pro Glu Lys Gly Leu Glu Trp Ile Ala Tyr

65 70 75 80

Ile Ser Ser Gly Ser Ser Thr Ile Tyr Tyr Ala Asp Arg Val Lys Gly

85 90 95

Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Phe Leu Gln

100 105 110

Met Thr Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys Ser Thr

115 120 125

Gly Thr Tyr Ser Asp Leu Gln Arg Ser Ser Ser Glu Gln Ala Met Arg

130 135 140

Ser Ser Leu Ile Thr Phe Ala Tyr Trp Gly Gln Gly Thr Pro Val Thr

145 150 155 160

Val Ser Ser Ala Lys Thr Pro Pro Ser Val Tyr Pro Leu Ala Pro

165 170 175

Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly Cys Leu Val

180 185 190

Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Ser

195 200 205

Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp Leu

210 215 220

Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro Ser

225 230 235 240

Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys Val

245 250 255

Asp Lys Lys Ile Val Pro Arg Asp Cys

260 265

<210> 114

<211> 795

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 114

caagatgagg tcacggacga ttacatcggga gacaacacca cagtggacta cactttgttc 60

gagtctttgt gctccaagaa ggacgtgcgg gaggtgcagc tggtggagtc tggtggtggt 120

ctggtcaagc ctggaggttc ctgaaactg agttgtgccg catctgggtt tacattctct 180

gactacggaa tgctgtgggt gaggcaggca ccagagaagg gcctggaatg gatcgcttat 240

atttccagcg gatctagtac tatctactat gcagacaggg tcaagggccg gttcaccatt 300

agcagagata acgccaaaaa taccctgttt ctgcagatga catcactgag gtccgaggat 360

accgctatgt attattgctc cacagggact tacagtgacc tccagaggag cagcagtgag 420

caagcgatgc gatcctctct catcacattt gcttactggg gacaggggac acccgtgacc 480

gtcagctcag ccaagaccac cccccccagc gtgtaccctc tggcccctgg ctctgccgcc 540

cagaccaaca gcatggtgac cctgggctgc ctggtgaagg gctacttccc cgagcccgtg 600

accgtgacct ggaacagcgg cagcctgagc agcggcgtgc acaccttccc cgccgtgctg 660

cagagcgacc tgtacaccct gagcagctct gtgaccgtgc ccagcagcac ctggcccagc 720

gagaccgtga catgcaacgt ggcccacccc gccagctcca ccaaggtgga caagaaaatc 780

gtgccccggg actgc 795

<210> 115

<211> 656

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 115

atgtcgtgat gactcagaat ccactgtccc tgcctgtgtc cctgggcgat caggcttcca 60

ttagctgtcg ttcctctcag tccctgatct acaacaatgg taacacctac ctgcactggt 120

atagacagaa gcccggccag tcccctaagc tgctgatcta caaagtgagt aataggttct 180

caggagtccc agaccggttt tccggcagcg gatctgggac cgatttcaca ctgaaaatct 240

ctagggtgga ggccgaagac ctgggcgtct acttttgtag tcagagcact cacgtcccct 300

tcaccttcgg cagcggaaca aaactggaaa tcaagcgcgc tgatgccgcc cctaccgtga 360

gcatcttccc ccccagcagc gagcagctga ccagcggcgg agccagcgtg gtgtgcttcc 420

tgaacaactt ctaccccaag gacatcaacg tgaagtggaa gatcgacggc agcgagcggc 480

agaacggcgt gctgaacagc tggaccgacc aggacagcaa ggactccacc tacagcatga 540

gcagcaccct gaccctgacc aaggacgagt acgagcggca caacagctac acctgcgagg 600

ccacccacaa gaccagcacc agccccatcg tgaagagctt caaccggaac gagtgc 656

<210> 116

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 116

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser Trp His

1.5 10

<210> 117

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 117

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 118

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 118

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 119

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 119

Ser Gly Tyr Ser Trp His

fifteen

<210> 120

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 120

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 121

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 121

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 122

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 122

Gly Tyr Ser Ile Thr Ser Gly Tyr

fifteen

<210> 123

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 123

His Ser Ser Gly Ser

fifteen

<210> 124

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 124

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 125

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 125

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser

fifteen

<210> 126

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 126

Ile His Ser Ser Gly Ser Thr

fifteen

<210> 127

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 127

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr

1.5 10

<210> 128

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 128

Asp Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln

1.5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Ser Ile Ile Arg Asp Thr Ser Lys Asn Leu Phe Phe

65 70 75 80

Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala

115

<210> 129

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 129

gatgtgcagc ttcaggagtc aggacctgac ctggtgaaac cttctcagtc actttcactc 60

acctgcactg tcactggcta ctccatcacc agtggttata gctggcactg gatccggcag 120

tttcggaa acaaactgga gtggatggcc cacatccact ccagtggtag cactaactac 180

aacccatctc tcaaaagtcg catctctatc attcgagaca catccaagaa cctgttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagagggggg 300

gtacaggcct ttgcttactg gggccaaggg actctggtca ctgtctctgc a 351

<210> 130

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 130

Asp Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln

1.5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Ser Ile Ile Arg Asp Thr Ser Lys Asn Leu Phe Phe

65 70 75 80

Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 131

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 131

gatgtgcagc ttcaggagtc aggacctgac ctggtgaaac cttctcagtc actttcactc 60

acctgcactg tcactggcta ctccatcacc agtggttata gctggcactg gatccggcag 120

tttcggaa acaaactgga gtggatggcc cacatccact ccagtggtag cactaactac 180

aacccatctc tcaaaagtcg catctctatc attcgagaca catccaagaa cctgttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagagggggg 300

gtacaggcct ttgcttactg gggccaaggg actctggtca ctgtctctgc agctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc cccgagcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggacgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctgccagcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 132

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 132

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 133

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 133

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 134

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 134

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 135

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 135

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 136

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 136

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 137

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 137

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 138

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 138

Ser Ser Ser Val Ile Tyr

fifteen

<210> 139

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 139

Asp Thr Ser

one

<210> 140

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 140

Trp Ser Ser Asn Pro Leu

fifteen

<210> 141

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 141

Ser Ser Val Ile Tyr

fifteen

<210> 142

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 142

Asp Thr Ser

one

<210> 143

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 143

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 144

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 144

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly

1.5 10 15

Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Ala Gly Thr Thr Leu Glu Leu Lys

100 105

<210> 145

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 145

caaattgtcc tcacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc 60

atgacctgca gtgccagttc aagtgtaatt tacatgcact ggtaccagca gaagtcaggc 120

acctccccca aaagatggat ttatgacaca tccaaactgg cttctggagt ccctgctcgc 180

ttcagtggta gtgggtctgg gacctcttac tctctcacaa tcagcagcat ggaggctgaa 240

gatgctgcca cttattactg ccagcagtgg agtagtaacc cgctcacgtt cggtgctggg 300

accacgttgg agctgaaa 318

<210> 146

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 146

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly

1.5 10 15

Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Ala Gly Thr Thr Leu Glu Leu Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 147

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 147

caaattgtcc tcacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc 60

atgacctgca gtgccagttc aagtgtaatt tacatgcact ggtaccagca gaagtcaggc 120

acctccccca aaagatggat ttatgacaca tccaaactgg cttctggagt ccctgctcgc 180

ttcagtggta gtgggtctgg gacctcttac tctctcacaa tcagcagcat ggaggctgaa 240

gatgctgcca cttattactg ccagcagtgg agtagtaacc cgctcacgtt cggtgctggg 300

accacgttgg agctgaaacg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 148

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 148

Gly Phe Thr Phe Ser Thr Tyr Ala Met Ser

1.5 10

<210> 149

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 149

Thr Ile Ser Asp Gly Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 150

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 150

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 151

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 151

Thr Tyr Ala Met Ser

fifteen

<210> 152

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 152

Thr Ile Ser Asp Gly Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 153

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 153

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 154

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 154

Gly Phe Thr Phe Ser Thr Tyr

fifteen

<210> 155

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 155

Ser Asp Gly Gly Ser Tyr

fifteen

<210> 156

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 156

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 157

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 157

Gly Phe Thr Phe Ser Thr Tyr Ala

fifteen

<210> 158

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 158

Ile Ser Asp Gly Gly Ser Tyr Ser

fifteen

<210> 159

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 159

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10 15

<210> 160

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 160

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 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 Asp Gly Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr

65 70 75 80

Leu Gln Met Ser His Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 161

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 161

gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt acctatgcca tgtcttgggt tcgccagact 120

ccggaaaaga ggctggagtg ggtcgcaacc attagtgatg gtggtagtta ttcgtactat 180

ccagacaatg taaagggccg attcaccatc tccagagaca atgccaagaa caacctatac 240

ctgcaaatga gccatctgaa gtctgaggac acagccatgt attactgtgc aagacgaggt 300

agtaggtacg aagagtacta tgttatggac tactggggtc aaggaacctc agtcaccgtc 360

tcctca 366

<210> 162

<211> 452

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 162

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 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 Asp Gly Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr

65 70 75 80

Leu Gln Met Ser His Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 163

<211> 1356

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 163

gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt acctatgcca tgtcttgggt tcgccagact 120

ccggaaaaga ggctggagtg ggtcgcaacc attagtgatg gtggtagtta ttcgtactat 180

ccagacaatg taaagggccg attcaccatc tccagagaca atgccaagaa caacctatac 240

ctgcaaatga gccatctgaa gtctgaggac acagccatgt attactgtgc aagacgaggt 300

agtaggtacg aagagtacta tgttatggac tactggggtc aaggaacctc agtcaccgtc 360

tcctcagcta gcaccaaggg cccaagtgtg tttcccctgg cccccagcag caagtctact 420

tccggcggaa ctgctgccct gggttgcctg gtgaaggact acttccccga gcccgtgaca 480

gtgtcctgga actctggggc tctgacttcc ggcgtgcaca ccttccccgc cgtgctgcag 540

agcagcggcc tgtacagcct gagcagcgtg gtgacagtgc cctccagctc tctgggaacc 600

cagacctata tctgcaacgt gaaccacaag cccagcaaca ccaaggtgga caagagagtg 660

gagcccaaga gctgcgacaa gacccacacc tgccccccct gcccagctcc agaactgctg 720

ggagggcctt ccgtgttcct gttccccccc aagcccaagg acaccctgat gatcagcagg 780

acccccgagg tgacctgcgt ggtggtggac gtgtcccacg aggacccaga ggtgaagttc 840

aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcccag agaggagcag 900

tacaacagca cctacagggt ggtgtccgtg ctgaccgtgc tgcaccagga ctggctgaac 960

ggcaaagaat acaagtgcaa agtctccaac aaggccctgc cagccccaat cgaaaagaca 1020

atcagcaagg ccaagggcca gccacgggag ccccaggtgt acaccctgcc ccccagccgg 1080

gaggagatga ccaagaacca ggtgtccctg acctgtctgg tgaagggctt ctaccccagc 1140

gatatcgccg tggagtggga gagcaacggc cagcccgaga acaactacaa gaccaccccc 1200

ccagtgctgg acagcgacgg cagcttcttc ctgtacagca agctgaccgt ggacaagtcc 1260

aggtggcagc agggcaacgt gttcagctgc agcgtgatgc acgaggccct gcacaaccac 1320

tacacccaga agtccctgag cctgagcccc ggcaag 1356

<210> 164

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 164

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 165

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 165

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 166

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 166

Gln Gln Ser Asn Ser Trp Leu Thr

fifteen

<210> 167

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 167

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 168

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 168

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 169

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 169

Gln Gln Ser Asn Ser Trp Leu Thr

fifteen

<210> 170

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 170

Ser Gln Ser Ile Ser Asn Asn

fifteen

<210> 171

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 171

Tyr Ala Ser

one

<210> 172

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 172

Ser Asn Ser Trp Leu

fifteen

<210> 173

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 173

Gln Ser Ile Ser Asn Asn

fifteen

<210> 174

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 174

Tyr Ala Ser

one

<210> 175

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 175

Gln Gln Ser Asn Ser Trp Leu Thr

fifteen

<210> 176

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 176

Asp Ile 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 Ser Ile Ser Asn Asn

20 25 30

Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Lys 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 Ser Val Glu Thr

65 70 75 80

Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ser Asn Ser Trp Leu Thr

85 90 95

Phe Gly Ala Gly Thr Lys Leu Gly Leu Lys

100 105

<210> 177

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 177

gatattgtgc taactcagtc tccagccacc ctgtctgtga ctccaggaga tagcgtcagt 60

ctttcctgca gggccagcca aagtattagc aacaacctac actggtatca acagaaatca 120

catgagtctc caaaacttct catcaagtat gcttcccagt ccatctctgg gatcccctcc 180

aggttcagtg gcagtggatc agggacagat ttcactctca gtatcaacag tgtggagact 240

gaagattttg gaatgtattt ctgtcaacag agtaacagct ggctcacgtt cggtgctggg 300

accaagctggggctgaaa 318

<210> 178

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 178

Asp Ile 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 Ser Ile Ser Asn Asn

20 25 30

Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Lys 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 Ser Val Glu Thr

65 70 75 80

Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ser Asn Ser Trp Leu Thr

85 90 95

Phe Gly Ala Gly Thr Lys Leu Gly Leu Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 179

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 179

gatattgtgc taactcagtc tccagccacc ctgtctgtga ctccaggaga tagcgtcagt 60

ctttcctgca gggccagcca aagtattagc aacaacctac actggtatca acagaaatca 120

catgagtctc caaaacttct catcaagtat gcttcccagt ccatctctgg gatcccctcc 180

aggttcagtg gcagtggatc agggacagat ttcactctca gtatcaacag tgtggagact 240

gaagattttg gaatgtattt ctgtcaacag agtaacagct ggctcacgtt cggtgctggg 300

accaagctgg ggctgaaacg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 180

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 180

Gly Phe Thr Phe Ser Tyr Ala Met Ser

1.5 10

<210> 181

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 181

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 182

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 182

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 183

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 183

Ser Tyr Ala Met Ser

fifteen

<210> 184

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 184

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 185

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 185

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 186

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 186

Gly Phe Thr Phe Ser Ser Tyr

fifteen

<210> 187

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 187

Ser Ser Gly Gly Ser Phe

fifteen

<210> 188

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 188

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 189

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 189

Gly Phe Thr Phe Ser Ser Tyr Ala

fifteen

<210> 190

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 190

Ile Ser Ser Gly Gly Ser Phe Thr

fifteen

<210> 191

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 191

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 192

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 192

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 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Ala Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 193

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 193

gaagtgatgc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt agctatgcca tgtcttggat tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc atcagtagtg gtggtagttt cacctactat 180

ccagacagtg tgaaggggcg attcaccatt tctagagaca atgtcaagaa caccctgtac 240

ctgcaaatga gcagtctgag gtctgaagac acggccatgt attactgtgc aagacgggct 300

tctacggtag taggtacgga cttcgatgtc tggggcgcag ggaccacggt caccgtctcc 360

tca 363

<210> 194

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 194

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 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Ala Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 195

<211> 1353

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 195

gaagtgatgc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt agctatgcca tgtcttggat tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc atcagtagtg gtggtagttt cacctactat 180

ccagacagtg tgaaggggcg attcaccatt tctagagaca atgtcaagaa caccctgtac 240

ctgcaaatga gcagtctgag gtctgaagac acggccatgt attactgtgc aagacgggct 300

tctacggtag taggtacgga cttcgatgtc tggggcgcag ggaccacggt caccgtctcc 360

tcagctagca ccaagggccc aagtgtgttt cccctggccc ccagcagcaa gtctacttcc 420

ggcggaactg ctgccctgggg ttgcctggtg aaggactact tccccgagcc cgtgacagtg 480

tcctggaact ctggggctct gacttccggc gtgcacacct tccccgccgt gctgcagagc 540

agcggcctgt acagcctgag cagcgtggtg acagtgccct ccagctctct gggaacccag 600

acctatatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gagagtggag 660

cccaagagct gcgacaagac ccacacctgc cccccctgcc cagctccaga actgctggga 720

gggccttccg tgttcctgtt cccccccaag cccaaggaca ccctgatgat cagcaggacc 780

cccgaggtga cctgcgtggt ggtggacgtg tcccacgagg acccagaggt gaagttcaac 840

tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcccagaga ggagcagtac 900

aacagcacct acagggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 960

aaagaataca agtgcaaagt ctccaacaag gccctgccag ccccaatcga aaagacaatc 1020

1080

gagatgacca agaaccaggt gtccctgacc tgtctggtga agggcttcta ccccagcgat 1140

atcgccgtgg agtgggagag caacggccag cccgagaaca actacaagac caccccccca 1200

1260

tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1320

acccagaagt ccctgagcct gagccccggc aag 1353

<210> 196

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 196

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 197

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 197

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 198

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 198

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 199

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 199

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 200

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 200

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 201

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 201

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 202

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 202

Ser Gln Asp Ile Gly Ser Ser

fifteen

<210> 203

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 203

Ala Thr Ser

one

<210> 204

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 204

Tyr Ala Ser Ser Pro

fifteen

<210> 205

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 205

Gln Asp Ile Gly Ser Ser

fifteen

<210> 206

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 206

Ala Thr Ser

one

<210> 207

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 207

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 208

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 208

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly

1.5 10 15

Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 209

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 209

gacatccaga tgacccagtc tccatcctcc ttatctgcct ctctgggaga aagagtcagt 60

ctcacttgtc gggcaagtca ggacattggt agtagcttaa actggcttca gcaggaacca 120

gatggaacta ttaaacgcct gatctatgcc acatccagtt tagattctgg tgtccccaaa 180

aggttcagtg gcagtaggtc tgggtcagat tattctctca ccatcagcag ccttgagtct 240

gaagattttg tagtctatta ctgtctacaa tatgctagtt cgcctccgac gttcggtgga 300

ggcaccaagc tggaaatcaa a 321

<210> 210

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 210

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly

1.5 10 15

Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 211

<211> 642

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 211

gacatccaga tgacccagtc tccatcctcc ttatctgcct ctctgggaga aagagtcagt 60

ctcacttgtc gggcaagtca ggacattggt agtagcttaa actggcttca gcaggaacca 120

gatggaacta ttaaacgcct gatctatgcc acatccagtt tagattctgg tgtccccaaa 180

aggttcagtg gcagtaggtc tgggtcagat tattctctca ccatcagcag ccttgagtct 240

gaagattttg tagtctatta ctgtctacaa tatgctagtt cgcctccgac gttcggtgga 300

ggcaccaagc tggaaatcaa acgtacggtg gccgctccca gcgtgttcat cttccccccc 360

agcgacgagc agctgaagag tggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420

ccccgggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480

gagagcgtca ccgagcagga cagcaaggac tccacctaca gcctgagcag caccctgacc 540

ctgagcaagg ccgactacga gaagcataag gtgtacgcct gcgaggtgac ccaccagggc 600

ctgtccagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642

<210> 212

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 212

Gly Phe Thr Phe Ser Asn Phe Ala Met Ser

1.5 10

<210> 213

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 213

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 214

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 214

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 215

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 215

Ser Asn Phe Ala Met Ser

fifteen

<210> 216

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 216

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 217

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 217

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 218

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 218

Gly Phe Thr Phe Ser Asn Phe

fifteen

<210> 219

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 219

Ser Thr Gly Gly Thr Tyr

fifteen

<210> 220

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 220

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 221

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 221

Gly Phe Thr Phe Ser Asn Phe Ala

fifteen

<210> 222

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 222

Ile Ser Thr Gly Gly Thr Tyr Thr

fifteen

<210> 223

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 223

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys

1.5 10

<210> 224

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 224

Glu Val His 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 Asn Phe

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 Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser

115

<210> 225

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 225

gaagtgcatc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt aactttgcca tgtcttgggt tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc attagtactg gtggtactta cacctactat 180

ccagacagtg tgaagggtcg attcaccatc tccagagaca atgccaagaa aaccctgtac 240

ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtac aagacggggg 300

tacgacggcg tggacaaatg gggccaaggc accactctca cagtctcctc a 351

<210> 226

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 226

Glu Val His 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 Asn Phe

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 Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 227

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 227

gaagtgcatc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt aactttgcca tgtcttgggt tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc attagtactg gtggtactta cacctactat 180

ccagacagtg tgaagggtcg attcaccatc tccagagaca atgccaagaa aaccctgtac 240

ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtac aagacggggg 300

tacgacggcg tggacaaatg gggccaaggc accactctca cagtctcctc agctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc cccgagcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggacgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctgccagcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 228

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 228

Lys Ser Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 229

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 229

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 230

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 230

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 231

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 231

Lys Ser Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 232

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 232

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 233

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 233

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 234

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 234

Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr

1.5 10

<210> 235

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 235

Leu Val Ser

one

<210> 236

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 236

Gly Thr His Phe Pro Gln

fifteen

<210> 237

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 237

Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr

1.5 10

<210> 238

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 238

Leu Val Ser

one

<210> 239

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 239

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 240

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 240

Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 241

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 241

gatgttgtga tgacccagac tccactcact ttgtcggtta ccattggaca accagcctcc 60

atctcttgca agtcaggtca gagcctctta gatagtgatg gaaagacata tttgaattgg 120

tttttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttcct 300

cagacgttcg gtggaggcac caagctggaa atcaaa 336

<210> 242

<211> 219

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 242

Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 243

<211> 657

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 243

gatgttgtga tgacccagac tccactcact ttgtcggtta ccattggaca accagcctcc 60

atctcttgca agtcaggtca gagcctctta gatagtgatg gaaagacata tttgaattgg 120

tttttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttcct 300

cagacgttcg gtggaggcac caagctggaa atcaaacgta cggtggccgc tcccagcgtg 360

ttcatcttcc cccccagcga cgagcagctg aagagtggca ccgccagcgt ggtgtgcctg 420

ctgaacaact tctacccccg ggaggccaag gtgcagtgga aggtggacaa cgccctgcag 480

agcggcaaca gccaggagag cgtcaccgag caggacagca aggactccac ctacagcctg 540

agcagcaccc tgaccctgag caaggccgac tacgagaagc ataaggtgta cgcctgcgag 600

gtgacccacc agggcctgtc cagccccgtg accaagagct tcaacagggg cgagtgc 657

<210> 244

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 244

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser Trp His

1.5 10

<210> 245

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 245

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 246

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 246

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 247

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 247

Ser Gly Tyr Ser Trp His

fifteen

<210> 248

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 248

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 249

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 249

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 250

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 250

Gly Tyr Ser Ile Thr Ser Gly Tyr

fifteen

<210> 251

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 251

His Ser Ser Gly Ser

fifteen

<210> 252

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 252

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 253

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 253

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser

fifteen

<210> 254

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 254

Ile His Ser Ser Gly Ser Thr

fifteen

<210> 255

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 255

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr

1.5 10

<210> 256

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 256

Asp Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln

1.5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Ser Ile Ile Arg Asp Thr Ser Lys Asn Leu Phe Phe

65 70 75 80

Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala

115

<210> 257

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 257

gatgtgcagc ttcaggagtc aggacctgac ctggtgaaac cttctcagtc actttcactc 60

acctgcactg tcactggcta ctccatcacc agtggttata gctggcactg gatccggcag 120

tttcggaa acaaactgga gtggatggcc cacatccact ccagtggtag cactaactac 180

aacccatctc tcaaaagtcg catctctatc attcgagaca catccaagaa cctgttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagagggggg 300

gtacaggcct ttgcttactg gggccaaggg actctggtca ctgtctctgc a 351

<210> 258

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 258

Asp Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln

1.5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Ser Ile Ile Arg Asp Thr Ser Lys Asn Leu Phe Phe

65 70 75 80

Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 259

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 259

gatgtgcagc ttcaggagtc aggacctgac ctggtgaaac cttctcagtc actttcactc 60

acctgcactg tcactggcta ctccatcacc agtggttata gctggcactg gatccggcag 120

tttcggaa acaaactgga gtggatggcc cacatccact ccagtggtag cactaactac 180

aacccatctc tcaaaagtcg catctctatc attcgagaca catccaagaa cctgttcttc 240

ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagagggggg 300

gtacaggcct ttgcttactg gggccaaggg actctggtca ctgtctctgc agctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc ccctgtcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggacgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctgccagcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc cctgtgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 260

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 260

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 261

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 261

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 262

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 262

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 263

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 263

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 264

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 264

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 265

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 265

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 266

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 266

Ser Ser Ser Val Ile Tyr

fifteen

<210> 267

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 267

Asp Thr Ser

one

<210> 268

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 268

Trp Ser Ser Asn Pro Leu

fifteen

<210> 269

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 269

Ser Ser Val Ile Tyr

fifteen

<210> 270

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 270

Asp Thr Ser

one

<210> 271

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 271

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 272

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 272

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly

1.5 10 15

Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Ala Gly Thr Thr Leu Glu Leu Lys

100 105

<210> 273

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 273

caaattgtcc tcacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc 60

atgacctgca gtgccagttc aagtgtaatt tacatgcact ggtaccagca gaagtcaggc 120

acctccccca aaagatggat ttatgacaca tccaaactgg cttctggagt ccctgctcgc 180

ttcagtggta gtgggtctgg gacctcttac tctctcacaa tcagcagcat ggaggctgaa 240

gatgctgcca cttattactg ccagcagtgg agtagtaacc cgctcacgtt cggtgctggg 300

accacgttgg agctgaaa 318

<210> 274

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 274

Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Pro Gly

1.5 10 15

Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Ala Gly Thr Thr Leu Glu Leu Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 275

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 275

caaattgtcc tcacccagtc tccagcaatc atgtctgcat ctccagggga gaaggtcacc 60

atgacctgca gtgccagttc aagtgtaatt tacatgcact ggtaccagca gaagtcaggc 120

acctccccca aaagatggat ttatgacaca tccaaactgg cttctggagt ccctgctcgc 180

ttcagtggta gtgggtctgg gacctcttac tctctcacaa tcagcagcat ggaggctgaa 240

gatgctgcca cttattactg ccagcagtgg agtagtaacc cgctcacgtt cggtgctggg 300

accacgttgg agctgaaacg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 276

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 276

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser Trp His

1.5 10

<210> 277

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 277

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 278

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 278

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 279

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 279

Ser Gly Tyr Ser Trp His

fifteen

<210> 280

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 280

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 281

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 281

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 282

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 282

Gly Tyr Ser Ile Thr Ser Gly Tyr

fifteen

<210> 283

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 283

His Ser Ser Gly Ser

fifteen

<210> 284

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 284

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 285

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 285

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser

fifteen

<210> 286

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 286

Ile His Ser Ser Gly Ser Thr

fifteen

<210> 287

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 287

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr

1.5 10

<210> 288

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 288

Asp 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 Val Ser Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 289

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 289

gacgtgcagc tgcaggaatc tggccctggc ctggtgaaac cctcccagac cctgtccctg 60

acctgcaccg tgtccggcta ctccatcacc tccggctaca gctggcactg gatccggcag 120

caccccggca agggcctgga atggatggcc cacatccact cctccggctc caccaactac 180

aaccccagcc tgaagtccag aatcaccatc agccgggaca cctccaagaa ccagttctcc 240

ctgaagctgt cctccgtgac cgccgctgac accgccgtgt actactgtgc cagaggcggc 300

gtgcaggcct tcgcttattg gggccagggc accctggtga cagtgtcctc c 351

<210> 290

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 290

Asp 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 Val Ser Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 291

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 291

60

acctgcaccg tgtccggcta ctccatcacc tccggctaca gctggcactg gatccggcag 120

caccccggca agggcctgga atggatggcc cacatccact cctccggctc caccaactac 180

aaccccagcc tgaagtccag aatcaccatc agccgggaca cctccaagaa ccagttctcc 240

ctgaagctgt cctccgtgac cgccgctgac accgccgtgt actactgtgc cagaggcggc 300

gtgcaggcct tcgcttattg gggccagggc accctggtga cagtgtcctc cgctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc ccctgtcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc cctgtgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 292

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 292

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 293

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 293

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 294

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 294

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 295

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 295

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 296

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 296

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 297

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 297

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 298

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 298

Ser Ser Ser Val Ile Tyr

fifteen

<210> 299

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 299

Asp Thr Ser

one

<210> 300

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 300

Trp Ser Ser Asn Pro Leu

fifteen

<210> 301

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 301

Ser Ser Val Ile Tyr

fifteen

<210> 302

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 302

Asp Thr Ser

one

<210> 303

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 303

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 304

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 304

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Ala Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Met Ser Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Met Glu Pro Glu

65 70 75 80

Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 305

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 305

gagatcgtgc tgacccagtc ccctgccacc ctgtctgcta gccctggcga gcgcgtgaca 60

atgtcctgct ccgcctcctc ctccgtgatc tacatgcact ggtatcagca gaagcccggc 120

caggcccctc ggcggtggat ctacgatacc tccaagctgg cctccggcgt gcccgccaga 180

ttctccggct ctggctctgg caccgactac accctgacca tctccagcat ggaacccgag 240

gacgccgccg tgtactactg cggcagtgg tcctccaacc ccctgacctt cggccaggggc 300

accaagctgg aaatcaag 318

<210> 306

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 306

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Ala Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Met Ser Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Met Glu Pro Glu

65 70 75 80

Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 307

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 307

gagatcgtgc tgacccagtc ccctgccacc ctgtctgcta gccctggcga gcgcgtgaca 60

atgtcctgct ccgcctcctc ctccgtgatc tacatgcact ggtatcagca gaagcccggc 120

caggcccctc ggcggtggat ctacgatacc tccaagctgg cctccggcgt gcccgccaga 180

ttctccggct ctggctctgg caccgactac accctgacca tctccagcat ggaacccgag 240

gacgccgccg tgtactactg cggcagtgg tcctccaacc ccctgacctt cggccaggggc 300

accaagctgg aaatcaagcg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 308

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 308

Gly Phe Thr Phe Ser Thr Tyr Ala Met Ser

1.5 10

<210> 309

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 309

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 310

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 310

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 311

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 311

Thr Tyr Ala Met Ser

fifteen

<210> 312

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 312

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 313

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 313

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 314

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 314

Gly Phe Thr Phe Ser Thr Tyr

fifteen

<210> 315

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 315

Ser Asp Ala Gly Ser Tyr

fifteen

<210> 316

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 316

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 317

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 317

Gly Phe Thr Phe Ser Thr Tyr Ala

fifteen

<210> 318

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 318

Ile Ser Asp Ala Gly Ser Tyr Ser

fifteen

<210> 319

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 319

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10 15

<210> 320

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 320

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 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 Asp Gly Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr

65 70 75 80

Leu Gln Met Ser His Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Glyn Gly Thr Ser Val Thr Val Ser Ser

115 120

<210> 321

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 321

gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt acctatgcca tgtcttgggt tcgccagact 120

ccggaaaaga ggctggagtg ggtcgcaacc attagtgatg gtggtagtta ttcgtactat 180

ccagacaatg taaagggccg attcaccatc tccagagaca atgccaagaa caacctatac 240

ctgcaaatga gccatctgaa gtctgaggac acagccatgt attactgtgc aagacgaggt 300

agtaggtacg aagagtacta tgttatggac tactggggtc aaggaacctc agtcaccgtc 360

tcctca 366

<210> 322

<211> 452

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 322

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 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 Asp Gly Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr

65 70 75 80

Leu Gln Met Ser His Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 323

<211> 1356

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 323

gaagtgcagc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt acctatgcca tgtcttgggt tcgccagact 120

ccggaaaaga ggctggagtg ggtcgcaacc attagtgatg gtggtagtta ttcgtactat 180

ccagacaatg taaagggccg attcaccatc tccagagaca atgccaagaa caacctatac 240

ctgcaaatga gccatctgaa gtctgaggac acagccatgt attactgtgc aagacgaggt 300

agtaggtacg aagagtacta tgttatggac tactggggtc aaggaacctc agtcaccgtc 360

tcctcagcta gcaccaaggg cccaagtgtg tttcccctgg cccccagcag caagtctact 420

tccggcggaa ctgctgccct gggttgcctg gtgaaggact acttcccctg tcccgtgaca 480

gtgtcctgga actctggggc tctgacttcc ggcgtgcaca ccttccccgc cgtgctgcag 540

agcagcggcc tgtacagcct gagcagcgtg gtgacagtgc cctccagctc tctgggaacc 600

cagacctata tctgcaacgt gaaccacaag cccagcaaca ccaaggtgga caagagagtg 660

gagcccaaga gctgcgacaa gacccacacc tgccccccct gcccagctcc agaactgctg 720

ggagggcctt ccgtgttcct gttccccccc aagcccaagg acaccctgat gatcagcagg 780

acccccgagg tgacctgcgt ggtggtggac gtgtcccacg aggacccaga ggtgaagttc 840

aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcccag agaggagcag 900

tacaacagca cctacagggt ggtgtccgtg ctgaccgtgc tgcaccagga ctggctgaac 960

ggcaaagaat acaagtgcaa agtctccaac aaggccctgc cagccccaat cgaaaagaca 1020

atcagcaagg ccaagggcca gccacgggag ccccaggtgt acaccctgcc ccccagccgg 1080

gaggagatga ccaagaacca ggtgtccctg acctgtctgg tgaagggctt ctacccctgt 1140

gatatcgccg tggagtggga gagcaacggc cagcccgaga acaactacaa gaccaccccc 1200

ccagtgctgg acagcgacgg cagcttcttc ctgtacagca agctgaccgt ggacaagtcc 1260

aggtggcagc agggcaacgt gttcagctgc agcgtgatgc acgaggccct gcacaaccac 1320

tacacccaga agtccctgag cctgagcccc ggcaag 1356

<210> 324

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 324

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 325

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 325

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 326

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 326

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 327

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 327

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 328

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 328

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 329

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 329

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 330

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 330

Ser Gln Ser Ile Ser Asn Asn

fifteen

<210> 331

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 331

Tyr Ala Ser

one

<210> 332

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 332

Ser Ser Ser Trp Leu

fifteen

<210> 333

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 333

Gln Ser Ile Ser Asn Asn

fifteen

<210> 334

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 334

Tyr Ala Ser

one

<210> 335

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 335

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 336

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 336

Asp Ile 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 Ser Ile Ser Asn Asn

20 25 30

Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Lys 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 Ser Val Glu Thr

65 70 75 80

Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ser Asn Ser Trp Leu Thr

85 90 95

Phe Gly Ala Gly Thr Lys Leu Gly Leu Lys

100 105

<210> 337

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 337

gatattgtgc taactcagtc tccagccacc ctgtctgtga ctccaggaga tagcgtcagt 60

ctttcctgca gggccagcca aagtattagc aacaacctac actggtatca acagaaatca 120

catgagtctc caaaacttct catcaagtat gcttcccagt ccatctctgg gatcccctcc 180

aggttcagtg gcagtggatc agggacagat ttcactctca gtatcaacag tgtggagact 240

gaagattttg gaatgtattt ctgtcaacag agtaacagct ggctcacgtt cggtgctggg 300

accaagctggggctgaaa 318

<210> 338

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 338

Asp Ile 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 Ser Ile Ser Asn Asn

20 25 30

Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Lys 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 Ser Val Glu Thr

65 70 75 80

Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ser Asn Ser Trp Leu Thr

85 90 95

Phe Gly Ala Gly Thr Lys Leu Gly Leu Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 339

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 339

gatattgtgc taactcagtc tccagccacc ctgtctgtga ctccaggaga tagcgtcagt 60

ctttcctgca gggccagcca aagtattagc aacaacctac actggtatca acagaaatca 120

catgagtctc caaaacttct catcaagtat gcttcccagt ccatctctgg gatcccctcc 180

aggttcagtg gcagtggatc agggacagat ttcactctca gtatcaacag tgtggagact 240

gaagattttg gaatgtattt ctgtcaacag agtaacagct ggctcacgtt cggtgctggg 300

accaagctgg ggctgaaacg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 340

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 340

Gly Phe Thr Phe Ser Thr Tyr Ala Met Ser

1.5 10

<210> 341

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 341

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 342

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 342

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 343

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 343

Thr Tyr Ala Met Ser

fifteen

<210> 344

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 344

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 345

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 345

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 346

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 346

Gly Phe Thr Phe Ser Thr Tyr

fifteen

<210> 347

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 347

Ser Asp Ala Gly Ser Tyr

fifteen

<210> 348

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 348

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 349

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 349

Gly Phe Thr Phe Ser Thr Tyr Ala

fifteen

<210> 350

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 350

Ile Ser Asp Ala Gly Ser Tyr Ser

fifteen

<210> 351

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 351

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10 15

<210> 352

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 352

Glu Val Gln Leu Val Glu 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 Phe 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 Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Glyn Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 353

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 353

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc acctacgcca tgtcctggggt gcggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctccgacg ccggctccta ctcctactac 180

cccgacaacg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacggggc 300

tccagatacg aagagtacta cgtgatggac tattggggcc agggcaccac cgtgacagtg 360

tcctcc 366

<210> 354

<211> 452

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 354

Glu Val Gln Leu Val Glu 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 Phe 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 Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 355

<211> 1356

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 355

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc acctacgcca tgtcctggggt gcggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctccgacg ccggctccta ctcctactac 180

cccgacaacg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacggggc 300

tccagatacg aagagtacta cgtgatggac tattggggcc agggcaccac cgtgacagtg 360

tcctccgcta gcaccaaggg cccaagtgtg tttcccctgg cccccagcag caagtctact 420

tccggcggaa ctgctgccct gggttgcctg gtgaaggact acttcccctg tcccgtgaca 480

gtgtcctgga actctggggc tctgacttcc ggcgtgcaca ccttccccgc cgtgctgcag 540

agcagcggcc tgtacagcct gagcagcgtg gtgacagtgc cctccagctc tctgggaacc 600

cagacctata tctgcaacgt gaaccacaag cccagcaaca ccaaggtgga caagagagtg 660

gagcccaaga gctgcgacaa gacccacacc tgccccccct gcccagctcc agaactgctg 720

ggagggcctt ccgtgttcct gttccccccc aagcccaagg acaccctgat gatcagcagg 780

acccccgagg tgacctgcgt ggtggtggcc gtgtcccacg aggacccaga ggtgaagttc 840

aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcccag agaggagcag 900

tacaacagca cctacagggt ggtgtccgtg ctgaccgtgc tgcaccagga ctggctgaac 960

ggcaaagaat acaagtgcaa agtctccaac aaggccctgg ctgccccaat cgaaaagaca 1020

atcagcaagg ccaagggcca gccacgggag ccccaggtgt acaccctgcc ccccagccgg 1080

gaggagatga ccaagaacca ggtgtccctg acctgtctgg tgaagggctt ctacccctgt 1140

gatatcgccg tggagtggga gagcaacggc cagcccgaga acaactacaa gaccaccccc 1200

ccagtgctgg acagcgacgg cagcttcttc ctgtacagca agctgaccgt ggacaagtcc 1260

aggtggcagc agggcaacgt gttcagctgc agcgtgatgc acgaggccct gcacaaccac 1320

tacacccaga agtccctgag cctgagcccc ggcaag 1356

<210> 356

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 356

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 357

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 357

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 358

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 358

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 359

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 359

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 360

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 360

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 361

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 361

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 362

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 362

Ser Gln Ser Ile Ser Asn Asn

fifteen

<210> 363

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 363

Tyr Ala Ser

one

<210> 364

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 364

Ser Ser Ser Trp Leu

fifteen

<210> 365

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 365

Gln Ser Ile Ser Asn Asn

fifteen

<210> 366

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 366

Tyr Ala Ser

one

<210> 367

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 367

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 368

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 368

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser 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 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 Val Glu Pro

65 70 75 80

Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Ser Ser Trp Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 369

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 369

gagatcgtgc tgacccagtc ccctgccacc ctgtccgtgt ctcccggcga gagagtgacc 60

ctgtcctgcc gggcctccca gtccatctcc aacaacctgc actggtatca gcagaagccc 120

ggccaggccc ctcggctgct gattaagtac gcctcccaga gcatctccgg catccctgcc 180

agattctccg gctccggcag cggcaccgac ttcaccctga ccatctccag cgtggaaccc 240

gaggacttcg gcgtgtactt ctgccagcag tcctcatcct ggctgacctt cggccaggggc 300

accaagctgg aaatcaag 318

<210> 370

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 370

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser 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 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 Val Glu Pro

65 70 75 80

Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Ser Ser Trp Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 371

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 371

gagatcgtgc tgacccagtc ccctgccacc ctgtccgtgt ctcccggcga gagagtgacc 60

ctgtcctgcc gggcctccca gtccatctcc aacaacctgc actggtatca gcagaagccc 120

ggccaggccc ctcggctgct gattaagtac gcctcccaga gcatctccgg catccctgcc 180

agattctccg gctccggcag cggcaccgac ttcaccctga ccatctccag cgtggaaccc 240

gaggacttcg gcgtgtactt ctgccagcag tcctcatcct ggctgacctt cggccaggggc 300

accaagctgg aaatcaagcg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 372

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 372

Gly Phe Thr Phe Ser Tyr Ala Met Ser

1.5 10

<210> 373

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 373

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 374

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 374

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 375

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 375

Ser Tyr Ala Met Ser

fifteen

<210> 376

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 376

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 377

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 377

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 378

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 378

Gly Phe Thr Phe Ser Ser Tyr

fifteen

<210> 379

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 379

Ser Ser Gly Gly Ser Phe

fifteen

<210> 380

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 380

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 381

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 381

Gly Phe Thr Phe Ser Ser Tyr Ala

fifteen

<210> 382

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 382

Ile Ser Ser Gly Gly Ser Phe Thr

fifteen

<210> 383

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 383

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 384

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 384

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 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Ala Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 385

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 385

gaagtgatgc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt agctatgcca tgtcttggat tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc atcagtagtg gtggtagttt cacctactat 180

ccagacagtg tgaaggggcg attcaccatt tctagagaca atgtcaagaa caccctgtac 240

ctgcaaatga gcagtctgag gtctgaagac acggccatgt attactgtgc aagacgggct 300

tctacggtag taggtacgga cttcgatgtc tggggcgcag ggaccacggt caccgtctcc 360

tca 363

<210> 386

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 386

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 Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Ala Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 387

<211> 1353

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 387

gaagtgatgc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt agctatgcca tgtcttggat tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc atcagtagtg gtggtagttt cacctactat 180

ccagacagtg tgaaggggcg attcaccatt tctagagaca atgtcaagaa caccctgtac 240

ctgcaaatga gcagtctgag gtctgaagac acggccatgt attactgtgc aagacgggct 300

tctacggtag taggtacgga cttcgatgtc tggggcgcag ggaccacggt caccgtctcc 360

tcagctagca ccaagggccc aagtgtgttt cccctggccc ccagcagcaa gtctacttcc 420

ggcggaactg ctgccctggg ttgcctggtg aaggactact tcccctgtcc cgtgacagtg 480

tcctggaact ctggggctct gacttccggc gtgcacacct tccccgccgt gctgcagagc 540

agcggcctgt acagcctgag cagcgtggtg acagtgccct ccagctctct gggaacccag 600

acctatatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gagagtggag 660

cccaagagct gcgacaagac ccacacctgc cccccctgcc cagctccaga actgctggga 720

gggccttccg tgttcctgtt cccccccaag cccaaggaca ccctgatgat cagcaggacc 780

cccgaggtga cctgcgtggt ggtggacgtg tcccacgagg acccagaggt gaagttcaac 840

tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcccagaga ggagcagtac 900

aacagcacct acagggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 960

aaagaataca agtgcaaagt ctccaacaag gccctgccag ccccaatcga aaagacaatc 1020

1080

gagatgacca agaaccaggt gtccctgacc tgtctggtga agggcttcta cccctgtgat 1140

atcgccgtgg agtgggagag caacggccag cccgagaaca actacaagac caccccccca 1200

1260

tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1320

acccagaagt ccctgagcct gagccccggc aag 1353

<210> 388

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 388

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 389

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 389

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 390

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 390

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 391

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 391

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 392

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 392

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 393

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 393

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 394

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 394

Ser Gln Asp Ile Gly Ser Ser

fifteen

<210> 395

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 395

Ala Thr Ser

one

<210> 396

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 396

Tyr Ala Ser Ser Pro

fifteen

<210> 397

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 397

Gln Asp Ile Gly Ser Ser

fifteen

<210> 398

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 398

Ala Thr Ser

one

<210> 399

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 399

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 400

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 400

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly

1.5 10 15

Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 401

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 401

gacatccaga tgacccagtc tccatcctcc ttatctgcct ctctgggaga aagagtcagt 60

ctcacttgtc gggcaagtca ggacattggt agtagcttaa actggcttca gcaggaacca 120

gatggaacta ttaaacgcct gatctatgcc acatccagtt tagattctgg tgtccccaaa 180

aggttcagtg gcagtaggtc tgggtcagat tattctctca ccatcagcag ccttgagtct 240

gaagattttg tagtctatta ctgtctacaa tatgctagtt cgcctccgac gttcggtgga 300

ggcaccaagc tggaaatcaa a 321

<210> 402

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 402

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly

1.5 10 15

Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 403

<211> 642

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 403

gacatccaga tgacccagtc tccatcctcc ttatctgcct ctctgggaga aagagtcagt 60

ctcacttgtc gggcaagtca ggacattggt agtagcttaa actggcttca gcaggaacca 120

gatggaacta ttaaacgcct gatctatgcc acatccagtt tagattctgg tgtccccaaa 180

aggttcagtg gcagtaggtc tgggtcagat tattctctca ccatcagcag ccttgagtct 240

gaagattttg tagtctatta ctgtctacaa tatgctagtt cgcctccgac gttcggtgga 300

ggcaccaagc tggaaatcaa acgtacggtg gccgctccca gcgtgttcat cttccccccc 360

agcgacgagc agctgaagag tggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420

ccccgggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480

gagagcgtca ccgagcagga cagcaaggac tccacctaca gcctgagcag caccctgacc 540

ctgagcaagg ccgactacga gaagcataag gtgtacgcct gcgaggtgac ccaccagggc 600

ctgtccagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642

<210> 404

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 404

Gly Phe Thr Phe Ser Tyr Ala Met Ser

1.5 10

<210> 405

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 405

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 406

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 406

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 407

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 407

Ser Tyr Ala Met Ser

fifteen

<210> 408

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 408

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 409

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 409

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 410

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 410

Gly Phe Thr Phe Ser Ser Tyr

fifteen

<210> 411

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 411

Ser Ser Gly Gly Ser Phe

fifteen

<210> 412

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 412

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 413

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 413

Gly Phe Thr Phe Ser Ser Tyr Ala

fifteen

<210> 414

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 414

Ile Ser Ser Gly Gly Ser Phe Thr

fifteen

<210> 415

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 415

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 416

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 416

Glu Val Gln Leu Val Glu 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 Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 417

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 417

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc agctacgcca tgtcctggat ccggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctcctccg gcggcagctt cacctactac 180

cccgcagcg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacgggcc 300

tccaccgtcg tgggaaccga cttcgatgtg tggggccagg gcaccaccgt gacagtgtcc 360

tcc 363

<210> 418

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 418

Glu Val Gln Leu Val Glu 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 Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 419

<211> 1353

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 419

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc agctacgcca tgtcctggat ccggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctcctccg gcggcagctt cacctactac 180

cccgcagcg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacgggcc 300

tccaccgtcg tgggaaccga cttcgatgtg tggggccagg gcaccaccgt gacagtgtcc 360

tccgctagca ccaagggccc aagtgtgttt cccctggccc ccagcagcaa gtctacttcc 420

ggcggaactg ctgccctggg ttgcctggtg aaggactact tcccctgtcc cgtgacagtg 480

tcctggaact ctggggctct gacttccggc gtgcacacct tccccgccgt gctgcagagc 540

agcggcctgt acagcctgag cagcgtggtg acagtgccct ccagctctct gggaacccag 600

acctatatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gagagtggag 660

cccaagagct gcgacaagac ccacacctgc cccccctgcc cagctccaga actgctggga 720

gggccttccg tgttcctgtt cccccccaag cccaaggaca ccctgatgat cagcaggacc 780

cccgaggtga cctgcgtggt ggtggccgtg tcccacgagg acccagaggt gaagttcaac 840

tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcccagaga ggagcagtac 900

aacagcacct acagggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 960

aaagaataca agtgcaaagt ctccaacaag gccctggctg ccccaatcga aaagacaatc 1020

1080

gagatgacca agaaccaggt gtccctgacc tgtctggtga agggcttcta cccctgtgat 1140

atcgccgtgg agtgggagag caacggccag cccgagaaca actacaagac caccccccca 1200

1260

tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1320

acccagaagt ccctgagcct gagccccggc aag 1353

<210> 420

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 420

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 421

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 421

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 422

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 422

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 423

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 423

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 424

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 424

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 425

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 425

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 426

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 426

Ser Gln Asp Ile Gly Ser Ser

fifteen

<210> 427

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 427

Ala Thr Ser

one

<210> 428

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 428

Tyr Ala Ser Ser Pro

fifteen

<210> 429

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 429

Gln Asp Ile Gly Ser Ser

fifteen

<210> 430

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 430

Ala Thr Ser

one

<210> 431

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 431

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 432

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 432

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1.5 10 15

Asp Arg Val Thr Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Lys Pro Gly Lys Ala Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 433

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 433

gacatccaga tgacccagtc cccctccagc ctgtccgcct ccgtgggcga tagagtgacc 60

ctgacctgcc gggcctccca ggacatcggc tcctccctga actggctgca gcagaagccc 120

ggcaaggcca tcaagcggct gatctacgcc acctcctccc tggactccgg cgtgccctcc 180

cggttctccg gctctagatc cggcaccgac tacaccctga ccatctccag cctgcagccc 240

gaggacttcg tggtgtacta ctgcctgcag tacgcctcca gcccccccac ctttggcgga 300

ggcaccaagc tggaaatcaa g 321

<210> 434

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 434

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1.5 10 15

Asp Arg Val Thr Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Lys Pro Gly Lys Ala Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 435

<211> 642

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 435

gacatccaga tgacccagtc cccctccagc ctgtccgcct ccgtgggcga tagagtgacc 60

ctgacctgcc gggcctccca ggacatcggc tcctccctga actggctgca gcagaagccc 120

ggcaaggcca tcaagcggct gatctacgcc acctcctccc tggactccgg cgtgccctcc 180

cggttctccg gctctagatc cggcaccgac tacaccctga ccatctccag cctgcagccc 240

gaggacttcg tggtgtacta ctgcctgcag tacgcctcca gcccccccac ctttggcgga 300

ggcaccaagc tggaaatcaa gcgtacggtg gccgctccca gcgtgttcat cttccccccc 360

agcgacgagc agctgaagag tggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420

ccccgggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480

gagagcgtca ccgagcagga cagcaaggac tccacctaca gcctgagcag caccctgacc 540

ctgagcaagg ccgactacga gaagcataag gtgtacgcct gcgaggtgac ccaccagggc 600

ctgtccagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642

<210> 436

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 436

Gly Phe Thr Phe Ser Asn Phe Ala Met Ser

1.5 10

<210> 437

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 437

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 438

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 438

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 439

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 439

Ser Asn Phe Ala Met Ser

fifteen

<210> 440

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 440

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 441

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 441

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 442

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 442

Gly Phe Thr Phe Ser Asn Phe

fifteen

<210> 443

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 443

Ser Thr Gly Gly Thr Tyr

fifteen

<210> 444

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 444

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 445

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 445

Gly Phe Thr Phe Ser Asn Phe Ala

fifteen

<210> 446

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 446

Ile Ser Thr Gly Gly Thr Tyr Thr

fifteen

<210> 447

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 447

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys

1.5 10

<210> 448

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 448

Glu Val His 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 Asn Phe

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 Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser

115

<210> 449

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 449

gaagtgcatc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt aactttgcca tgtcttgggt tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc attagtactg gtggtactta cacctactat 180

ccagacagtg tgaagggtcg attcaccatc tccagagaca atgccaagaa aaccctgtac 240

ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtac aagacggggg 300

tacgacggcg tggacaaatg gggccaaggc accactctca cagtctcctc a 351

<210> 450

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 450

Glu Val His 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 Asn Phe

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 Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 451

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 451

gaagtgcatc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt aactttgcca tgtcttgggt tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc attagtactg gtggtactta cacctactat 180

ccagacagtg tgaagggtcg attcaccatc tccagagaca atgccaagaa aaccctgtac 240

ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtac aagacggggg 300

tacgacggcg tggacaaatg gggccaaggc accactctca cagtctcctc agctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc ccctgtcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggacgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctgccagcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc cctgtgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 452

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 452

Lys Ser Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 453

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 453

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 454

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 454

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 455

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 455

Lys Ser Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 456

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 456

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 457

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 457

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 458

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 458

Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr

1.5 10

<210> 459

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 459

Leu Val Ser

one

<210> 460

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 460

Gly Thr His Phe Pro Gln

fifteen

<210> 461

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 461

Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr

1.5 10

<210> 462

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 462

Leu Val Ser

one

<210> 463

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 463

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 464

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 464

Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 465

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 465

gatgttgtga tgacccagac tccactcact ttgtcggtta ccattggaca accagcctcc 60

atctcttgca agtcaggtca gagcctctta gatagtgatg gaaagacata tttgaattgg 120

tttttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttcct 300

cagacgttcg gtggaggcac caagctggaa atcaaa 336

<210> 466

<211> 219

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 466

Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 467

<211> 657

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 467

gatgttgtga tgacccagac tccactcact ttgtcggtta ccattggaca accagcctcc 60

atctcttgca agtcaggtca gagcctctta gatagtgatg gaaagacata tttgaattgg 120

tttttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttcct 300

cagacgttcg gtggaggcac caagctggaa atcaaacgta cggtggccgc tcccagcgtg 360

ttcatcttcc cccccagcga cgagcagctg aagagtggca ccgccagcgt ggtgtgcctg 420

ctgaacaact tctacccccg ggaggccaag gtgcagtgga aggtggacaa cgccctgcag 480

agcggcaaca gccaggagag cgtcaccgag caggacagca aggactccac ctacagcctg 540

agcagcaccc tgaccctgag caaggccgac tacgagaagc ataaggtgta cgcctgcgag 600

gtgacccacc agggcctgtc cagccccgtg accaagagct tcaacagggg cgagtgc 657

<210> 468

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 468

Gly Phe Thr Phe Ser Asn Phe Ala Met Ser

1.5 10

<210> 469

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 469

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 470

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 470

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 471

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 471

Ser Asn Phe Ala Met Ser

fifteen

<210> 472

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 472

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 473

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 473

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 474

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 474

Gly Phe Thr Phe Ser Asn Phe

fifteen

<210> 475

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 475

Ser Thr Gly Gly Thr Tyr

fifteen

<210> 476

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 476

Arg Gly Tyr Asp Gly Val Asp Lys

fifteen

<210> 477

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 477

Gly Phe Thr Phe Ser Asn Phe Ala

fifteen

<210> 478

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 478

Ile Ser Thr Gly Gly Thr Tyr Thr

fifteen

<210> 479

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 479

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys

1.5 10

<210> 480

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 480

Glu Val His 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 Asn Phe

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 Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser

115

<210> 481

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 481

gaagtgcatc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt aactttgcca tgtcttgggt tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc attagtactg gtggtactta cacctactat 180

ccagacagtg tgaagggtcg attcaccatc tccagagaca atgccaagaa aaccctgtac 240

ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtac aagacggggg 300

tacgacggcg tggacaaatg gggccaaggc accactctca cagtctcctc a 351

<210> 482

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 482

Glu Val His 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 Asn Phe

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 Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Thr Leu Tyr

65 70 75 80

Leu Gln Met Ser Ser Leu Arg Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Thr Arg Arg Gly Tyr Asp Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 483

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 483

gaagtgcatc tggtggagtc tgggggaggc ttagtgaagc ctggagggtc cctgaaactc 60

tcctgtgcag cctctggatt cactttcagt aactttgcca tgtcttgggt tcgccagact 120

ccggagaaga gactggagtg ggtcgcaacc attagtactg gtggtactta cacctactat 180

ccagacagtg tgaagggtcg attcaccatc tccagagaca atgccaagaa aaccctgtac 240

ctgcaaatga gcagtctgag gtctgaggac acggccatgt attactgtac aagacggggg 300

tacgacggcg tggacaaatg gggccaaggc accactctca cagtctcctc agctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc ccctgtcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc cctgtgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 484

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 484

Lys Ser Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 485

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 485

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 486

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 486

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 487

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 487

Lys Ser Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 488

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 488

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 489

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 489

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 490

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 490

Gly Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr

1.5 10

<210> 491

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 491

Leu Val Ser

one

<210> 492

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 492

Gly Thr His Phe Pro Gln

fifteen

<210> 493

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 493

Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr

1.5 10

<210> 494

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 494

Leu Val Ser

one

<210> 495

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 495

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 496

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 496

Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 497

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 497

gatgttgtga tgacccagac tccactcact ttgtcggtta ccattggaca accagcctcc 60

atctcttgca agtcaggtca gagcctctta gatagtgatg gaaagacata tttgaattgg 120

tttttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttcct 300

cagacgttcg gtggaggcac caagctggaa atcaaa 336

<210> 498

<211> 219

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 498

Asp Val Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val Thr Ile Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Asp Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 499

<211> 657

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 499

gatgttgtga tgacccagac tccactcact ttgtcggtta ccattggaca accagcctcc 60

atctcttgca agtcaggtca gagcctctta gatagtgatg gaaagacata tttgaattgg 120

tttttacaga ggccaggcca gtctccaaag cgcctaatct atctggtgtc taaactggac 180

tctggagtcc ctgacaggtt cactggcagt ggatcaggga cagatttcac actgaaaatc 240

agcagagtgg aggctgagga tttgggagtt tattattgct ggcaaggtac acattttcct 300

cagacgttcg gtggaggcac caagctggaa atcaaacgta cggtggccgc tcccagcgtg 360

ttcatcttcc cccccagcga cgagcagctg aagagtggca ccgccagcgt ggtgtgcctg 420

ctgaacaact tctacccccg ggaggccaag gtgcagtgga aggtggacaa cgccctgcag 480

agcggcaaca gccaggagag cgtcaccgag caggacagca aggactccac ctacagcctg 540

agcagcaccc tgaccctgag caaggccgac tacgagaagc ataaggtgta cgcctgcgag 600

gtgacccacc agggcctgtc cagccccgtg accaagagct tcaacagggg cgagtgc 657

<210> 500

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 500

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser Trp His

1.5 10

<210> 501

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 501

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 502

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 502

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 503

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 503

Ser Gly Tyr Ser Trp His

fifteen

<210> 504

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 504

His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1.5 10 15

<210> 505

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 505

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 506

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 506

Gly Tyr Ser Ile Thr Ser Gly Tyr

fifteen

<210> 507

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 507

His Ser Ser Gly Ser

fifteen

<210> 508

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 508

Gly Gly Val Gln Ala Phe Ala Tyr

fifteen

<210> 509

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 509

Gly Tyr Ser Ile Thr Ser Gly Tyr Ser

fifteen

<210> 510

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 510

Ile His Ser Ser Gly Ser Thr

fifteen

<210> 511

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 511

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr

1.5 10

<210> 512

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 512

Asp 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 Val Ser Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 513

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 513

gacgtgcagc tgcaggaatc tggccctggc ctggtgaaac cctcccagac cctgtccctg 60

acctgcaccg tgtccggcta ctccatcacc tccggctaca gctggcactg gatccggcag 120

caccccggca agggcctgga atggatggcc cacatccact cctccggctc caccaactac 180

aaccccagcc tgaagtccag aatcaccatc agccgggaca cctccaagaa ccagttctcc 240

ctgaagctgt cctccgtgac cgccgctgac accgccgtgt actactgtgc cagaggcggc 300

gtgcaggcct tcgcttattg gggccagggc accctggtga cagtgtcctc c 351

<210> 514

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 514

Asp 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 Val Ser Gly Tyr Ser Ile Thr Ser Gly

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu Trp

35 40 45

Met Ala His Ile His Ser Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Val Gln Ala Phe Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 515

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 515

gacgtgcagc tgcaggaatc tggccctggc ctggtgaaac cctcccagac cctgtccctg 60

acctgcaccg tgtccggcta ctccatcacc tccggctaca gctggcactg gatccggcag 120

caccccggca agggcctgga atggatggcc cacatccact cctccggctc caccaactac 180

aaccccagcc tgaagtccag aatcaccatc agccgggaca cctccaagaa ccagttctcc 240

ctgaagctgt cctccgtgac cgccgctgac accgccgtgt actactgtgc cagaggcggc 300

gtgcaggcct tcgcttattg gggccagggc accctggtga cagtgtcctc cgctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc cccgagcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 516

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 516

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 517

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 517

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 518

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 518

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 519

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 519

Ser Ala Ser Ser Ser Val Ile Tyr Met His

1.5 10

<210> 520

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 520

Asp Thr Ser Lys Leu Ala Ser

fifteen

<210> 521

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 521

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 522

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 522

Ser Ser Ser Val Ile Tyr

fifteen

<210> 523

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 523

Asp Thr Ser

one

<210> 524

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 524

Trp Ser Ser Asn Pro Leu

fifteen

<210> 525

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 525

Ser Ser Val Ile Tyr

fifteen

<210> 526

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 526

Asp Thr Ser

one

<210> 527

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 527

Gln Gln Trp Ser Ser Asn Pro Leu Thr

fifteen

<210> 528

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 528

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Ala Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Met Ser Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Met Glu Pro Glu

65 70 75 80

Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 529

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 529

gagatcgtgc tgacccagtc ccctgccacc ctgtctgcta gccctggcga gcgcgtgaca 60

atgtcctgct ccgcctcctc ctccgtgatc tacatgcact ggtatcagca gaagcccggc 120

caggcccctc ggcggtggat ctacgatacc tccaagctgg cctccggcgt gcccgccaga 180

ttctccggct ctggctctgg caccgactac accctgacca tctccagcat ggaacccgag 240

gacgccgccg tgtactactg cggcagtgg tcctccaacc ccctgacctt cggccaggggc 300

accaagctgg aaatcaag 318

<210> 530

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 530

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Ala Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Met Ser Cys Ser Ala Ser Ser Ser Val Ile Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Met Glu Pro Glu

65 70 75 80

Asp Ala Ala Val Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 531

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 531

gagatcgtgc tgacccagtc ccctgccacc ctgtctgcta gccctggcga gcgcgtgaca 60

atgtcctgct ccgcctcctc ctccgtgatc tacatgcact ggtatcagca gaagcccggc 120

caggcccctc ggcggtggat ctacgatacc tccaagctgg cctccggcgt gcccgccaga 180

ttctccggct ctggctctgg caccgactac accctgacca tctccagcat ggaacccgag 240

gacgccgccg tgtactactg cggcagtgg tcctccaacc ccctgacctt cggccaggggc 300

accaagctgg aaatcaagcg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 532

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 532

Gly Phe Thr Phe Ser Thr Tyr Ala Met Ser

1.5 10

<210> 533

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 533

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 534

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 534

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 535

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 535

Thr Tyr Ala Met Ser

fifteen

<210> 536

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 536

Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val Lys

1.5 10 15

gly

<210> 537

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 537

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 538

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 538

Gly Phe Thr Phe Ser Thr Tyr

fifteen

<210> 539

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 539

Ser Asp Ala Gly Ser Tyr

fifteen

<210> 540

<211> 13

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 540

Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10

<210> 541

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 541

Gly Phe Thr Phe Ser Thr Tyr Ala

fifteen

<210> 542

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 542

Ile Ser Asp Ala Gly Ser Tyr Ser

fifteen

<210> 543

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 543

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr

1.5 10 15

<210> 544

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 544

Glu Val Gln Leu Val Glu 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 Phe 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 Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 545

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 545

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc acctacgcca tgtcctggggt gcggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctccgacg ccggctccta ctcctactac 180

cccgacaacg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacggggc 300

tccagatacg aagagtacta cgtgatggac tattggggcc agggcaccac cgtgacagtg 360

tcctcc 366

<210> 546

<211> 452

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 546

Glu Val Gln Leu Val Glu 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 Phe 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 Thr Ile Ser Asp Ala Gly Ser Tyr Ser Tyr Tyr Pro Asp Asn Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ser Arg Tyr Glu Glu Tyr Tyr Val Met Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 547

<211> 1356

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 547

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc acctacgcca tgtcctggggt gcggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctccgacg ccggctccta ctcctactac 180

cccgacaacg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacggggc 300

tccagatacg aagagtacta cgtgatggac tattggggcc agggcaccac cgtgacagtg 360

tcctccgcta gcaccaaggg cccaagtgtg tttcccctgg cccccagcag caagtctact 420

tccggcggaa ctgctgccct gggttgcctg gtgaaggact acttccccga gcccgtgaca 480

gtgtcctgga actctggggc tctgacttcc ggcgtgcaca ccttccccgc cgtgctgcag 540

agcagcggcc tgtacagcct gagcagcgtg gtgacagtgc cctccagctc tctgggaacc 600

cagacctata tctgcaacgt gaaccacaag cccagcaaca ccaaggtgga caagagagtg 660

gagcccaaga gctgcgacaa gacccacacc tgccccccct gcccagctcc agaactgctg 720

ggagggcctt ccgtgttcct gttccccccc aagcccaagg acaccctgat gatcagcagg 780

acccccgagg tgacctgcgt ggtggtggcc gtgtcccacg aggacccaga ggtgaagttc 840

aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcccag agaggagcag 900

tacaacagca cctacagggt ggtgtccgtg ctgaccgtgc tgcaccagga ctggctgaac 960

ggcaaagaat acaagtgcaa agtctccaac aaggccctgg ctgccccaat cgaaaagaca 1020

atcagcaagg ccaagggcca gccacgggag ccccaggtgt acaccctgcc ccccagccgg 1080

gaggagatga ccaagaacca ggtgtccctg acctgtctgg tgaagggctt ctaccccagc 1140

gatatcgccg tggagtggga gagcaacggc cagcccgaga acaactacaa gaccaccccc 1200

ccagtgctgg acagcgacgg cagcttcttc ctgtacagca agctgaccgt ggacaagtcc 1260

aggtggcagc agggcaacgt gttcagctgc agcgtgatgc acgaggccct gcacaaccac 1320

tacacccaga agtccctgag cctgagcccc ggcaag 1356

<210> 548

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 548

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 549

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 549

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 550

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 550

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 551

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 551

Arg Ala Ser Gln Ser Ile Ser Asn Asn Leu His

1.5 10

<210> 552

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 552

Tyr Ala Ser Gln Ser Ile Ser

fifteen

<210> 553

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 553

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 554

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 554

Ser Gln Ser Ile Ser Asn Asn

fifteen

<210> 555

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 555

Tyr Ala Ser

one

<210> 556

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 556

Ser Ser Ser Trp Leu

fifteen

<210> 557

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 557

Gln Ser Ile Ser Asn Asn

fifteen

<210> 558

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 558

Tyr Ala Ser

one

<210> 559

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 559

Gln Gln Ser Ser Ser Trp Leu Thr

fifteen

<210> 560

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 560

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser 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 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 Val Glu Pro

65 70 75 80

Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Ser Ser Trp Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 561

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 561

gagatcgtgc tgacccagtc ccctgccacc ctgtccgtgt ctcccggcga gagagtgacc 60

ctgtcctgcc gggcctccca gtccatctcc aacaacctgc actggtatca gcagaagccc 120

ggccaggccc ctcggctgct gattaagtac gcctcccaga gcatctccgg catccctgcc 180

agattctccg gctccggcag cggcaccgac ttcaccctga ccatctccag cgtggaaccc 240

gaggacttcg gcgtgtactt ctgccagcag tcctcatcct ggctgacctt cggccaggggc 300

accaagctgg aaatcaag 318

<210> 562

<211> 213

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 562

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1.5 10 15

Glu Arg Val Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser 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 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 Val Glu Pro

65 70 75 80

Glu Asp Phe Gly Val Tyr Phe Cys Gln Gln Ser Ser Ser Trp Leu Thr

85 90 95

Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu Cys

210

<210> 563

<211> 639

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 563

gagatcgtgc tgacccagtc ccctgccacc ctgtccgtgt ctcccggcga gagagtgacc 60

ctgtcctgcc gggcctccca gtccatctcc aacaacctgc actggtatca gcagaagccc 120

ggccaggccc ctcggctgct gattaagtac gcctcccaga gcatctccgg catccctgcc 180

agattctccg gctccggcag cggcaccgac ttcaccctga ccatctccag cgtggaaccc 240

gaggacttcg gcgtgtactt ctgccagcag tcctcatcct ggctgacctt cggccaggggc 300

accaagctgg aaatcaagcg tacggtggcc gctcccagcg tgttcatctt cccccccagc 360

gacgagcagc tgaagagtgg caccgccagc gtggtgtgcc tgctgaacaa cttctacccc 420

cgggaggcca aggtgcagtg gaaggtggac aacgccctgc agagcggcaa cagccaggag 480

agcgtcaccg agcaggacag caaggactcc acctacagcc tgagcagcac cctgaccctg 540

agcaaggccg actacgagaa gcataaggtg tacgcctgcg aggtgaccca ccagggcctg 600

tccagccccg tgaccaagag cttcaacagg ggcgagtgc 639

<210> 564

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 564

Gly Phe Thr Phe Ser Tyr Ala Met Ser

1.5 10

<210> 565

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 565

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 566

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 566

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 567

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 567

Ser Tyr Ala Met Ser

fifteen

<210> 568

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 568

Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 569

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 569

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 570

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 570

Gly Phe Thr Phe Ser Ser Tyr

fifteen

<210> 571

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 571

Ser Ser Gly Gly Ser Phe

fifteen

<210> 572

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 572

Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 573

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 573

Gly Phe Thr Phe Ser Ser Tyr Ala

fifteen

<210> 574

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 574

Ile Ser Ser Gly Gly Ser Phe Thr

fifteen

<210> 575

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 575

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val

1.5 10

<210> 576

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 576

Glu Val Gln Leu Val Glu 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 Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 577

<211> 363

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 577

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc agctacgcca tgtcctggat ccggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctcctccg gcggcagctt cacctactac 180

cccgcagcg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacgggcc 300

tccaccgtcg tgggaaccga cttcgatgtg tggggccagg gcaccaccgt gacagtgtcc 360

tcc 363

<210> 578

<211> 451

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 578

Glu Val Gln Leu Val Glu 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 Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Ser Gly Gly Ser Phe Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Ala Ser Thr Val Val Gly Thr Asp Phe Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 579

<211> 1353

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 579

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc agctacgcca tgtcctggat ccggcaggct 120

cccggcaagg gcctggaatg ggtggccacc atctcctccg gcggcagctt cacctactac 180

cccgcagcg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacgggcc 300

tccaccgtcg tgggaaccga cttcgatgtg tggggccagg gcaccaccgt gacagtgtcc 360

tccgctagca ccaagggccc aagtgtgttt cccctggccc ccagcagcaa gtctacttcc 420

ggcggaactg ctgccctgggg ttgcctggtg aaggactact tccccgagcc cgtgacagtg 480

tcctggaact ctggggctct gacttccggc gtgcacacct tccccgccgt gctgcagagc 540

agcggcctgt acagcctgag cagcgtggtg acagtgccct ccagctctct gggaacccag 600

acctatatct gcaacgtgaa ccacaagccc agcaacacca aggtggacaa gagagtggag 660

cccaagagct gcgacaagac ccacacctgc cccccctgcc cagctccaga actgctggga 720

gggccttccg tgttcctgtt cccccccaag cccaaggaca ccctgatgat cagcaggacc 780

cccgaggtga cctgcgtggt ggtggccgtg tcccacgagg acccagaggt gaagttcaac 840

tggtacgtgg acggcgtgga ggtgcacaac gccaagacca agcccagaga ggagcagtac 900

aacagcacct acagggtggt gtccgtgctg accgtgctgc accaggactg gctgaacggc 960

aaagaataca agtgcaaagt ctccaacaag gccctggctg ccccaatcga aaagacaatc 1020

1080

gagatgacca agaaccaggt gtccctgacc tgtctggtga agggcttcta ccccagcgat 1140

atcgccgtgg agtgggagag caacggccag cccgagaaca actacaagac caccccccca 1200

1260

tggcagcagg gcaacgtgtt cagctgcagc gtgatgcacg aggccctgca caaccactac 1320

acccagaagt ccctgagcct gagccccggc aag 1353

<210> 580

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 580

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 581

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 581

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 582

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 582

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 583

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 583

Arg Ala Ser Gln Asp Ile Gly Ser Ser Leu Asn

1.5 10

<210> 584

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 584

Ala Thr Ser Ser Leu Asp Ser

fifteen

<210> 585

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 585

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 586

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 586

Ser Gln Asp Ile Gly Ser Ser

fifteen

<210> 587

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 587

Ala Thr Ser

one

<210> 588

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 588

Tyr Ala Ser Ser Pro

fifteen

<210> 589

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 589

Gln Asp Ile Gly Ser Ser

fifteen

<210> 590

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 590

Ala Thr Ser

one

<210> 591

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 591

Leu Gln Tyr Ala Ser Ser Pro Pro Thr

fifteen

<210> 592

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 592

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1.5 10 15

Asp Arg Val Thr Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Lys Pro Gly Lys Ala Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 593

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 593

gacatccaga tgacccagtc cccctccagc ctgtccgcct ccgtgggcga tagagtgacc 60

ctgacctgcc gggcctccca ggacatcggc tcctccctga actggctgca gcagaagccc 120

ggcaaggcca tcaagcggct gatctacgcc acctcctccc tggactccgg cgtgccctcc 180

cggttctccg gctctagatc cggcaccgac tacaccctga ccatctccag cctgcagccc 240

gaggacttcg tggtgtacta ctgcctgcag tacgcctcca gcccccccac ctttggcgga 300

ggcaccaagc tggaaatcaa g 321

<210> 594

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 594

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1.5 10 15

Asp Arg Val Thr Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Ser Ser

20 25 30

Leu Asn Trp Leu Gln Gln Lys Pro Gly Lys Ala Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr Tyr Cys Leu Gln Tyr Ala Ser Ser Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 595

<211> 642

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 595

gacatccaga tgacccagtc cccctccagc ctgtccgcct ccgtgggcga tagagtgacc 60

ctgacctgcc gggcctccca ggacatcggc tcctccctga actggctgca gcagaagccc 120

ggcaaggcca tcaagcggct gatctacgcc acctcctccc tggactccgg cgtgccctcc 180

cggttctccg gctctagatc cggcaccgac tacaccctga ccatctccag cctgcagccc 240

gaggacttcg tggtgtacta ctgcctgcag tacgcctcca gcccccccac ctttggcgga 300

ggcaccaagc tggaaatcaa gcgtacggtg gccgctccca gcgtgttcat cttccccccc 360

agcgacgagc agctgaagag tggcaccgcc agcgtggtgt gcctgctgaa caacttctac 420

ccccgggagg ccaaggtgca gtggaaggtg gacaacgccc tgcagagcgg caacagccag 480

gagagcgtca ccgagcagga cagcaaggac tccacctaca gcctgagcag caccctgacc 540

ctgagcaagg ccgactacga gaagcataag gtgtacgcct gcgaggtgac ccaccagggc 600

ctgtccagcc ccgtgaccaa gagcttcaac aggggcgagt gc 642

<210> 596

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 596

Gly Phe Thr Phe Ser Asn Phe Ala Met Ser

1.5 10

<210> 597

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 597

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 598

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 598

Arg Gly Tyr Ser Gly Val Asp Lys

fifteen

<210> 599

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 599

Ser Asn Phe Ala Met Ser

fifteen

<210> 600

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 600

Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys

1.5 10 15

gly

<210> 601

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 601

Arg Gly Tyr Ser Gly Val Asp Lys

fifteen

<210> 602

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 602

Gly Phe Thr Phe Ser Asn Phe

fifteen

<210> 603

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 603

Ser Thr Gly Gly Thr Tyr

fifteen

<210> 604

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 604

Arg Gly Tyr Ser Gly Val Asp Lys

fifteen

<210> 605

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 605

Gly Phe Thr Phe Ser Asn Phe Ala

fifteen

<210> 606

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 606

Ile Ser Thr Gly Gly Thr Tyr Thr

fifteen

<210> 607

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 607

Thr Arg Arg Gly Tyr Ser Gly Val Asp Lys

1.5 10

<210> 608

<211> 117

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 608

Glu Val Gln Leu Val Glu 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 Phe Ser Asn Phe

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Tyr Ser Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 609

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 609

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc aacttcgcca tgtcctgggt gcggcaggct 120

cccggcaagg gcctggaatg ggtgtccacc atctccaccg gcggcaccta cacctactac 180

cccgcagcg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacggggc 300

tactcaggcg tggacaaatg gggccagggc accaccgtga cagtgtcctc c 351

<210> 610

<211> 447

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 610

Glu Val Gln Leu Val Glu 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 Phe Ser Asn Phe

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Thr Ile Ser Thr Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Tyr Ser Gly Val Asp Lys Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Cys Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu

260 265 270

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Cys Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 611

<211> 1341

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 611

gaggtgcagc tggtggaatc tggcgggaggc ctggtgaaac ccggcggatc cctgagactg 60

tcctgcgccg cctccggctt caccttctcc aacttcgcca tgtcctgggt gcggcaggct 120

cccggcaagg gcctggaatg ggtgtccacc atctccaccg gcggcaccta cacctactac 180

cccgcagcg tgaagggcag attcaccatc agccgggaca acgccaagaa ctccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtgc cagacggggc 300

tactcaggcg tggacaaatg gggccagggc accaccgtga cagtgtcctc cgctagcacc 360

aagggcccaa gtgtgtttcc cctggccccc agcagcaagt ctacttccgg cggaactgct 420

gccctgggtt gcctggtgaa ggactacttc ccctgtcccg tgacagtgtc ctggaactct 480

ggggctctga cttccggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac agtgccctcc agctctctgg gaacccagac ctatatctgc 600

aacgtgaacc acaagcccag caacaccaag gtggacaaga gagtggagcc caagagctgc 660

gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg 720

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc 780

tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac 840

ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac 900

agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag 960

tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag 1020

ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag 1080

aaccaggtgt ccctgacctg tctggtgaag ggcttctacc cctgtgatat cgccgtggag 1140

tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc 1200

gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcaggggc 1260

aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc 1320

ctgagcctga gccccggcaa g 1341

<210> 612

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 612

Lys Ser Gly Gln Ser Leu Leu Asp Ser Thr Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 613

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 613

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 614

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 614

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 615

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 615

Lys Ser Gly Gln Ser Leu Leu Asp Ser Thr Gly Lys Thr Tyr Leu Asn

1.5 10 15

<210> 616

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 616

Leu Val Ser Lys Leu Asp Ser

fifteen

<210> 617

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 617

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 618

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 618

Gly Gln Ser Leu Leu Asp Ser Thr Gly Lys Thr Tyr

1.5 10

<210> 619

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 619

Leu Val Ser

one

<210> 620

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 620

Gly Thr His Phe Pro Gln

fifteen

<210> 621

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 621

Gln Ser Leu Leu Asp Ser Thr Gly Lys Thr Tyr

1.5 10

<210> 622

<211> 3

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 622

Leu Val Ser

one

<210> 623

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 623

Trp Gln Gly Thr His Phe Pro Gln Thr

fifteen

<210> 624

<211> 112

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 624

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Thr Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Arg Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 625

<211> 336

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 625

gacgtggtga tgacccagtc ccccctgtcc ctgcctgtga ccctgggcca gcctgcctcc 60

atctcctgca agtccggcca gtccctgctg gactccactg gcaagaccta cctgaactgg 120

ttcctgcagc ggcctggcca gtcccctcgg cggctgatct acctggtgtc caagctggac 180

agcggcgtgc ccgacagatt ctccggctct ggctccggca ccgacttcac cctgaagatc 240

tcccgggtgg aagccgagga cgtgggcgtg tactactgct ggcagggcac ccacttcccc 300

cagaccttcg gcggaggcac caagctggaa atcaag 336

<210> 626

<211> 219

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polypeptide"

<400> 626

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1.5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Gly Gln Ser Leu Leu Asp Ser

20 25 30

Thr Gly Lys Thr Tyr Leu Asn Trp Phe Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Arg Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Trp Gln Gly

85 90 95

Thr His Phe Pro Gln Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 627

<211> 657

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

polynucleotide"

<400> 627

gacgtggtga tgacccagtc ccccctgtcc ctgcctgtga ccctgggcca gcctgcctcc 60

atctcctgca agtccggcca gtccctgctg gactccactg gcaagaccta cctgaactgg 120

ttcctgcagc ggcctggcca gtcccctcgg cggctgatct acctggtgtc caagctggac 180

agcggcgtgc ccgacagatt ctccggctct ggctccggca ccgacttcac cctgaagatc 240

tcccgggtgg aagccgagga cgtgggcgtg tactactgct ggcagggcac ccacttcccc 300

cagaccttcg gcggaggcac caagctggaa atcaagcgta cggtggccgc tcccagcgtg 360

ttcatcttcc cccccagcga cgagcagctg aagagtggca ccgccagcgt ggtgtgcctg 420

ctgaacaact tctacccccg ggaggccaag gtgcagtgga aggtggacaa cgccctgcag 480

agcggcaaca gccaggagag cgtcaccgag caggacagca aggactccac ctacagcctg 540

agcagcaccc tgaccctgag caaggccgac tacgagaagc ataaggtgta cgcctgcgag 600

gtgacccacc agggcctgtc cagccccgtg accaagagct tcaacagggg cgagtgc 657

<210> 628

<211> 6

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: Synthetic

6xHis label"

<400> 628

His His His His His His His

fifteen

<210> 629

<211> 22

<212> PROTEIN

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial sequence description: synthetic

peptide"

<400> 629

Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Leu Tyr Arg Ser Pro

1.5 10 15

Ala Met Pro Glu Asn Leu

twenty

<---

Claims (100)

1. An antibody or antigen-binding fragment thereof that binds to the human CCR7 protein, wherein the antibody or antigen-binding fragment thereof has reduced or negligible effector function compared to a wild-type antibody of the same isotype, containing: a. a heavy chain variable region that contains HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO:1, HCDR2 (heavy chain complementarity determining region 2) of SEQ ID NO:2, and HCDR3 (heavy chain complementarity determining region 3) of SEQ ID NO:3; and a light chain variable region which contains LCDR1 (light chain complementarity determining region 1) of SEQ ID NO:17, LCDR2 (light chain complementarity determining region 2) of SEQ ID NO:18, and LCDR3 (light chain complementarity determining region 3) of SEQ ID NO:19; b. a heavy chain variable region that contains HCDR1 of SEQ ID NO:4, HCDR2 of SEQ ID NO:5, and HCDR3 of SEQ ID NO:6; and a light chain variable region that contains LCDR1 of SEQ ID NO:20, LCDR2 of SEQ ID NO:21, and LCDR3 of SEQ ID NO:22; c. a heavy chain variable region that contains HCDR1 of SEQ ID NO:7, HCDR2 of SEQ ID NO:8, and HCDR3 of SEQ ID NO:9; and a light chain variable region that contains LCDR1 of SEQ ID NO:23, LCDR2 of SEQ ID NO:24, and LCDR3 of SEQ ID NO:25; d. a heavy chain variable region that contains HCDR1 of SEQ ID NO:10, HCDR2 of SEQ ID NO:11, and HCDR3 of SEQ ID NO:12; and a light chain variable region that contains LCDR1 of SEQ ID NO:26, LCDR2 of SEQ ID NO:27, and LCDR3 of SEQ ID NO:28; e. a heavy chain variable region that contains HCDR1 of SEQ ID NO:33, HCDR2 of SEQ ID NO:34, and HCDR3 of SEQ ID NO:35; and a light chain variable region that contains LCDR1 of SEQ ID NO:49, LCDR2 of SEQ ID NO:50, and LCDR3 of SEQ ID NO:51; f. a heavy chain variable region that contains HCDR1 of SEQ ID NO:36, HCDR2 of SEQ ID NO:37, and HCDR3 of SEQ ID NO:38; and a light chain variable region that contains LCDR1 of SEQ ID NO:52, LCDR2 of SEQ ID NO:53, and LCDR3 of SEQ ID NO:54; g. a heavy chain variable region that contains HCDR1 of SEQ ID NO:39, HCDR2 of SEQ ID NO:40, and HCDR3 of SEQ ID NO:41; and a light chain variable region that contains LCDR1 of SEQ ID NO:55, LCDR2 of SEQ ID NO:56, and LCDR3 of SEQ ID NO:57; h. a heavy chain variable region that contains HCDR1 of SEQ ID NO:42, HCDR2 of SEQ ID NO:43, and HCDR3 of SEQ ID NO:44; and a light chain variable region that contains LCDR1 of SEQ ID NO:58, LCDR2 of SEQ ID NO:59, and LCDR3 of SEQ ID NO:60; i. a heavy chain variable region that contains HCDR1 of SEQ ID NO:65, HCDR2 of SEQ ID NO:66, and HCDR3 of SEQ ID NO:67; and a light chain variable region that contains LCDR1 of SEQ ID NO:81, LCDR2 of SEQ ID NO:82, and LCDR3 of SEQ ID NO:83; j. a heavy chain variable region that contains HCDR1 of SEQ ID NO:68, HCDR2 of SEQ ID NO:69, and HCDR3 of SEQ ID NO:70; and a light chain variable region that contains LCDR1 of SEQ ID NO:84, LCDR2 of SEQ ID NO:85, and LCDR3 of SEQ ID NO:86; k. a heavy chain variable region that contains HCDR1 of SEQ ID NO:71, HCDR2 of SEQ ID NO:72, and HCDR3 of SEQ ID NO:73; and a light chain variable region that contains LCDR1 of SEQ ID NO:87, LCDR2 of SEQ ID NO:88, and LCDR3 of SEQ ID NO:89; l. a heavy chain variable region that contains HCDR1 of SEQ ID NO:74, HCDR2 of SEQ ID NO:75, and HCDR3 of SEQ ID NO:76; and a light chain variable region that contains LCDR1 of SEQ ID NO:90, LCDR2 of SEQ ID NO:91, and LCDR3 of SEQ ID NO:92; m. a heavy chain variable region that contains HCDR1 of SEQ ID NO:596, HCDR2 of SEQ ID NO:597, and HCDR3 of SEQ ID NO:598; and a light chain variable region that contains LCDR1 of SEQ ID NO:612, LCDR2 of SEQ ID NO:613, and LCDR3 of SEQ ID NO:614; n. a heavy chain variable region that contains HCDR1 of SEQ ID NO:599, HCDR2 of SEQ ID NO:600, and HCDR3 of SEQ ID NO:601; and a light chain variable region that contains LCDR1 of SEQ ID NO:615, LCDR2 of SEQ ID NO:616, and LCDR3 of SEQ ID NO:617; o. a heavy chain variable region that contains HCDR1 of SEQ ID NO:602, HCDR2 of SEQ ID NO:603, and HCDR3 of SEQ ID NO:604; and a light chain variable region that contains LCDR1 of SEQ ID NO:618, LCDR2 of SEQ ID NO:619, and LCDR3 of SEQ ID NO:620; or p. a heavy chain variable region that contains HCDR1 of SEQ ID NO:605, HCDR2 of SEQ ID NO:606, and HCDR3 of SEQ ID NO:607; and a light chain variable region that contains LCDR1 of SEQ ID NO:621, LCDR2 of SEQ ID NO:622, and LCDR3 of SEQ ID NO:623. 2. The antibody of claim. 1, where the antibody or its antigennegative fragment is characterized by a reduced level or negligible level of activity of antibody-dependent cell-mediated cytotoxicity (ADCC). 3. An antibody according to claim 1 or 2, wherein the antibody or antigen-binding fragment thereof contains a silent Fc region. 4. The antibody according to claim 3, where the antibody or its antigen-binding fragment contains a mutation in the Fc region selected from: D265A; P329A; P329G; N297A; D265A and P329A; D265A and N297A; L234 and L235A; P329A, L234A and L235A; as well as P329G, L234A and L235A. 5. An antibody according to any one of paragraphs. 1-4, where the antibody or antigen-binding fragment is characterized by negligible cell cytolysis activity. 6. An antibody according to any one of paragraphs. 1-5, wherein the antibody, or antigen-binding fragment thereof, binds with greater affinity to cells expressing higher levels of CCR7 than to cells expressing lower levels of CCR7. 7. The antibody of claim 6, wherein the antibody, or antigen-binding fragment thereof, binds with greater affinity to cancer cells that express higher levels of CCR7 than to normal cells that express lower levels of CCR7. 8. An antibody according to any one of paragraphs. 1-7, wherein the antibody, or antigen-binding fragment thereof, slightly depletes normal hematopoietic cells that express CCR7. 9. An antibody or antigen-binding fragment thereof that binds CCR7, containing: a. a heavy chain variable region that contains HCDR1 (heavy chain complementarity determining region 1) of SEQ ID NO:1, HCDR2 (heavy chain complementarity determining region 2) of SEQ ID NO:2, and HCDR3 (heavy chain complementarity determining region 3) of SEQ ID NO:3; and a light chain variable region which contains LCDR1 (light chain complementarity determining region 1) of SEQ ID NO:17, LCDR2 (light chain complementarity determining region 2) of SEQ ID NO:18, and LCDR3 (light chain complementarity determining region 3) of SEQ ID NO:19; b. a heavy chain variable region that contains HCDR1 of SEQ ID NO:4, HCDR2 of SEQ ID NO:5, and HCDR3 of SEQ ID NO:6; and a light chain variable region that contains LCDR1 of SEQ ID NO:20, LCDR2 of SEQ ID NO:21, and LCDR3 of SEQ ID NO:22; c. a heavy chain variable region that contains HCDR1 of SEQ ID NO:7, HCDR2 of SEQ ID NO:8, and HCDR3 of SEQ ID NO:9; and a light chain variable region that contains LCDR1 of SEQ ID NO:23, LCDR2 of SEQ ID NO:24, and LCDR3 of SEQ ID NO:25; d. a heavy chain variable region that contains HCDR1 of SEQ ID NO:10, HCDR2 of SEQ ID NO:11, and HCDR3 of SEQ ID NO:12; and a light chain variable region that contains LCDR1 of SEQ ID NO:26, LCDR2 of SEQ ID NO:27, and LCDR3 of SEQ ID NO:28; e. a heavy chain variable region that contains HCDR1 of SEQ ID NO:33, HCDR2 of SEQ ID NO:34, and HCDR3 of SEQ ID NO:35; and a light chain variable region that contains LCDR1 of SEQ ID NO:49, LCDR2 of SEQ ID NO:50, and LCDR3 of SEQ ID NO:51; f. a heavy chain variable region that contains HCDR1 of SEQ ID NO:36, HCDR2 of SEQ ID NO:37, and HCDR3 of SEQ ID NO:38; and a light chain variable region that contains LCDR1 of SEQ ID NO:52, LCDR2 of SEQ ID NO:53, and LCDR3 of SEQ ID NO:54; g. a heavy chain variable region that contains HCDR1 of SEQ ID NO:39, HCDR2 of SEQ ID NO:40, and HCDR3 of SEQ ID NO:41; and a light chain variable region that contains LCDR1 of SEQ ID NO:55, LCDR2 of SEQ ID NO:56, and LCDR3 of SEQ ID NO:57; h. a heavy chain variable region that contains HCDR1 of SEQ ID NO:42, HCDR2 of SEQ ID NO:43, and HCDR3 of SEQ ID NO:44; and a light chain variable region that contains LCDR1 of SEQ ID NO:58, LCDR2 of SEQ ID NO:59, and LCDR3 of SEQ ID NO:60; i. a heavy chain variable region that contains HCDR1 of SEQ ID NO:65, HCDR2 of SEQ ID NO:66, and HCDR3 of SEQ ID NO:67; and a light chain variable region that contains LCDR1 of SEQ ID NO:81, LCDR2 of SEQ ID NO:82, and LCDR3 of SEQ ID NO:83; j. a heavy chain variable region that contains HCDR1 of SEQ ID NO:68, HCDR2 of SEQ ID NO:69, and HCDR3 of SEQ ID NO:70; and a light chain variable region that contains LCDR1 of SEQ ID NO:84, LCDR2 of SEQ ID NO:85, and LCDR3 of SEQ ID NO:86; k. a heavy chain variable region that contains HCDR1 of SEQ ID NO:71, HCDR2 of SEQ ID NO:72, and HCDR3 of SEQ ID NO:73; and a light chain variable region that contains LCDR1 of SEQ ID NO:87, LCDR2 of SEQ ID NO:88, and LCDR3 of SEQ ID NO:89; l. a heavy chain variable region that contains HCDR1 of SEQ ID NO:74, HCDR2 of SEQ ID NO:75, and HCDR3 of SEQ ID NO:76; and a light chain variable region that contains LCDR1 of SEQ ID NO:90, LCDR2 of SEQ ID NO:91, and LCDR3 of SEQ ID NO:92; m. a heavy chain variable region that contains HCDR1 of SEQ ID NO:596, HCDR2 of SEQ ID NO:597, and HCDR3 of SEQ ID NO:598; and a light chain variable region that contains LCDR1 of SEQ ID NO:612, LCDR2 of SEQ ID NO:613, and LCDR3 of SEQ ID NO:614; n. a heavy chain variable region that contains HCDR1 of SEQ ID NO:599, HCDR2 of SEQ ID NO:600, and HCDR3 of SEQ ID NO:601; and a light chain variable region that contains LCDR1 of SEQ ID NO:615, LCDR2 of SEQ ID NO:616, and LCDR3 of SEQ ID NO:617; o. a heavy chain variable region that contains HCDR1 of SEQ ID NO:602, HCDR2 of SEQ ID NO:603, and HCDR3 of SEQ ID NO:604; and a light chain variable region that contains LCDR1 of SEQ ID NO:618, LCDR2 of SEQ ID NO:619, and LCDR3 of SEQ ID NO:620; or p. a heavy chain variable region that contains HCDR1 of SEQ ID NO:605, HCDR2 of SEQ ID NO:606, and HCDR3 of SEQ ID NO:607; and a light chain variable region that contains LCDR1 of SEQ ID NO:621, LCDR2 of SEQ ID NO:622, and LCDR3 of SEQ ID NO:623. 10. An antibody or antigen-binding fragment thereof that binds CCR7, according to claim 9, containing: a. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:13 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:29; b. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:45 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:61; c. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:77 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:93; or d. a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO:608; and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:624. 11. An antibody or antigen-binding fragment thereof that binds CCR7, according to claim 9, containing: a. a heavy chain containing the amino acid sequence of SEQ ID NO:15 and a light chain containing the amino acid sequence of SEQ ID NO:31; b. a heavy chain containing the amino acid sequence of SEQ ID NO:47 and a light chain containing the amino acid sequence of SEQ ID NO:63; c. a heavy chain containing the amino acid sequence of SEQ ID NO:79 and a light chain containing the amino acid sequence of SEQ ID NO:95; or d. a heavy chain containing the amino acid sequence of SEQ ID NO:610; and a light chain containing the amino acid sequence of SEQ ID NO:626. 12. An antibody according to any one of paragraphs. 1-11, where the antibody or antigen-binding fragment contains one or more cysteine substitutions. 13. An antibody according to claim 12, wherein the antibody or antigen-binding fragment thereof contains one or more cysteine substitutions selected from S152C, S375C, or both S152C and S375C, in the heavy chain of the antibody or antigen-binding fragment, where the position numbering corresponds to the EU system. 14. An antibody according to any one of paragraphs. 1-13, wherein said antibody is a monoclonal antibody. 15. An antibody-drug conjugate for use in the treatment of cancer, comprising the formula: Ab-(L-(D) _m ) _n , or a pharmaceutically acceptable salt thereof; where Ab is an antibody or an antigen-binding antigen-binding fragment thereof according to any one of paragraphs. 1-14; L is a linker; D is a drug moiety; m is an integer from 1 to 8; and n is an integer from 1 to 12. 16. The antibody-drug conjugate of claim 15, wherein said m is 1. 17. Conjugate antibodies and drugs according to any one of paragraphs. 15 or 16, where said n is from about 3 to about 4. 18. Conjugate antibodies and drugs according to any one of paragraphs. 15-17, wherein said linker is selected from the group consisting of a cleavable linker, a non-cleavable linker, a hydrophilic linker, a precharged linker, and a dicarboxylic acid linker. 19. The antibody drug conjugate of claim 18, wherein the linker is derived from a crosslinking reagent selected from the group consisting of N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl-4-(2- pyridyldithio)pentanoate (SPP), N-succinimidyl-4-(2-pyridyldithio)butanoate (SPDB), N-succinimidyl-4-(2-pyridyldithio)-2-sulfo-butanoate (sulfo-SPDB), N-succinimidyl iodoacetate ( SIA), N-succinimidyl-(4-iodoacetyl)aminobenzoate (SIAB), maleimide-PEG-NHS, N-succinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-sulfosuccinimidyl-4-(maleimidomethyl)cyclohexanecarboxylate (sulfo- SMCC) and 2,5-dioxopyrrolidin-1-yl-17-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-5,8,11,14-tetraoxo-4,7 ,10,13-tetraazaheptadecan-1-oate (CX1-1). 20. The antibody-drug conjugate of claim 18, wherein said linker has the following formula (IIA):

(IIA); where * is connected to the thiol functional group on the antibody, and ** is connected to the thiol functional group of the fragment, which is a drug; and where: L ¹ is C _1-6 alkylene, whereby one of the methylene groups may be replaced by oxygen; L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; X is -C(O)-NH-, -NHC(O)- or triazole; and alkylene is linear or branched. 21. The antibody-drug conjugate of claim 20, wherein said linker has the following formula:

where y is from 1 to 11; * is linked to a thiol functional group on the antibody, and ** is linked to a thiol functional group on the drug moiety. 22. Conjugate antibodies and drugs according to any one of paragraphs. 15-21, wherein said drug fragment is selected from the group consisting of a V-ATPase inhibitor, a proapoptotic agent, a Bcl2 inhibitor, an MCL1 inhibitor, an HSP90 inhibitor, an IAP inhibitor, an mTor inhibitor, a microtubule stabilizer, a microtubule destabilizer, auristatin, dolastatin, maytansinoid, MetAP (methionine aminopeptidase), auristatin, amanitin, pyrrolobenzodiazepine, RNA polymerase inhibitor, CRM1 nuclear export inhibitor, DPPIV inhibitor, proteasome inhibitors, inhibitors of mitochondrial phosphoryl transfer reactions, protein synthesis inhibitor, kinase inhibitor, CDK2 inhibitor, inhibitor CDK9, a kinesin inhibitor, an HDAC inhibitor, a DNA damaging agent, a DNA alkylating agent, a DNA intercalator, a DNA minor groove binding agent, and a DHFR inhibitor. 23. The antibody-drug conjugate of claim 22, wherein the cytotoxic agent is a maytansinoid. 24. The antibody-drug conjugate of claim 23 wherein the maytansinoid is N(2')-deacetyl-N(2')-(3-mercapto-l-oxopropyl)-maytansine (DM1), N(2') -deacetyl-N(2')-(4-mercapto-1-oxopentyl)-maytansine (DM3) or N(2')-deacetyl-N2-(4-mercapto-4-methyl-1-oxopentyl)-maytansine ( DM4). 25. An antibody-drug conjugate according to any one of paragraphs. 15-24 having the following formula (VIII):

where L ¹ represents C _1-6 alkylene, while one methylene group can be replaced by oxygen; L ² is C _1-6 alkylene or is -(CH ₂ CH ₂ O) _y -CH ₂ -CH ₂ - where y is from 1 to 11; X is -C(O)-NH-, -NHC(O)- or triazole; and alkylene is linear or branched; and where n is from about 3 to about 4; or a pharmaceutically acceptable salt thereof. 26. An antibody-drug conjugate for use in the treatment of cancer, having the following formula:

where n is from about 3 to about 4, and Ab is an antibody containing a heavy chain containing the amino acid sequence under SEQ ID NO:47, and a light chain containing the amino acid sequence under SEQ ID NO:63; or a pharmaceutically acceptable salt thereof. 27. Pharmaceutical composition for use in the treatment of cancer, containing an antibody-drug conjugate according to any one of paragraphs. 15-26 and a pharmaceutically acceptable carrier. 28. A method of treating or preventing cancer in a patient in need thereof, comprising administering to said patient an antibody-drug conjugate according to any one of paragraphs. 15-26 or the pharmaceutical composition of claim 27, wherein the cancer expresses CCR7. 29. The method of claim 28 wherein the antibody drug conjugate or pharmaceutical composition is administered to the patient in combination with one or more additional therapeutic compounds. 30. The method of claim 29, wherein the one or more additional therapeutic compounds are selected from a standard chemotherapeutic agent, a costimulatory molecule, or a checkpoint inhibitor. 31. The method of claim 30 wherein the costimulatory molecule is selected from OX40 agonist, CD2, CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), GITR, CD30 , CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, STING or CD83 ligand. 32. The method of claim 30 wherein the checkpoint inhibitor is selected from PD-1, PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGFR-beta inhibitor . 33. Conjugate antibodies and drugs according to any one of paragraphs. 15-26 or a pharmaceutical composition according to claim 27 for use as a drug. 34. Conjugate antibodies and drugs according to any one of paragraphs. 15-26 or a pharmaceutical composition according to claim 27 for use in the treatment or prevention of cancer expressing CCR7 in a patient in need thereof. 35. The use of an antibody or antigen-binding fragment according to any one of paragraphs. 1-14, or an antibody-drug conjugate according to any one of paragraphs. 15-26 or the pharmaceutical composition of claim 27 in the manufacture of a medicament for use in the treatment of cancer. 36. The method according to any one of paragraphs. 28-32 or the antibody-drug conjugate of claim 34, wherein the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL), peripheral T-cell lymphomas (PTCL) such as adult T-cell leukemia/lymphoma (ATLL) and anaplastic large cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL) such as mantle cell lymphoma (MCL), Burkitt's lymphoma and diffuse large B cell lymphoma (DLBCL), gastric carcinomas, non-small cell lung cancer, small cell lung cancer, head cancer and neck, nasopharyngeal carcinoma (NPC), esophageal cancer, colorectal carcinoma, pancreatic cancer, thyroid cancer, breast cancer, renal cell carcinoma, and cervical cancer. 37. The method according to any one of paragraphs. 28-32 or the antibody-drug conjugate of claim 34, wherein the cancer is selected from the group consisting of chronic lymphocytic leukemia (CLL), peripheral T-cell lymphomas (PTCL) such as adult T-cell leukemia/lymphoma (ATLL) and anaplastic large cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL) such as mantle cell lymphoma (MCL), Burkitt's lymphoma and diffuse large B cell lymphoma (DLBCL), and non-small cell lung cancer. 38. Nucleic acid that encodes an antibody or antigen-binding fragment according to any one of paragraphs. 1-14. 39. An expression vector containing a nucleic acid according to claim 38. 40. A host cell containing a vector according to claim 39 or a nucleic acid according to claim 38, while said host cell is intended to produce an antibody according to any one of paragraphs. 1-14.

Images