NZ710201B2 - Cd37-binding molecules and immunoconjugates thereof - Google Patents

Abstract

Disclosed is an antibody or antigen binding fragment thereof that specifically binds to CD37, and in vitro or ex vivo methods for depleting a B-cell comprising contacting a population of cells comprising a B-cell with said antibody or antigen binding fragment.

Description

CD37-BINDING MOLECULES AND IMMUNOCONJUGATES THEREOF Cross-Reference to Related Applications This application is a divisional application of New Zealand Application No. 615735, filed on 30 March 2012, and is related to International Patent Application No. , filed on March 2012 and claims priority from U.S. Provisional Patent Application No. 61/470,863, filed on 1 April 2011; each of which is incorporated herein by reference in its entirety.

Field of the Invention [0001a] The field of the invention generally relates to antibodies, n-binding fragments thereof, ptides, and immunoconjugates that bind to CD37, as well as to methods of using such CD37- g les for the treatment of diseases, such as autoimmune diseases and inflammatory Background of the Invention Leukocyte n CD37 ("CD37"), also known as GP52-40, tetraspanin-26, or 6, is a transmembrane protein of the tetraspanin amily (Maecker et al., 1997 FASEB J. 11:428-442).

It is a heavily glycosylated protein with four transmembrane s that is expressed on B cells during the pre-B to peripheral mature B-cell stages, but is reportedly absent on terminal differentiation to plasma cells. (Link et al., 1987, J Pathol. 152:12-21). The CD37 antigen is only weakly expressed on s, myeloid cells, and ocytes (Schwartz-Albiez et al. 1988, J. Immunol., 140(3)905-914).

However, CD37 is also expressed on malignant B-cells such as those founding non-Hodgkin’s lymphoma (NHL) and chronic lymphoid leukemia (CLL) (Moore et al. 1986, J Immunol. 137(9):3013- While the exact physiological role of CD37 is unclear, studies in CD37-deficient mice suggest an immunoregulatory function. Although mice deficient in CD37 expression have normal development (Knobeloch et al. 2000, Mol Cell Biol., :5363-9), in the C57/Bl6 background, CD37-/- T cells are hyper-proliferative (van Spriel et al., J Immunol. 172, 2953 (2004)), CD37-/- dendritic cells (DC) exhibit an increased antigen presentation (Sheng et al., Eur J Immunol. 39, 50 (2009)), and - macrophages show increased dectininduced IL-6 production (Meyer-Wentrup et al., J Immunol. 178, 154 (2007)). CD37-deficient C57/Bl6 mice also contain significantly higher level of IgA than the wild-type mice (van Spriel et al., PLoS Pathol. 5, e1000338 (2009) and Rops et al., Am J Pathol. 176, 2188 (2010)). All of these results suggest a general regulatory role of CD37 in the immune system. Interestingly, crosslinking of CD37 n by antibody on human T cells inhibits T cell proliferation induced by CD3 stimulation (van Spriel et al., J Immunol. 172, 2953 (2004)).

Antibodies are emerging as a promising method to treat human diseases including mune diseases. Currently, an anti-CD20 antibody called rituximab has been approved for - 1a - rheumatoid arthritis (RA) treatment (Edwards JC et al. 2006, Nat Rev Immunol. 6: 119). Rituximab is used in the United States in combination with methotrexate (MTX) to reduce signs and symptoms in adult patients with tely- to severely-active RA who have had an inadequate response to at least one TNF antagonist. Many studies address the use of rituximab in a variety of non-malignant autoimmune or matory disorders, including RA, in which B-cells and autoantibodies appear to play a role in disease pathophysiology. Edwards et al., Biochem Soc. Trans. 30:824-828 .

Targeting of CD20 [TEXT CONTINUES ON PAGE 2] using anti-CD20 antibody has been reported to potentially relieve signs and symptoms of a number of autoimmune or inflammatory diseases including, for example, RA ro et al., Ann. Rheum. Dis. 61 :883-888 (2002); Edwards et al., Arthritis Rheum, 46 (Suppl. 9): S46 (2002); Stahl et al., Ann. Rheum.

Dis., 62 (Suppl. 1): OP004 (2003); Emery et al., Arthritis Rheum. 48(9): S439 (2003)), lupus (Eisenberg, Arthritis. Res. Ther. 5:157-159 (2003); Leandro et al. Arthritis Rheum. 46: 2673—2677 (2002); n et al., Lupus, 13: 312—316 (2004)), immune thrombocytopenic purpura (D'Arena et al., Leuk. ma 44:561-562 (2003); Stasi et al., Blood, 98: 952-957 (2001); Saleh et al., Semin. Oncol., 27 (Supp 12):99— 103 (2000); Zaja et al., Haematologica, 87:189-195 (2002); Ratanatharathom et al., Ann. Int. Med., -279 (2000)), red cell aplasia (Auner et al., Br. J. Haematol, 116:725-728 pure (2002)); autoimmune anemia (Zaja et al., supra um appears in Haematologica 87:3 36 (2002)), cold agglutinin disease (Layios et al., Leukemia, 15:187—8 (2001); Berentsen et al., Blood, 103: 2925-2928 (2004); Berentsen et al., Br. J. Haematol., 115:79-83 (2001); Bauduer, Br. J. Haematol, 112:1083-1090 (2001); Zaja et al., Br. J. Haematol, 115:232-233 (2001)), type B syndrome of severe n resistance (Coll et al., N. Engl. J. Med., 350:310-311 (2004), mixed cryoglobulinerrnia (DeVita et al., Arthrétis Rheum. 46 Suppl. 91S206/S469 (2002)), enia gravis (Zaja et al., Neurology, 55:1062-1063 (2000); Wylam et al., J. Pediatr., 143:674—677 (2003)), r’s granulomatosis (Specks et al., Arthritis & Rheumatism 44:2836—2840 ), microscopic polyangiitis (MPA), refractory pemphigus vulgaris (Dupuy et al., Arch Dermatol, 140:91—96 (2004)), dermatomyositis (Levine, Arthritis Rheum, 46 (Suppl. 9)281299 (2002)), Sjogren's me (Somer et al., Arthritis & Rheumatism, 49:394—398 (2003)), active type-II mixed cryoglobulinemia (Zaja et al., Blood, 101:3 827-3834 ), pemphigus vulgaris (Dupay et al., Arch. Dermatol, 140:91—95 (2004)), autoimmune neuropathy (Pestronk et al., J. Neurol. urg.

Psychiatry 74:485-489 (2003)), paraneoplastic opsoclonus—myoclonus syndrome (Pranzatelli et al.

Neurology 60 (Suppl. 1) POS.128:A395 (2003)), and relapsing—remitting multiple sclerosis .

Cross et a1. (abstract) "Preliminary Results from a Phase II Trial of Rituximab in MS" Eighth Annual g of the Americas Committees for Research and Treatment in Multiple Sclerosis, 20-21 (2003).

In animal models, B—cell depletion using antibodies t B—cell antigens such as CD20 has been shown to inhibit or ameliorate several autoimmune diseases including systemic lupus ematosus (SLE), experimental autoimmune encephalomyelitis (EAE; mouse model of multiple sclerosis), type-1 diabetes (TlD) and rheumatoid arthritis (RA). Rituximab has been shown to deplete both ant and normal B cells in vivo in animal models as well as patients {Malone}; DG et al, Blood. l994;84(8):2457~66; Reff ME, et al. Blood. 3(2):435-45; Schroder C, et at. Transpl lmmunol. 2(l):l9—28). It can aiso deplete nermal B»cells from human peripheral blood rnononnclear celis (PBMCS) in in with; ments (Vugmeyster Y, et a1, Cytometry A. 2003;52(2):101—9; Vngmeyster Y and Hewell Kint lmrntmopharmacol. 2004;4(8):l l l7~24).

Campath-lH (alumtuzumab), an anti-CD52 ic IgGl, binds to the CD52 antigen, which is highly expressed on all lymphocytes (Ginaldi L, et al,Leuk Res. 1998 (2):l85-91; Hale G, et al, Tissue Antigens. 1990 Mar;35(3):l 1 8-27). It is used in patients to deplete malignant lymphocytes and is approved for treating chronic lymphocytic leukemia. It has also shown efficacy in treating multiple sclerosis and is currently in Phase III clinical g (N Engl J Med 2008; 359:1786-1801; ClinicalTrials.gov 30348 & NCT00548405). It has been shown to deplete normal lymphocytes in vitro as well (Hale G, et al. Blood. 1983 Oct;62(4):873—82; Waldmann H and Hale G Philos Trans R Soc Lond B Biol Sci. 2005 Sep 29;360(1461):1707-1l).

CD37-binding agents are also being tested as potential therapeutics for B-cell malignancies.

Emergent Biosolutions (formerly Trubion Pharmaceuticals) developed the CD37-binding agents SMIP- 016 and TRU-016 (Zhao et al., 2007, Blood, 69—2577). SMIP—016 is a single chain polypeptide that includes variable regions from a hybridoma and engineered human constant regions. TRU-016 is a humanized version of the anti-CD37 SMIP protein. See e.g. U.S. Published Application No. 2007/0059306. TRU—Ol6 is being tested ally for the treatment of c lymphocytic leukemia (CLL). Boehringer Ingelheim has also disclosed a CD37 binding agent in International Published Application No. . However, no CDC activity has been bed for any of these binding agents and no in vitro pro-apoptotic ty has been described in the e of cross-linking agents.

Radio—immunotherapy (RIT) has been attempted using a radio—labeled anti-CD37 antibody MB—l in two separate trials. Therapeutic doses of 131I-MB-l were administered to six relapsed NHL patients (Press et al. 1989 J Clin Oncol. 7(8):1027—38; Press at el. 1993, N Engl J Med. ):l2l9-24).

All six patients achieved a complete ion (CR) with a duration of four to thirty—one months. In another trial, 131I-MB—l was administered to ten relapsed NHL patients (Kaminski et al. 1992 J Clin Oncol. lO(ll):l696-7ll). A total of four patients had a response ranging in duration from two to six months, gh only one CR was reported. However, not all patients could be treated due to an unfavorable tribution of the radio—label which raised concern for radiation exposure of vital non- target organs. Indeed, RIT related toxicities were observed in these trials including severe myelosupression and cardiopulmonary toxicity. While these clinical data suggest that anti—CD37 radio- immunoconjugates may be effective, these therapies are cumbersome to administer, and at relapse post- RIT patients cannot be retreated with RIT due to the risks associated with high doses of radiation.

To me the limitations of RIT, antibody—cytotoxic agent conjugates (ACC), also called antibody-drug conjugates (ADC), have been ped. These are immunoconjugates that e a cytotoxic agent covalently linked to an antibody through a chemical linker which can allow for specific delivery of cytotoxic drugs to cells expressing a protein ized by the antibody. However, proteins that are poorly internalized are not considered to be favorable targets for such therapeutics. CD37 is structurally similar to CD20 as both antigens contain four transmembrane domains, although CD20 is not part of the tetraspanin family (Tedder et a1. 1989, J. Immun. 142: 2560—2568). Antibodies against several B-cell antigens ing CD37 and CD20 have been d for their ability to undergo endocytosis and degradation (Press et al. 1989, Cancer Res. :4906—12, and Press et a1. 1994, Blood. 83(5):1390-7).

The D37 antibody MB-l was retained on the cell surface and internalized slowly in Daudi lymphoma cells in vitro. The MB—l antibody also had a low rate of endocytosis and intracellular metabolism in NHL patient cells in vitro. Similar results were obtained with the anti—CD20 antibody 1F5, which was also retained mainly on the lymphoma cell surface and alized poorly. ADCs of CD20 antibodies have been studied previously but have not demonstrated significantly strong potency, especially when non-disulfrde or acid stable linkers are used (see for example Polson et al., 2009, Cancer Res, 69(6):2358-2364). In light of these observations, CD37 has not been considered a favorable target for antibody-drug conjugates.

While their role in cancer treatment has been studied, the potential effect of CD37-directed therapies such as antibodies, antibody tives or radio-immunoconjugates on cells involved in autoimmune diseases, inflammatory es or other disorders of the immune system is not well understood. Furthermore, none of the compounds bed above have been demonstrated to induce depletion of target cells involved in manifestation or progression of these types of diseases.

Therefore, there exists a need for CD37 binding agents including antibodies, antigen-binding fragments thereof, and antibody-drug conjugates (immunoconjugates) as a means to treat autoimmune es, inflammatory diseases, or other disorders of the immune system. The present invention addresses that need.

BRIEF SUMMARY OF THE INVENTION [0012| In one aspect, the t disclosure provides a method for depleting B-cells or treating a disease associated with aberrant B-cell activity, comprising administering to a patient an effective amount of a humanized CD37 targeting antibody or immunoconjugate provided herein. In some embodiments, the B—cells are non-cancerous FEE-cells. In some embodiments, the B—cells do not overexpress CD37.

In certain embodiments, the disease ated with aberrant B-cell activity is a disease associated with B-cell autoantibody production, and/or a disease associated with inappropriate T-cell stimulation in connection with a B—cell y.

In certain embodiments, the e characterized by autoantibody production is rheumatoid arthritis, multiple sclerosis, type I diabetes mellitus, idiopathic atory myopathy, systemic lupus erythematosus (SLE), myasthenia gravis, s disease, derrnatomyositis, polymyositis, or other autoimmune diseases, In certain embodiments, the present disclosure provides a method for depleting a B—cell comprising contacting a B-cell (e.g., in a tion of cells comprising a non-cancerous B—cell) with an antibody or antigen binding fragment thereof that specifically binds to CD37, n the antibody or fragment thereof is capable of inducing apoptosis in vitro in the absence of a cross—linking agent. In certain embodiments, the present disclosure provides a method for treating a patient having an autoimmune or inflammatory disease comprising administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to CD37, wherein the antibody or fragment thereof is capable of inducing apoptosis in vitro in the absence of a cross—linking agent. In some embodiments, the antibody or antigen—binding fragment f is also capable of inducing complement dependent cytotoxicity (CDC). In some embodiments, the antibody or antigen- g fragment thereof is also capable of inducing antibody dependent cell mediated xicity . In some embodiments, the antibody or antigen-binding fragment thereof has a long serum half- life.

In certain embodiments, the present disclosure provides a method for depleting a B-cell comprising ting a B-cell (e.g., in a population of cells comprising a non-cancerous B-cell) with an antibody or antigen binding fragment thereof that specifically binds to the same CD37 epitope as an antibody selected from the group consisting of: (a) an antibody comprising the polypeptide of SEQ ID X0255 and the polypeptide of SEQ ID NO:72; (b) an antibody comprising the polypeptide of SEQ ID NO:56 and the polypeptide of SEQ ID NO:73; (c) an antibody sing the polypeptide of SEQ ID \10257 and the polypeptide of SEQ ID NO:74; (d) an antibody comprising the polypeptide of SEQ ID \O:58 and the polypeptide of SEQ ID NO:74; (e) an antibody comprising the polypeptide of SEQ ID I\O:59 and the polypeptide of SEQ ID NO:75; (i) an antibody sing the polypeptide of SEQ ID \'O:60 and the polypeptide of SEQ ID NO:76; (g) an antibody comprising the polypeptide of SEQ ID N026] and the polypeptide of SEQ ID NO:77; (h) an dy comprising the polypeptide of SEQ ID \'O:62 and the polypeptide of SEQ ID NO:78; (i) an antibody comprising the polypeptide of SEQ ID \IO:63 and the polypeptide of SEQ ID NO:79; (j) an antibody sing the polypeptide of SEQ ID \‘Oz64 and the polypeptide of SEQ ID NO:80; (k) an antibody sing the polypeptide of SEQ ID XO:65 and the polypeptide of SEQ ID NO:81; (1) an antibody comprising the polypeptide of SEQ ID ROz66 and the polypeptide of SEQ ID NO:82; (m) an antibody comprising the polypeptide of SEQ ID V067 and the ptide of SEQ ID NO:83; (n) an antibody comprising the polypeptide of SEQ ID NO:68 and the polypeptide of SEQ ID NOz84; (0) an antibody sing the ptide of SEQ ID N069 and the polypeptide of SEQ ID NO:85; (p) an antibody sing the polypeptide of SEQ ID I\O:70 and the polypeptide of SEQ ID N0286; (q) an antibody comprising the polypeptide of SEQ ID NO:71 and the polypeptide of SEQ ID NO:87; and (r) an antibody comprising the polypeptide of SEQ ID NO:177 and the polypeptide of SEQ ID NO:l78,.

In n embodiments, the present disclosure provides a method for ng a patient having an autoimmune or inflammatory disease comprising administering to the patient a therapeutically effective amount of an dy or antigen-binding fragment thereof that specifically binds to the same CD37 epitope as an antibody selected from the group described above. In some embodiments, the antibody or antigen-binding fragment f competitively inhibits an antibody selected from the group described above.

In certain embodiments, the present disclosure es a method for depleting a B-cell comprising contacting a B—cell (e.g., in a population of cells comprising a non-cancerous ) with an dy or antigen-binding fragment thereof that specifically binds to CD37 and specifically binds to the polypeptide of SEQ ID NO: 184. In certain embodiments, the present disclosure provides a method for treating a patient having an autoimmune or atory disease comprising administering to the patient a therapeutically ive amount of an antibody or n—binding fragment thereof that specifically binds to CD37 and specifically binds to the polypeptide of SEQ ID NO: 184. In some embodiments, the antibody or antigen-binding fragment thereof does not bind to the polypeptide of SEQ ID NO: 185.

In certain ments, the present disclosure provides a method for depleting a B-cell comprising contacting a B—cell (e.g., in a population of cells sing a non—cancerous B-cell) with an antibody or antigen-binding fragment thereof that specifically binds to CD37 and does not specifically bind to the polypeptide of SEQ ID NO: 185. In certain embodiments, the present disclosure provides a method for treating a patient having an autoimmune or inflammatory disease comprising administering to the t a therapeutically effective amount of an antibody or n-binding fragment thereof that specifically binds to CD37 and does not specifically bind to the polypeptide of SEQ ID NO: 185.

In certain embodiments, the present disclosure provides a method for depleting a B-cell sing contacting a B-cell (e.g., in a population of cells comprising a non-cancerous ) with an antibody or antigen-binding fragment thereof produced by a hybridoma selected from the group consisting of ATCC Deposit Designation PTA-10664, ted with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10665, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10666, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10667, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation 668, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10669, deposited with the ATCC on ry 18, 2010, and ATCC Deposit Designation PTA—10670, deposited with the ATCC on February 18, 2010. In certain embodiments, the present disclosure provides a method for treating a patient having an mune or inflammatory disease comprising administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof produced by a hybridoma described above...

In certain embodiments, the present disclosure provides a method for ing a B-cell comprising contacting a B-cell (e.g., in a population of cells comprising a non—cancerous B-cell) with an antibody or antigen—binding fragment thereof that cally binds to CD37, wherein the antibody comprises polypeptide sequences selected from the group consisting of: (a) SEQ ID NOs: 4, 5, and 6 and SEQ ID NOS: 28, 29, and 30; (b) SEQ ID NOS: 7, 8, and 9 and SEQ ID NOS: 31, 32, and 33; (c) SEQ ID NOS: 10, 11, and 12 and SEQ ID NOS: 34, 35, and 36; (d) SEQ ID NOS: 13, 14, and 15 and SEQ ID NOS: 37, 38, and 39; (e) SEQ ID NOS: 13, 14, and 15 and SEQ ID NOS: 37, 40, and 39; (f) SEQ ID NOS: 16, 17, and 18 and SEQ ID NOS: 41, 42, and 43; (g) SEQ ID N05: 19, 20, and 21 and SEQ ID NOS: 44, 45, and 46; (h) SEQ ID NOS: 19, 20, and 21 and SEQ ID NOS: 44, 47, and 46; (i) SEQ ID NOS; 22, 23, and 24 and SEQ ID NOS: 48, 49, and 50; (j) SEQ ID NOS: 22, 23, and 24 and SEQ ID NOs: 48, 51, and 50; (k) SEQ ID NOS: 25, 26, and 27 and SEQ ID NOS: 52, 53, and 54; (l) SEQ ID N05: 171, 172 or 181, and 173 and SEQ ID NOS: 174, 175, and 176; (m) ts of (a) to (1) comprising 1, 2, 3, or 4 conservative amino acid tutions. In certain embodiments, the present disclosure provides a method for treating a patient having an autoimmune or inflammatory e comprising administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof with an antibody or antigen-binding fragment thereof that Specifically binds to CD37, wherein the antibody comprises polypeptide sequences selected from the group described above. In some embodiments, the antibody or antigen-binding fragment thereof comprises polypeptide sequences that are at least 90% identical to polypeptide sequences described above. In some embodiments, the polypeptide ces are at least 95% identical to the polypeptide ces. In some embodiments, the polypeptide ces are at least 99% identical to the polypeptide sequences. In some ments, the antibody or antigen-binding fragment thereof comprises polypeptide sequences that are at least 90% identical, at least 95% identical, at least 99% identical, or idential to the polypeptide sequences of SEQ ID NO: 57 and SEQ ID NO:74. In some embodiments, the antibody or n—binding fragment thereof comprises polypeptide ces that are at least 90% identical, at least 95% identical, at least 99% cal, or idential to the polypeptide sequences of SEQ ID NO: 58 and SEQ ID NO:74. In some embodiments, the antibody or antigen-binding fragment thereof comprises polypeptide sequences that are at least 90% identical, at least 95% identical, at least 99% identical, or al to the polypeptide sequences of SEQ ID NO: 63 and SEQ ID NO:79. In some ments, the antibody or antigen-binding fragment thereof comprises polypeptide sequences that are at least 90% identical, at least 95% identical, at least 99% identical, or idential to the polypeptide sequences of SEQ ID NO: 65 and SEQ ID NO:81.

In some embodiments, the antibody or antigen binding fragment thereof is murine, non- human, humanized, chimeric, resurfaced, or human.

In some embodiments, the antibody or dy fragment is e of inducing apoptosis of a cell expressing CD37 in vitro in the absence of cross-linking agents. In some embodiments, the antibody or antigen binding fragment is capable of inducing complement dependent cytotoxicity (CDC). In some embodiments, the antibody is e of inducing antibody dependent cell mediated cytotoxicity (ADCC).

In certain embodiments, the present disclosure provides a method for depleting a B-cell comprising contacting a B-cell (e.g., in a tion of cells comprising a non-cancerous B-cell) with a human or humanized antibody or antigen binding fragment thereof that specifically binds to CD37, wherein the antibody or fragment thereof is capable of ng apoptosis of a cell expressing CD37 in vitro in the absence of cross—linking agents. In certain embodiments, the present disclosure provides a method for treating a patient having an autoimmune or inflammatory e comprising administering to the patient a therapeutically effective amount of a human or humanized antibody or antigen g fragment thereof that specifically binds to CD37, wherein the dy or fragment thereof is capable of inducing sis of a cell expressing CD37 in vitro in the absence of cross-linking agents. In some embodiments, the human or humanized antibody or antigen binding fragment thereof is also capable of inducing complement dependent cytotoxicity (CDC). In some embodiments, the human or humanized antibody or antigen binding fragment thereof is also capable of inducing antibody dependent cell mediated cytotoxicity (ADCC).

In some embodiments, the antibody or antigen-binding fragment binds to human CD37 and macaque CD37.

In some embodiments, the antibody is a full length antibody. In some embodiments, an antigen-binding fragment is used. In some embodiments, the antibody or antigen—binding fragment thereof comprises a Fab, Fab', F(ab')2, F(1, single chain Fv or scFv, disulfide linked Fv, V-NAR domain, IgNar, intrabody, IgGACHZ, minibody, F(ab')3, tetrabody, triabody, diabody, single-domain antibody, DVD—lg, Fcab, mAb2, (scFv)2, or scFv-Fc.

In some embodiments, the antibody or antigen-binding nt thereof is linked via a linker (L) to a cytotoxic agent (C) to form an immunoconjugate.

In certain embodiments, the present disclosure provides a method for depleting a B—cell comprising contacting a B-cell (e.g., in a population of cells comprising a ncerous B-cell) with a ition comprising an immunoconjugate having the a (A) - (L) - (C), wherein: (A) is an antibody or antigen binding fragment that specifically binds to CD37; (L) is a eavable linker; and (C) is a cytotoxic agent; and wherein the linker (L) links (A) to (C). In certain embodiments, the present disclosure provides a method for treating a patient having an autoimmune or inflammatory disease comprising administering to the patient a therapeutically effective amount of a composition comprising an conjugate having the formula (A) — (L) - (C), wherein: (A) is an dy or antigen binding fragment that ically binds to CD37; (L) is a non-cleavable linker; and (C) is a cytotoxic agent; and n the linker (L) links (A) to (C). In some embodiments, the immunoconjugate has a serum half—life that is comparable to that of the naked antibody, In certain embodiments, the present disclosure provides a method for depleting a B-cell comprising contacting a B-cell (e.g., in a population of cells comprising a non-cancerous B-cell) with a composition comprising an immunoconjugate having the formula (A) - (L) — (C), wherein: (A) is an antibody or antigen binding fragment that specifically binds to CD37; (L) is a linker; and (C) is a maytansinoid; and wherein the linker (L) links (A) to (C). In n embodiments, the present disclosure provides a method for treating a patient having an autoimmune or inflammatory disease comprising administering to the patient a therapeutically effective amount of a composition comprising an immunoconjugate having the formula (A) - (L) - (C), wherein: (A) is an antibody or antigen binding fragment that specifically binds to CD37; (L) is a linker; and (C) is a maytansinoid; and wherein the linker (L) links (A) to (C).

In some embodiments, the linker is a non-cleavable linker. In some embodiments, the conjugate further comprises a second (C). In some ments, the immunoconjugate further comprises a third (C). In some embodiments, the conjugate further comprises a fourth (C). In some embodiments, the immunoconjugate comprises 2—6 (C). In some embodiments, the immunoconjugate comprises 3-4 (C).

In some ments, the linker is selected from the group consisting of a ble linker, a non—cleavable linker, a hydrophilic linker, and a dicarboxylic acid based linker. In some embodiments, the linker is selected from the group ting of: N—succinimidyl 4-(2—pyridyldithio)pentanoate (SPP); N—succinimidyl 4-(2—pyridyldithio)butanoate (SPDB) or N-succinimidyl 4-(2-pyridyldithio)—2- sulfobutanoate (sulfo-SPDB); N-succinimidyl 4—(maleirnidomethyl) cyclohexanecarboxylate (SMCC); N- sulfosuccinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (sulfoSMCC); N—succinimidyl (iodoacetyl)—aminobenzoate (SIAB); and N—succinimidyl—[(N-maleimidopropionamido)— tetraethyleneglycol] ester (NHS-PEG4-maleimide). In some ments, the linker is inimidyl— [(N-maleimidopropionamido)—tetraethyleneglycol] ester (NHS-PEG4-maleimide).

In some embodiments, the cytotoxic agent is ed from the group consisting of a maytansinoid, maytansinoid analog, doxornbicin, a modified doxorubicin, benzodiazepine, taxoid, CC— 1065, CC-lO65 analog, duocarznycin, duocarznycin analog, calicheamicin, dolastatin, dolastatin analog, aristatin, tomaymycin derivative, and leptomycin derivative or a g of the agent. In some embodiments, the cytotoxic agent is a sinoid. In some embodiments, the cytotoxic agent is N(2')— deacetyl-N(2')-(3-mercapto—l-oxopropyl)—maytansine (DMl) or N(2')—deacetyl-N2—(4-mercapto—4-methyl— entyl)—maytansine (DM4).

In some embodiments, the composition sing an immunoconjugate ses multiple cytotoxic agents (C) with an average of about 3 to about 4 (C) per (A). In some embodiments, the irrirnunoeonjugates have an average of about 3.5 (C) per (A). In some embodiments, the immunoccnjugates have an e ofabout 3.5 i 0.5 (C) per (A).

. In some embodiments, the composition comprising an immunoconjugate comprises an antibody comprising SEQ ID NO:57 and SEQ ID NO:74 or SEQ ID NO:58 and SEQ ID NO:74, an SMCC linker, and DM1. In some embodiments, the composition comprising an conjugate comprises an antibody comprising SEQ ID NO:63 and SEQ ID NO:79, an SMCC linker, and DM1. In some embodiments, the composition comprising an conjugate comprises an antibody comprising SEQ ID NO:65 and SEQ ID NO:81, an SMCC linker, and DM1.

In some embodiments, the antibody or n-binding fragment is capable of depleting B- cells. In some embodiments, the antibody or antigen—binding fragment is capable of inhibiting T-cell responses.

In some embodiments, the B-cell is in a composition further comprising a T-cell. In some ments, the B—cell is in a ition comprising peripheral blood mononuclear cells. In some embodiments, the peripheral blood mononuclear cells were obtained from a human. In some embodiments, the B—cell is in whole blood. In some embodiments, the whole blood was obtained from a human. In some embodiments, the B—cell is in an organism. In some embodiments, the B-cell is in a t having an autoimmune or inflammatory disease.

In some embodiments, the B-cell is an autoreactive B—cell.

In some embodiments, at least about 30% of B-cells are depleted. In some embodiments, less than about 5% of T-cells are ed.

In some embodiments, a second therapeutic agent is administered. In some embodiments, the second eutic is selected from the group consisting of methotrexate, an anti—CD20 therapeutic, an L-6 receptor therapeutic, ar anti-IL—12/23p40 therapeutic, a chemotherapeutic, an immunosuppressant, an nterferon beta-1a therapeutic, glatiramer acetate, an anti-0L4—integrin eutic, fingolimod, an anti-BLys therapeutic, CTLA-Fc, or an anti-TNF therapeutic. In some ments, the second therapeutic is an antibody directed against an antigen selected from a group consisting of CD3, CD14, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD36, CD3 8, CD40, CD44, CD52, CD55, CD59, CD56, CD70, CD79, CD80, CD103, CD134, CD137, CD138, and CD152. In some embodiments, the second therapeutic is an dy directed against an antigen selected from the group consisting of IL-2, IL—6, IL—12, IL-23, IL—12/23 p40, IL—17, IFNy, TNFoc, IFNoc, IL—15, IL-21, IL—Ia, IL- lb, IL—18, IL-8, IL—4, GM—CSF, IL—3, and IL—5.

In some embodiments, the autoimmune or inflammatory disease is selected from the group consisting of rheumatoid arthritis, multiple sclerosis, type I diabetes mellitas, idiopathic inflammatory myopathy, systemic lupus erythematosus (SLE), myasthenia gravis, Grave's disease, dermatomyositis, polymyositis, s diasease, ulcerative colitis, gastritis, Hashimoto’s thyroiditis, asthma, psoriasis, psoriatic arthritis, dertmatitis, systemic dema and sclerosis, inflammatory bowel e (IBD), respiratory distress syndrome, meningitis, encephalitis, uveitis, glmerulonephritis, eczema, atherosclerosis, yte adhesion deficiency, Raynaud’s syndrome, Sjögren’s syndrome, Reiter’s disease, Beheet’s disease, immune complex nephritis, IgA nephropathy, IgM polyneuropathies, immune-mediated thrombocytopenias, acute idiopathic thrombocytopenic a, chronic idiopathic thrombocytopenic purpura, hemolytic anemia, myasthenia gravis, lupus nephritis, atopic dermatitis, pemphigus is, opsoclonus-myoclonus me, pure red cell aplasia, mixed cryoglobulinemia, ankylosing spondylitis, hepatitis C-associated cryoglobulinemic vasculitis, chronic focal encephalitis, bullous pemphigoid, ilia A, membranoproliferative ulonephritis, adult and juvenile dermatomyositis, adult polymyositis, chronic urticaria, primary biliary cirrhosis, neuromyelitis optica, ’ dysthyroid disease, bullous pemphigoid, membranoproliferative glomerulonephritis, Churg- Strauss syndrome, juvenile onset es, hemolytic anemia, atopic dermatitis, systemic sclerosis, Sjögren’s syndrome and glomerulonephritis, omyositis, eutrophil cytoplasmic dy (ANCA), aplastic anemia, autoimmune hemolytic anemia (AIHA), factor VIII deficiency, hemophilia A, autoimmune neutropenia, Castleman's syndrome, Goodpasture's syndrome, solid organ transplant rejection, graft versus host disease (GVHD), autoimmune hepatitis, lymphoid interstitial pneumonitis, HIV, bronchiolitis obliterans (non-transplant), Guillain-Barre Syndrome, large vessel vasculitis, giant cell asu's) arteritis, medium vessel vasculitis, Kawasaki's Disease, polyarteritis nodosa, Wegener’s granulomatosis, microscopic polyangiitis (MPA), Omenn’s syndrome, chronic renal failure, acute infectious mononucleosis, HIV and herpes virus associated diseases. [0039a] tions of the specific embodiments of the invention as claimed herein follow. [0039b] According to a first embodiment of the invention, there is ed an isolated dy or antigen binding nt thereof that specifically binds to CD37, wherein said antibody or fragment thereof comprises the heavy chain and light chain variable region CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 171, 172 or 181, and 173 and SEQ ID NOs: 174, 175, and 176, respectively. [0039c] According to a second embodiment of the invention, there is provided an isolated cell producing the antibody or n binding fragment thereof of the first embodiment. [0039d] According to a third embodiment of the invention, there is provided a method of making an anti-CD37 antibody or antigen-binding fragment f comprising (a) culturing the cell of the second embodiment; and (b) isolating said antibody, or antigen binding fragment thereof from said cultured cell. [0039e] According to a fourth embodiment of the invention, there is provided an immunoconjugate having the formula (A) - (L) - (C), wherein: (A) is the antibody or antigen g fragment thereof as defined in the first embodiment; (L) is a linker; and (C) is a cytotoxic agent; and n said linker (L) links (A) to (C). [0039f] According to a fifth embodiment of the invention, there is ed an in vitro or ex vivo method for depleting a B-cell comprising contacting a population of cells comprising a B-cell with the - 11a - dy or antigen binding fragment of the first embodiment or the immunoconjugate of the fourth ment.

BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES Figure 1 depicts an FL2-H (PE) histogram overlay for a flow cytometry experiment with human B-cells. The following conditions are shown: antibody control (dark ), isotype control stain (light filled), anti-CD37 stain (thick black line), and anti-CD20 stain (dashed line) for CD19+ B-cells.

Figure 2 depicts the results of in vitro depletion experiments using purified human PBMC samples d with 10 μg/mL of huCD37-3, huCD37SMCC-DM1, huCD37-50, huCD37 SMCC-DM1, rituximab, TRU-016, or alemtuzumab. Results from two different donors are shown in panel A and B.

Figure 3 depicts the s of in vitro depletion experiments using purified human PBMC samples d with varying concentrations of huCD37SMCC-DM1. Results from two different donors are shown in panels A and B. Figure 3 (C) shows the results using huCD37-3, huCD37-38, huCD37-50 and huCD37-56.

Figure 4 depicts the results of in vitro depletion experiments using unpurified whole human blood samples treated with 10 μg/mL of huCD37-3, SMCC-DM1, huCD37-50, huCD37 SMCC-DM1, rituximab, TRU-016, or alemtuzumab.

[TEXT CONTINUES ON PAGE 12] {new Figure 5 depicts the results of in vine depletion experiments using fied whole human blood samples d with vaming concentrations of (A) hnCDB 7-3, huClIXW—B—Sl‘vfiiCC~li)lVll, and inab and (B) huCDBY-IB, huCDW—lSMCC-DMI , huCD37«50, and ritnximab, EQMSE Figure 6 depicts release of lFN--y (Interferon), "l‘NF~€x (Tumor is Factor) and Ibo (lnterleukimé) measured by ELlSpot as number of spots per leGEZS peripheral blood mononuclear cells (PBMCS) from one healthy human donor incubated for ill—20 hours with compounds at a concentration of 2,5 rig/ml;- to 25$ gig/nil... {0046} Figure ‘7 depicts release of 119wa (Interferon), TNF~«:L {Tumor Necrosis Factor) and lire (interleukineé) measured by ELlSpot as mimber of spots per 5x"? peripheral blood mononuclear cells (PBMCs) from a second healthy human donor incubated for 18-20 hours with compounds at a concentration of 2.5 ng/mL to 250' ug/mL.

Figure 8 depicts the binding curve of anti-muCD37 monoclonal antibody clone 252—3.

Figure 9 shows the activity of the 252—3 antibody in depleting peripheral blood B cells (A) and in inhibiting EAE (B) in 6 mice. In (A), each symbol represent one mouse; to compare the B cell level in l vs. experimental mice, B cell level was normalized with T cell level and ratio of B/T cell in control mice was considered 100%. In (B), open and closed s represent mean of EAE score in control group (n=10) and 252—3 antibody treated group (n=10), respectively; arrow indicates day of antibody injection.

Figure 10 shows the activity of the 252-3 antibody in depleting peripheral blood B cells (A) and in ting TlD (B) in NOD mice. In (A), each symbol represent one mouse; to compare the B cell level in control vs. experimental mice, B cell level was normalized with T cell level and ratio of B/T cell in control mice was considered 100%. In (B), open and closed symbols ent the diabetes incidence in control group (n36) and 252—3 antibody treated group (n=6), respectively.

Figure 11 shows the activity of the 252-3 dy in depleting eral blood B cells (A) and in inhibiting CIA (B) in DBA/l mice. In (A), each symbol represent one mouse; to compare the B cell level in control vs. experimental mice, B cell level was ized with T cell level and ratio of B/T cell in control mice was considered 100%. In (B), open and closed symbols represents mean of CIA score in control group (n=l2) and 252-3 antibody treated group (n:12), respectively; arrow indicates day of antibody injection.

DETAILED DESCRIPTION OF THE INVENTION {GoSE} The present invention provides methods of depleting B~cells and of treating diseases associated with aberrant Bucell activity and/or aberrant 'l‘~cell stimulation in connection with a B~cell pathway using (2937 binding molecules.

I. Definitions To facilitate an understanding of the present ion, a number of terms and phrases are defined below.

The term CD37 as used herein, refers to any native CD37, unless otherwise indicated. CD37 is also referred to as GP52-40, leukocyte antigen CD37, and Tetraspanin—26. The term "CD37" encompasses "full-length," unprocessed CD37 as well as any form of CD37 that results from processing in the cell. The term also encompasses naturally occurring variants of CD37, e.g., splice variants, allelic variants, and isoforms. The CD37 polypeptides described herein can be isolated from a variety of sources, such as from human tissue types or from another , or ed by recombinant or synthetic methods.

The term "antibody" means an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or combinations of the foregoing through at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term "antibody" encompasses intact polyclonal antibodies, intact monoclonal antibodies, antibody fragments (such as Fab, Fab’, F(ab')2, and Fv fragments), single chain Fv (scFv) mutants, multispecific dies such as bispecific antibodies ted frorre at least two intact antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising an antigen determination portion of an antibody, and any other modified immunoglobulin molecule comprising an antigen recognition site so long as the antibodies exhibit the desired biological activity. An dy can be of any the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or subclasses (isotypes) thereof (e.g. lgGl, lgG2, IgGE, IgG4, lgAl and IgAZ), based on the identity of their heavy-chain nt domains referred to as alpha, delta, epsilon, gamma, and mu, respectively. The ent s of immunoglobulins have different and well known subunit structures and three-dimensional configurations. Antibodies can be naked or conjugated to other molecules such as toxins, sotopes, etc.

A "blocking" antibody or an "antagonist" antibody is one which inhibits or reduces ical activity of the antigen it binds, such as CD37. in some embodiments, ng antibodies or antagonist antibodies substantially or completely inhibit the biological activity of the antigen. The biological activity can be reduced by 10%, 20%, 30%, 50%, 70%, 80%, 90%, 95%, or even 100%.

The term "anti-CD37 antibody" or "an antibody that binds to CD3 7" refers to an antibody that is capable of binding CD37 with sufficient y such that the antibody is useful as a diagnostic and/or therapeutic agent in targeting CD37. The extent of g of an D37 dy to an unrelated, non- CD37 protein can be less than about 10% of the g of the antibody to CD37 as measured, e.g., by a radioimmunoassay (RIA). In certain embodiments, an antibody that binds to CD37 has a dissociation constant (Kd) of :1 nM, £100 nM, 510 nM, 51 nM, or 30.1 nM.

The term ody fragment" refers to a portion of an intact antibody and refers to the antigenic determining le regions of an intact antibody. Examples of antibody fragments include, but are not limited to Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, single chain antibodies, and pecific antibodies formed from antibody nts.

A "monoclonal antibody" refers to a homogeneous antibody population involved in the highly specific recognition and binding of a single nic determinant, or epitope. This is in contrast to polyclonal antibodies that typically e different antibodies directed against different antigenic determinants. The term "monoclonal antibody" encompasses both intact and full—length monoclonal antibodies as well as antibody nts (such as Fab, Fab', F(ab')2, Fv), single chain (scFv) mutants, fusion proteins sing an antibody portion, and any other modified immunoglobulin molecule comprising an antigen recognition site. Furthermore, "monoclonal antibody" refers to such antibodies made in any number of manners including but not limited to by oma, phage selection, inant expression, and enic animals.

The term ized antibody" refers to forms of non—human (e.g. murine) antibodies that are specific immunoglobulin chains, chimeric immunoglobulins, or fragments thereof that contain minimal non-human (e.g., murine) sequences. Typically, humanized antibodies are human immunoglobulins in which residues from the complementary determining region (CDR) are ed by residues from the CDR of a non-human species (e.g. mouse, rat, rabbit, hamster) that have the desired specificity, affinity, and capability (Jones et al., 1986, Nature, 321:522-525; Riechmann et al., 1988, Nature, 332:323-327; Verhoeyen et al., 1988, Science, 239:1534-1536). In some instances, the Fv framework region (FR) residues of a human immunoglobulin are replaced with the corresponding residues in an dy from a man species that has the desired specificity, affinity, and capability. The humanized antibody can be r modified by the substitution of additional residues either in the Fv framework region and/or within the replaced non-human residues to refine and optimize antibody specificity, affinity, and/or capability. In general, the humanized antibody will comprise substantially all of at least one, and typically two or three, variable domains containing all or ntially all of the CDR regions that correspond to the non-human immunoglobulin whereas all or substantially all of the FR s are those of a human immunoglobulin consensus sequence. The humanized antibody can also comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. Examples of methods used to generate humanized antibodies are described in US. Pat. 5,225,539. {0060} A "variabie region" oft-1n antibody refers tn the variable regien of the dy Eight. chain or the variable region of the antibody heavy chain, either aicne er in combination. The variable s of the heavy and Eight chain each consist at" four framework regions (FR) cnnnected by three complementarity determining regions {CDRs} also knnwn as hypervariabie regicns. The CDRs in each chain are held together in close proximity by the FRs and, with the CDRs from the other chain, contrébute to the formation of the antigen—binding site of antibodies. There are at least two ques for determining CDRs: (1) an approach based on cross—species sequence variability (i.e., Kabat et al.

Sequences of Proteins of Immunological Interest, (5th ed., 1991, National Institutes of Health, Bethesda Md.)); and (2) an approach based on crystallographic studies of n-antibody complexes (Al-lazikani et a1 (1997) J. Molec. Biol. 273:927-948)). In addition, combinations of these two approaches are sometimes used in the art to determine CDRs.

The Kabat numbering system is generally used when referring to a residue in the variable domain (approximately es 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g, Kabat et al., Sequences of logical Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

The amino acid position ing as in Kabat, refers to the numbering system used for heavy chain variable domains or light chain variable domains of the compilation of dies in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Using this numbering system, the actual linear amino acid sequence can contain fewer or additional amino acids ponding to a shortening of, or insertion into, a FR or CDR of the variable domain. For example, a heavy chain variable domain can include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g. residues 82a, 82b, and 82c, etc according to Kabat) after heavy chain FR residue 82. The Kabat ing of residues can be determined for a given antibody by alignment at regions of gy of the sequence of the antibody with a "standard" Kabat numbered sequence. Chothia refers instead to the location of the structural loops (Chothia and Lesk J. Mol. Biol. 1962901-917 (1987)). The end of the Chothia CDR—HI loop when numbered using the Kabat ing convention varies between H32 and H34 depending on the length of the loop (this is because the Kabat ing scheme places the insertions at H35A and H3 5B; if neither 35A nor 35B is t, the loop ends at 32; if only 35A is present, the loop ends at 33; if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable s represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular's AbM antibody modeling software.

Loop Kabat AbM Chothia LE L24FL34 L24—L34 L24—L34 LE. 1450-1456 LEO-L56 L50—L56 L3 L89—L97 L89—L97 L89—L97 H1 BBL-H358 H26—H3SB H26—H32..34 (Kabat Numbering) H1 1131-1135 1126—1135 H26-H32 (Chothia Numbering) H2 ESQ-H65 158 HS2-1156 H3 11931-1102 11954-1132 1195—11102 The term "human antibody" means an antibody produced by a human or an antibody having an amino acid sequence corresponding to an antibody produced by a human made using any technique known in the art. This definition of a human antibody includes intact or full—length antibodies, fragments thereof, and/or antibodies comprising at least one human heavy and/or light chain ptide such as, for example, an antibody comprising murine light chain and human heavy chain ptides.

The term ric antibodies" refers to antibodies wherein the amino acid sequence of the immunoglobulin le is derived from two or more species. Typically, the variable region of both light and heavy chains corresponds to the variable region of antibodies derived from one species of mammals (e.g. mouse, rat, , etc) with the desired specificity, affinity, and capability while the constant regions are homologous to the sequences in antibodies derived from another (usually human) to avoid ing an immune response in that species.

The term "epitope" or "antigenic determinant" are used interchangeably herein and refer to that portion of an antigen capable of being recognized and specifically bound by a particular antibody.

When the antigen is a polypeptide, es can be formed both from uous amino acids and noncontiguous amino acids juxtaposed by tertiary folding of a protein. Epitopes foréned from uous amino acids are lly retained upon protein denaturing, whereas epitopes formed by tertiary folding are typically lost upon protein ring. An epitope typically includes at least 3, and more y, at least 5 or 8-10 amino acids in a unique l conformation.

"Binding affinity" generally refers to the th of the sum total of noncovalent interactions between a single binding site of a le (e.g., an antibody) arzd its binding partner (e.g., an antigen).

Unless indicated ise, as used herein, "binding affinity" refers to intrinsic binding affinity which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high—affinity antibodies -17_ generally bind antigen faster and tend to remain bound longer. A variety of methods of measuring g affinity are known in the art, any of which can be used for purposes of the present invention. Specific illustrative embodiments are described in the following.

"Or better" when used herein to refer to binding affinity refers to a stronger binding between a molecule and its binding partner. "Or better" when used herein refers to a er binding, represented by a smaller numerical Kd value. For example, an antibody which has an affinity for an antigen of "0.6 nM or better", the antibody's y for the antigen is <0.6 nM, i.e. 0.59 nM, 0.58 nM, 0.57 nM etc. or any value less than 0.6 HM.

By "specifically binds," it is lly meant that an dy binds to an epitope via its antigen binding domain, and that the bindirgg entails some complementarity between the antigen binding domain and the e. According to this definition, an antibody is said to "specifically bind" to an epitope when it binds to that epitope, via its antigen binding domain more readily than it would bind to a random, unrelated epitope. The term "specificity" is used herein to qualify the relative affinity by which a n antibody binds to a certain e. For example, antibody "A" may be deemed to have a higher specificity for a given e than antibody "B," or antibody "A" may be said to bind to epitope "C" with a higher city than it has for related epitope "D." By "preferentially binds," it is meant that the antibody specifically binds to an e more readily than it would bind to a related, similar, homologous, or analogous epitope. Thus, an antibody which "preferentially binds" to a given epitope would more likely bind to that epitope than to a related epitope, even though such an antibody may cross-react with the related epitope.

An antibody is said to "competitively inhibit" binding of a reference antibody to a given epitope if it preferentially binds to that e to the extent that it blocks, to some degree, binding of the reference antibody to the epitope. Competitive inhibition may be determined by any method known in the art, for example, competition ELISA . An antibody may be said to itively inhibit binding of the nce antibody to a given epitope by at least 90%, at least 80%, at least 70%, at least 60%, or at least 50%.

The phrase "substantially similar," or "substantially the same", as used herein, denotes a sufficiently high degree of similarity between two numeric values (generally one associated with an antibody of the invention and the other associated with a reference/comparator antibody) such that one of skill in the art would consider the difference n the two values to be of little or no biological and/or statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values). The difference between said two values can be less than about 50%, less than about 40%, less than about 30%, less than about 20%, or less than about 10% as a function of the value for the reference/comparator antibody.

A polypeptide, dy, polynucleotide, vector, cell, or composition which is "isolated" is a polypeptide, antibody, polynucleotide, vector, cell, or composition which is in a form not found in nature. ed polypeptides, antibodies, polynucleotides, vectors, cell or compositions include those which have been purified to a degree that they are no longer in a form in which they are found in nature. In some embodiments, an dy, cleotide, vector, cell, or composition which is ed is substantially pure.

As used herein, "substantially pure" refers to material which is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.

The term "immunoconjugate" or "conjugate" as used herein refers to a compound or a tive thereof that is linked to a cell binding agent (i.e., an anti—CD37 antibody or fragment thereof) and is defined by a generic formula: C—L-A, wherein C — cytotoxin, L = , and A = cell binding agent or anti-CD37 antibody or antibody fragment. Immunoconjugates can also be defined by the generic formula in reverse order: A-L-C.

A "linker" is any chemical moiety that is capable of linking a compound, usually a drug, such thereof in a stable, as a maytansinoid, to a cell-binding agent such as an anti CD37 antibody or a fragment covalent manner. Linkers can be susceptible to or be substantially resistant to acid—induced cleavage, light—induced cleavage, peptidase-induced ge, esterase-induced cleavage, and disulfide bond cleavage, at conditions under which the compound or the antibody remains . Suitable linkers are well known in the art and include, for example, disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups. Linkers also include charged linkers, and hydrophilic forms thereof as bed herein and know in the art.

The terms "cancer" and "cancerous" refer to or describe the physiological ion in mammals in which a tion of cells are characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, ma, sarcoma, and leukemia. " and "neoplasm" refer to one or more cells that result from excessive cell growth or proliferation, either benign (noncancerous) or malignant (cancerous) including pre-cancerous lesions. Examples of "cancer" or "tumorigenic" diseases which can be treated and/or prevented include B-cell lymphomas including NHL, B-cell neoplasms, such as B-cell chronic precursor B—cell lymphoblastic leukemia/lymphoma and mature lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL), B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, mantle cell lymphoma (MCL), follicular lymphoma (FL), including low— grade, intermediate—grade and rade FL, cutaneous follicle center lymphoma, marginal zone B-cell lymphoma (MALT type, nodal and splenic type), hairy cell leukemia, diffuse large B-cell lymphoma, Burkitt's lymphoma, plasmacytoma, plasma cell myeloma, post-transplant proliferative disorder, and anaplastic large—cell lymphoma (ALCL). Non-cancerous cells are cells that do not result in the "l9— fonnation of tumors or neoplasms or the pment of cancer. However, non—cancerous cells can contribute to disease, e.g., autoimmune dsieases, and include, for example auto-reactive B-cells.

The terms "cancer cell," "tumor cell," and grammatical equivalents refer to the total tion of cells derived from a tumor or a ncerous lesion, including both non-tumorigenic cells, which comprise the bulk of the tumor cell population, and tumorigenic stem cells (cancer stem cells). As used herein, the term "tumor cell" will be modified by the term "non-tumorigenic" when referring solely to those tumor cells lacking the capacity to renew and differentiate to distinguish those tumor cells from cancer stem cells.

The term "autoreactive" refers to a cell, tissue, protein, dy or other substance that produces an immune response directed against an organism's own cells, tissues, proteins, antibodies, or other substances.

The term "subject" refers to any animal (e.g., a mammal), including, but not limited to humans, non-human primates, rodents, and the like, which is to be the recipient of a particular treatment.

Typically, the terms "subject" and n " are used interchangeably herein in nce to a human subject.

Administration "in combination with" one or more farther therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order.

The term "pharmaceutical formulation" refers to a preparation which is in such form as to permit the ical activity of the active ingredient to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the thrrnulation would be administered.

The formulation can be sterile. {0082] An "effective amount" of an antibody as disclosed herein is an amount sufficient to carry out a specifically stated e. An "effective amount" can be determined empirically and in a routine manner, in relation to the stated e. "he term "therapeutically effective amount" refers to an amount of an antibody or other drug effective to "trea " a disease or disorder in a subject or mammal. In some embodiments, the therapeutically effective amount of the drug can reduce the number of B-cells; reduce the number of autoreactive B—cells; decrease the symptoms of disease; or slow the progression of disease. See the definition herein of "treating". A "prophylactically effective amount" refers to an amount ive, at dosages and for periods of time necessary, to achieve the d prophylactic result. Typically but not necessarily, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically ive amount will be less than the therapeutically effective amount.

The word "label" when used herein refers to a detectable compound or ition which is conjugated directly or indirectly to the antibody so as to generate a "labeled" antibody. The label can be detectable by itself (e.g., radioisotope labels or cent labels) or, in the case of an enzymatic label, can catalyze chemical alteration of a substrate compound or composition which is detectable.

Terms such as "treating" or "treatment" or "to treat" or "alleviating" or "to alleviate" refer to therapeutic measures that cure, slow down, lessen ms of, and/or halt progression of a diagnosed pathologic condition or disorder. Thus, those in need of treatment include those already diagnosed with or suspected of having the disorder. Prophylactic or preventative measures refer to therapeutic measures that prevent and/or slow the pment of a targeted pathologic condition or disorder. Thus, those in need of prophylactic or preventative measures include those prone to have the disorder and those in whom the disorder is to be prevented. In certain embodiments, a subject is successfully ed" if the patient shows one or more of the following: decreased B-cells; decreased autoreactive s; decreased B-cell activity; decreased aberzant B-cell activity; decreased lignant B—cells, decreased non-cancerous B- cells, reduced immunoglobulin level; reduced morbidity and mortality; improvement in quality of life; or some combination of effects.

"Polynucleotide," or "nucleic acid," as used interchangeably herein, refer to polymers of nucleotides of any length, and include DNA and RNA. The nucleotides can be deoxyribonucleotides, cleotides, modified nucleotides or bases, and/or their analogs, or any ate that can be incorporated into a polymer by DNA or RNA polymerase. A polynucleotide can comprise modified nucleotides, such as methylated nucleotides and their analogs. If present, modification to the nucleotide structure can be imparted before or after assembly of the polymer. The ce of nucleotides can be interrupted by non-nucleotide components. A polynucleotide can be r modified after polymerization, such as by conjugation with a labeling ent. Other types of modifications include, for example, "caps", substitution of one or more of the naturally ing nucleotides with an analog, internucleotide modifications such as, for example, those with uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, cabamates, etc.) and with charged es (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant es, such as, for example, proteins (e.g., nucleases, toxins, antibodies, signal peptides, ply-L-lysine, etc.), those with intercalators (e.g., acridine, psoralen, etc.), those ning chelators (e.g., metals, radioactive metals, boron, oxidative metals, etc.), those containing alkylators, those with modified linkages (e.g., alpha anomeric nucleic acids, etc.), as well as fied forsns of the cleotide(s). r, any of the hydroxyl groups ordinarily present in the sugars can be replaced, for example, by phosphonate groups, phosphate groups, protected by standard protecting groups, or activated to prepare additional linkages to additional nucleotides, or can he conjugated to solid ts. The 5‘ and 3’ al 0H can be phosphorylated or substituted with amines or organic capping group moieties of from l to 20 carbon atoms. Other hydroxyls can also be derivatized to rd protecting groups. Polynnoleotides can also contain analogous forms ofribose or deoxyrihose sugars that are generally known in the art, including, for example, ‘2’~O—niethyl—, Ziflnallyl, 2'-fluoro- or 2'-azido-ribose, carbocyclic sugar analogs, .alpha.-anomeiic sugars, epimeric sugars such as arabinose, xyloses or lyxoses, pyranose sugars, furanose sugars, sedoheptuloses, acyclic analogs and abasic nucleoside analogs such as methyl riboside. One or more phosphodiester linkages can be replaced by alternative linking limited groups. These alternative linking groups include, but are not to, embodiments wherein phosphate is replaced by P(O)S ("thioate"), P(S)S ("dithioate"), "(O)NR2 ("amidate"), P(O)R, P(O)OR', CO or CH2 ("foranacetal"), in which each R or R' is ndently H or substituted or unsubstituted alkyl (1-20 C) optionally containing an ether (-—O——) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl or l. Not all linkages in a polynucleotide need be identical. The preceding description applies to all polynucleotides referred to herein, including RNA and DNA.

The term r" means a construct, which is capable of delivering, and optionally expressing, one or more gene(s) or sequence(s) of interest in a host cell. es of vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid or phage vectors, DNA or RNA expression vectors associated with cationic condensing agents, DNA or RNA expression vectors encapsulated in liposomes, and certain eukaryotic cells, such as producer cells.

The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any . The polymer can be linear or branched, it can comprise modified amino acids, and it can be interrupted by non-amino acids. The terms also ass an amino acid polymer that has been modified naturally or by intervention; for e, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other lation or modification, such as conjugation with a ng component. Also included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art. It is understood that, because the polypeptides of this invention are based upon antibodies, in certain embodiments, the polypeptides can occur as single chains or associated chains.

The terms "identical" or percent "identity" in the context of two or more nucleic acids or polypeptides, refer to two or more sequences or subsequences that are the same or have a specified percentage of nucleotides or amino acid residues that are the same, when compared and d (introducing gaps, if necessary) for m correspondence, not considering any conservative amino acid substitutions as part of the sequence ty. The percent identity can be measured using sequence comparison software or algorithms or by visual inspection. Various algorithms and software are known in the art that can be used to obtain alignments of amino acid or nucleotide sequences. One such non- limiting example of a ce alignment algorithm is the thm described in Karlin et al, 1990, Proc. Natl. Acad. Sci, 87:2264-2268, as modified in Karlin et al., 1993, Proc. Natl. Acad. Sci, 3- 5877, and incorporated into the NBLAST and XBLAST ms (Altschul et al., 1991, Nucleic Acids Res, 25:33 89-3402). In certain embodiments, Gapped BLAST can be used as bed in Altschul et al., 1997, Nucieic Acids Res. 89~3402. BLAST—2, \VLLBLASTLZL hui et al., 1996, Memoirs in Brigg/raraiogv, 266:460-430), ALIGN, ALIGNQ (Genentech, South San Francisco, California) or Megalign {:DNAS’DXR) are additional publiciy available software programs that can be used to align sequences, In n embodiments, the t identity between two nucleotide sequences is determined using the GAP matrix and a gap weight of 40, 50, 60, 70, or program in GCG software (e.g., using a NWSgapdnaCMP 90 and a length weight of l, 2, 3, 4, 5, or 6). In certain alternative embodiments, the GAP program in the GCG re package, which incorporates the algorithm of Needleman and Wunsch (J. M0]. Biol. (48):444-453 (1970)) can be used to determine the percent identity n two amino acid sequences (e.g., using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of l, 2, 3, 4, 5). Alternatively, in certain ments, the t identity between nucleotide or amino acid sequences is determined using the algorithm of Myers and Miller (CABIOS, 4:11-17 (1989)). For example, the percent identity can be determined using the ALIGN program (version 2.0) and using a PAM120 with residue table, a gap length penalty of 12 and a gap penalty of 4.

Appropriate parameters for maximal alignment by particular alignment software can be determined by one skilled in the art. In certain embodiments, the default parameters of the alignment software are used.

In certain embodiments, the percentage identity "X" of a first amino acid sequence to a second sequence amino acid is calculated as 100 x (Y/Z), where Y is the number of amino acid residues scored as identical s in the ent of the first and second sequences (as aligned by visual inspection or a particular of residues in the second ce. If the length sequence alignment program) and Z is the total number of a first sequence is longer than the second sequence, the percent identity of the first sequence to the second sequence will be longer than the percent identity of the second sequence to the first sequence.

As a non-limiting example, whether any ular polynucleotide has a certain percentage least 90% identical, and in some sequence ty (e.g., is at least 80% identical, at least 85% cal, at embodiments, at least 95%, 96%, 97%, 98%, or 99% identical) to a reference sequence can, in certain embodiments, be determined using the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, WI 53711).

Bestfit uses the local gy algorithm of Smith and Waterman, Advances in Applied atics 2: 482 489 (1981), to find the best segment of homology between two sequences. When using Bestfit or any other sequence alignment program to determine whether a particular sequence is, for instance, 95% identical to a reference sequence according to the present invention, the parameters are set such that the percentage of identity is calculated over the full length of the reference nucleotide sequence and that gaps in homology of up to 5% of the total number of nucleotides in the reference sequence are allowed.

In some ments, two nucleic acids or polypeptides of the invention are substantially identical, meaning they have at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, and in some embodiments at least 95%, 96%, 97%, 98%, 99% nucleotide or amino acid residue identity, when -23_ compared and aligned for maximum correspondence, as measured using a sequence comparison algorithm or by visual inspection. Identity can exist over a region of the sequences that is at least about 10, about , about 40-60 residues in length or any ral value therebetween, and can be over a longer region than 60-80 residues, for e, at least about 90-100 residues, and in some embodiments, the sequences are substantially identical over the full length of the sequences being compared, such as the coding region of a nucleotide sequence for example.

A "conservative amino acid substitution" is one in which one amino acid residue is replaced with another amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, irecluding basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucirge, isoleucine, proline, phenylalanine, nine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tye‘osine, phenylalanine, tryptophan, ine). For example, substitution of a phenylalanine for a tyrosine is a conservative substitution. In some embodiments, conservative substitutions in the ces of the ptides and antibodies of the invention do not abrogate the binding of the polypeptide or antibody containing the amino acid sequence, to the antigen(s), i.e., the CD37 to which the ptide or antibody binds. Methods of identifying nucleotide and amino acid conservative substitutions which do not eliminate antigen binding are well— known in the art (see, e.g., Brummell et al., Biochem. 32: 1180-1 187 (1993); Kobayashi et al. Protein Eng. 12(10):879—884 (1999); and Burks et al. Proc. Natl. Acad. Sci. USA 94:.412-417 (1997)).

As used in the present disclosure and claims, the ar forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise.

It is understood that wherever embodiments are described herein with the language "comprising," otherwise analogous ments described in terms of "consisting of" and/or "consisting essentially of’ are also provided.

The term r" as used in a phrase such as "A and/or B" herein is intended to include both "A and B," "A or B," "A," and "B." Likewise, the term "and/or" as used in a phrase such as "A, B, and/or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A ); B (alone); and C (alone). 11. CD37 binding agents The t ion provides agents that specifically bind CD37. These agents are referred to herein as "CD37 binding agents." Exemplary CD37-binding agents have been described in US.

Published Application No. 256153, which is herein incorporated by reference in its entirety.

The full-length amino acid sequences for human, macaca, and murine CD37 are known in the art and also provided herein as represented by SEQ ID NOszl—3, respectively.

Human CD3 7: CLSLIKYFLFVFNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKVL AISGIFTMGIALLGCVGALKELRCLLGLYFGMELLLFATQITLGILISTQRAQLERSLRDVVEKII'IQ KYGTNPEETAAEESWDYVQFQLRCCGWHYPQDWFQVLILRGNGSEAHRVPCSCYNLSATNDSTI LDKVILPQLSRLGHLARSRHSADICAVPAESHIYREGCAQGLQKWLHNNLISIVGICLGVGLLELG FMTLSIFLCRNLDHVYNRLAYR (SEQ ID N031) Macaca mulatta CD37: MSAQESCLSLIKYFLFVFNLFFFVILGSLIFCFGIWILIDKTSFVSFVGLAFVi’LQIWSKV LAISGVFTMGLALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRAQLERSLQDIVEKTI PEETAAEESWDYVQFQLRCCGWHSPQDWFQVLTLRGNGSEAHRVPCSCYNLSATNDS TILDKVILPQLSRLGQLARSRHSTDICAVPANSHIYREGCARSLQKWLHNNLISIVGICLGVGLLEL IFLCRNLDHVYNRLRYR (SEQ ID NO:2) Murine CD37 (NP_031671): MSAQESCLSLIKYFLFVFNLFFFVLGGLIFCFGTWILIDKTSFVSFVGLSFVPLQTWSKV LAVSGVLTMALALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRVRLERRVQELVLR TIQSYRTNPDETAAEESWDYAQFQLRCCGWQSPRDWNKAQMLKANESEEPFVPCSCYNSTATN DSTVFDKLFFSQLSRLGPRAKLRQTADICALPAKAHIYREGCAQSLQKWLHNNIISIVGICLGVGL LELGFMTLSIFLCRNLDHVYDRLARYR (SEQ ID N023) In certain embodiments, the CD37 binding agents are antibodies, immunoconjugates or polypeptides. In some embodiments, the CD37 binding agents are humanized antibodies.

In certain embodiments, the CD37-binding agents are capable of inducing complement dependent xicity, Examples of CD37-binding agents that are capable of inducing complement dependent cytotoxicy are disclosed, for e, in US. Published Application No. 2011/0256153, which is herein incorporated by reference in its entirety. For example, treatment of cells with the CD37—binding agents can result in CDC activity that reduces cell ity to less than about 80%, less than about 70%, less than about 60%, less than about 50%, less than about 40% or less than about 35% of the cell viability of untreated cells. Treatment of cells with the CD37—binding agents can also result in CDC activity that reduces cell ity to about 70-80%, about 60—70%, about 50—60%, about 40—50%, or about 30-40% of the cell Viability of ted cells. In some particular embodiments, the CD37-binding agents are capable of inducing complement ent cytotoxicity in Ramos cells.

In certain embodiments, the CD3 7-binding agents are capable of ng dy dependent cell mediated cytotoxicity (ADCC). Examples of CD-37 binding agents that are capable of inducing antibody dependent cell mediated cytotoxicity (ADCC) are disclosed, for example, in US. Published Application No. 2011/0256153, which is herein incorporated by reference in its entirety. For example, treatment of cells with the CD37-binding agents can result in ADCC activity that produces at least about %, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or at least about 60% cell lysis. Treatment of cells with the CD37- binding agents can result in ADCC ty that produces about 10-20%, about 20—30%, about 30-40%, or about 40-50% cell lysis. Treatment of cells with the CD37-binding agents can also result in ADCC activity that es about lO—50%, about 20-50%, about 30-50%, or about 40-50% cell lysis. In some particular embodiments, the CD37-binding agents are capable of inducing ADCC in Daudi, Ramos, and/or Granata—S 19 cells.

In some embodiments, the CD37—binding agents are capable of inducing apoptosis. In some embodiment, the CD37—binding agents are capable of inducing apoptosis in the absence of cross-linking agents. Examples of CD37—binding agents that are capable of ng apoptosis in vitro in the absence of a cross—linking agent are disclosed, for e, in US. hed Application No. 2011/0256153, which is herein incorporated by nce in its entirety. For example, treatment of cells with the CD37- g agents can induce apoptosis in at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or at least about 55% of cells. In some ular embodiments, the CD37-binding agents are capable of inducing apoptosis in Ramos cells and/or Raji cells.

] In some embodiments, the CD37-binding agents are capable of depleting B—cells. In some embodiments, the B-cells are autoreactive B-cells. In some ments, the B—cells are not cancer cells.

In some embodiments, the B—cells are not turner cells. In same embodiments, the B-cells are not cancerous cells, In some embodiments, the B—cells overexpress C1337. In some embodiments, the B— cells do not overexpress CD37.

Treatment of cells with CD37-binding agents can result in depletion of at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, or least about 75% of B—cells.

In some embodiments, the CD37-binding agents do not e T—cells under the same conditions in which B—cells are depleted. For e, treatment of cells with CD37—binding agents can result in depletion of less than about 20%, less than about 15%, less than about 10%, or less than about % of T-cells. In certain embodiments, the CD37-binding agents deplete at least about 25% of B—cells and e less than about 10% of T-cells. In certain embodiments, the CD37—binding agents deplete at least about 30% of B—cells and deplete less than about 5% of T—cells.

In some embodiments, the CD37-binding agents do not deplete monocytes under the same conditions in which B-cells are depleted. For example, treatment of cells with CD37-binding agents can result in depletion of less than about 20%, less than about 15%, less than about 10%, or less than about % of monocytes. In certain embodiments, the CD37-binding agents deplete at least about 25% of B-cells and deplete less than about 10% of monocytes. In certain embodiments, the CD37—binding agents deplete at least about 30% of s and deplete less than about 5% of monocytes.

In certain embodiments, immunoconjugates or other agents that specifically bind human CD37 trigger cell death via a cytotoxic agent. For example, in certain embodiments, an antibody to human CD37 is conjugated to a maytansinoid that is activated in cells expressing the CD37 by protein internalization. In certain alternative ments, the agent or antibody is not conjugated to a maytansinoid or other cytotoxic molecule.

The CD37-binding agents include CD37 antibodies such as CD37-3, CD37—12, CD37-38, CD37-50, CD37-51, CD37—56 and CD37-57 and fragments, variants and derivatives thereof. The CD37- binding agents also e CD37-binding agents that specifically bind to the same CD37 epitope as an antibody selected from the group consisting of CD37-3, CD37—12, 8, CD37—50, CD37—51, CD37- 56 and CD37-57. The CD37-binding agents also include CD37-binding agents that competitively t an antibody selected from the group consisting of CD37-3, CD37-l2, CD37-38, CD37-50, 1, CD37—56 and CD37-57.

In some ular embodiments, CD3 7-binding agents can be characterized by their ability to bind chimeric CD37 polypeptides, including murine/human and /human ic polypeptides desribed in US hed Application No. 2011/0256153, which is herein orated by reference in its entirety, and provided in the table below.

Chimeric Sequence EPoly- 3 MSAQESCLSLIKYFLFVFNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKV ;hCD37— M1 LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRVRLERRV QELVLRTIQSYRTNPDETAAEESWDYVQFQLRCCGWHYPQDWFQVLILRGNGSEAH RVPCSCYNLSATNDSTILDKVILPQLSRLGHLARSRHSADICAVPAESHIYREGCAQGL QKWLHNNLISIVGICLGVGLLELGFMTLSIFLCRNLDHVYNRLARYR (SEQ ID 5 i; i NO. 184) imuCD37-NEISTQRVRLERRVQELVLRTIQSYRTNPDETAAEESWDYAQFQLRCCGWQSPRDWNK R176 AQMLKANESEEPRVPCSCYNSTATNDSTVFDKLFFSQLSRLGPRAKLRQTADICALPA SKAHIYREGCAQSLQ(sEQID No1851 A hCD37— 3MSAQESCLSLIKYFLFVFNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKV M45 LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRAQLERSLRi 5DVVEKTIQKYGTNPEETAABESWDYVQFQLRCCGWHYPQDWQVLILRGNGSEAH ; RVPCSCYNLSATNDSTILDKVILPQLSRLGPRAKLRQTADICALPAKAHIYREGCAQS 1 LQKWLHNNLISIVGICLGVGLLELGFMTLSIFLCRNLDHVYNRLARYR (SEQ ID 1M1qu1861 M 5 hCD37m MSAQESCLSLIKYFLFVFNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKV ECD- LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRVRLERRV H45 QELVLRTIQSYRTNPDETAAEESWDYAQFQLRCCGWQSPRDWNKAQMLKANESEEP RVPCSCYNSTATNDSTVFDKLFFSQLSRLGHLARSRHSADICAVPAESHIYREGCAQG 1 LQKWHNNLI$1vGICLGVGLLELGFMTLSIFLCRNLDHVYNRLARYR (SEQ ID N0: = s 1871A AA hCD37m MSAQESCLSLIKYFLFVNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKV ; H5 LAISGIFTVIGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRVRLERth ~27- W;—IQVELVLRTIQSYRTNPDETAAEESWDYAQFQLRCCGWQSPRDWNKAQMLKANESEEP RVPCSCYNSTATNDSTVFDKLFFSQLSRLGPRAKLRQTADICAVPAESHIY’REGCAQG LQKWLHNNLISIVGICLGVGLLELG’FMTLSIFLCRNLDHVYNRLARYR (SEQ ID NO: ;188) ’ hCD37H";MsAQESCLSLIKYFLFVFNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKV ECD-H4 LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRVRLERRV s QELVLRTIQSYRTNPDETAAEESWDYAQFQLRCCGWQSPRDWNKAQMLKANESEEPCE RVPCSCYNSTATNDSTVFDKLFFSQLSRLGHLARSRHSADICALPAKAHIYREGCAQS LQKWLHNNLISIVGICLGVGLLELGFMTLSIFLCRNLDHVYNRLARYR (SEQ ID NO: ;189) 'iiémné‘i‘lwé"MsAQESCLSLIKYFLFVFNLFFFVLWIWILIDKTSFVSFVGLAFVPLQIWSKV Mac4 LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRAQLERSLR: DVVEKTIQKYGTNPEETAAEESWDYVQFQLRCCGWHYPQDWFQVLILRGNGSEAH YNLSATNDSTILDKVILPQLSRLGQLARSRHSTDICAVPAESHIYREGCAQGL QKWLHNNLISIVGICLGVGLLELGFMTLSIFLCRNLDHVYNRLARYR (SEQ ID NO: M3045 LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRAQLERSLR; DVVEKTIQKYGTNPEETAAEESWDYVQFQLRCCGWHYPQDWFQVLILRGNGSEAH 5 '5RVPCSCYNLSATNDSTILDKVILPQLSRLGQLARSRHSTDICAVPANSHIYREGCARSLj hCD37— ;MSAQESCLSLIKYFLFVFNLFFFVLGSLIFCFGIWILIDKTSFVSFVGLAFVPLQIWSKV MacS LAISGIFTMGIALLGCVGALKELRCLLGLYFGMLLLLFATQITLGILISTQRAQLERSLR.

;DVVEKTIQKYGTNPEETAAEESWDYVQFQLRCCGWHYPQDWFQVLILRGNGSEAH ; RVPCSCYNLSATNDSTILDKVILPQLSRLGHLARSRHSADICAVPANSHIYREGCARSL; QKWLHNNLISIVGICLGVGLLELGFMTLSIFLCRNLDHVYNRLARYR (SEQ ID NO: ' 192) In some particular embodiments, the binding of the CD37-binding agents to CD37 does not require human CD37 amino acids 109-138. Thus, some CD37-binding agents bind to a polypeptide comprising the amino acid ce of SEQ ID . In other embodiments, the binding of the CD37— binding agents to CD37 is ted by mutation of human CD37 amino acids 3. Thus, some CD37—binding agents do not bind to a polypeptide comprising the amino acid sequence of SEQ ID NO:185.

In some embodiments, the CD37-binding agents bind to a polypeptide of SEQ ID NO:184 and to a polypeptide of SEQ ID NO:186, but do not bind to a polypeptide of SEQ ID NO:185.

In some embodiments, the CD37—binding agents bind to a polypeptide of SEQ ID NO: I 87. In some embodiments, the inding agents bind to a polypeptide of SEQ ID NO:187 and a polypeptide of SEQ ID NO:l88. In some embodiments, the CD37—binding agents bind to a polypeptide of SEQ ID NO:187 and a polypeptide of SEQ ID NO:1 89.

In some embodiments, the CD37-binding agent binds to a polypeptide of SEQ ID NOzl90, but does not bind to a polypeptide of SEQ ID . In some embodiments, the CD37—binding agent binds to a polypeptide of SEQ ID N02192, but does not bind to a polypeptide of SEQ ID NOzl9l. —28- {1311119} CD37 peptide fragments to which certain inding agents bind to include, but are not 1111111611110, CD37 fragments cmnprising, censisting essentiaily Of, 01 consisting ot‘aminc acids 2004243 Qt" SEQ 173 NO: 1 amino acids 2213 or SEQ H3 NO:1, or amino acids 2216343 of SEQ 1'13 N131. in some en1b0<11nen1‘» the L133Lbinding agent is 1cal£y binds to :21 1111111311 C1337 epitape (30111151131111: amino acids 2029.43 of SEQ ‘13:? N13], In some embodiments, the binding, 11fthe C1133 7—b111d1ng agent 10 CD37 requires 3.1711130 acids 3 of SEQ ID ‘NO:1 1n scme embodiments, the binding 01': the (31337,- 11111111119, agen1 111 CD37 requires amine acids 2013—2213 0.1" SEQ 1D 710:1. In some e1nbnd1n1en1s, the binding 11tts1e CD37~binding aoent to (31133 7 requires 1111111115 acids 221-243 nf SEQ ID NO:1. 11331213} Exampies of CD37~b1ndit1gagents with the afctementioned binding prnperties are desctibed in US. E11blisned Application No. 201 170256153, which1s herein incorpersted by reference111 11s ty. 113131211 The C1D37~b1nding agents also finciude (SEW—binding events that comprise the heavy and 119,111 chain CDR sequences of CD373, 131337—12, {1337,68 CD37—513,C1‘337«51 €1337~56 or {7133757, 71113111131157 and fight chain CDRS of CD3 7~3 8, {3337513, 151337—51, 66 and CD576? contain related sequences T11e1efcre, the €133 7—111111111157, agents can 111515 comprise heavy and light chain CDR sequences that cemprise a consensus sequence ed by the alignment of 37~58, (3.133760 CD3751, CD37~ 56 and —57. The CDR sequences 373, (1337—12, (311337—38, 513 CD 75'1, {71337-56L1.) and C133",57 as welt as 11.16 cnnsensus sequence of U33733, CD3750 (£337—51, 131337-56 and C1337 57 ate descfibed 1n Tatfies 1 and 2 beiow.

Table 1: Variable heavy chain CDR amino acid sequences " """ Anubod1 3 .111611111 1 412.11... VHCDR3 (:‘153"7"-3 TEG‘VQ (6161511?V‘1‘1VQ‘DQS'v1N1SEQ1D GGYSLAH ("SEQ 115NO"-‘6) CD37—12‘"E EYQVN{$131335 WINRTQEQRRLQ133"" G'I‘VVAD (SEQ 11'.) N09) N37) 138) i 1CD3735‘11"""""""""§ Sewnvmsmtn 111176111111116113111 GYYGYGAWFVY(SEQID N0.133). \O11} ,,,,, No 12) CD37—50 5 61117111111 (SEQ 1D 1511611611'1611111‘1‘5" AWFAY (SEQ 115 : NQ131 \Q1713 :=NQ-15_)_____ CD3751 "s"1‘.5156151711 (SEQ1D""571111?8(15‘1‘11(61715515GYYGFGAWFVY "(‘S‘E‘Q1‘15"" NO:16) \Q1") N018) CD3756W1(SEQ 1D GYYGFGAWFAY(SEQn5" M N0"2.1.3 ..

CD37-57 S15EAw1-11s1-1Q1D AWFAY (SEQ ID CONSENSUS SQEtA or 11 1 _ .61 1111‘ S1111} 01‘ §GYYG[Y or FjGAWF[V or {SEQ113 NQ25) gsn‘[DV01" N} (SEQ 11.1 :AgY. (SEQ ID NO27) MMLNQ;.161 ..... 3 252-31 ESYGMS (SEQ ID SSQQSHYSEDWEQ .HSYYDTSVDY NQ171). 1.1 .. _‘nn.......1 _ ;__(__S__E_Q_IQNQ173) 252-3 SYQMS (SEQ ID 1 1 HSYYDTSVDY (SEQ ID Table 2: le light chain CDR amino acid sequences WV1.CDR] VLCD" ':'yLCDR3 CD373 AS1N1RS1S1A'STQ VATNLAD 2SEQ 2D QHYWGTTWT ('SEQiD"" 2 12321:281 _ENo30E - NQ2.9.). 77777777 CD3712 NSTSSSSYLY 3 YA‘WLA‘S 2S1Q 113 QHSWEIPYT D" ..............................................E-CD37 ESEQ113NQ11E 2222222 "MEN033) 38 5 :13 ._ TYMH (SEQ , E QQWISN'PET(SEQ'1'D"" g 11') N0:34) QE3sE §_E _.1 1CD37-50"""'""" SATSSVTS121112SLQ12"'.2<21212 2STQ 123'"WE PPT (SEQ ID ' 44'4th 19% ,, 1Huma11ized ;1.11SN112v 2SEQ 1D 3.1.2.111 ,,,,,, "W1 __m__w ' CD37-51 SAT‘SSVT'VWQ‘SEQDTSKLLYS1SEQ 113 1DN042 ND4") ii"QQWSSNPPT (SEQ IDN043_)_mw 1CD37-56 ESASSSVYMHESTQ EDTSKLS(.. QQWISDPPT(SEQID ; Humanw‘d DTSNEAS 2SEQ1D N1317); wa. _M.. EEEEE CD37—57 SA1SS\ITY\1112SEQ D2SSLLSS 1S1Q113 E NPPT (SEQ 1D E EDNDESLDMD ND49.}. M...................................................., 1121111aniz'ed DT‘SNxL‘18 (SEQ ED CONSENSUS SA'"['T'"Qr S]SSVTYMHDTSaK S} N''E'L'E'11611211 'QQWU or S][S or D][N or (SEQIDNEQ:52} 9.4 YE("SEQ 1QNQ: 513 DPPT (SEQ IQNQ_:SE41 252-3 RASQD1SNYLN(SEQE"1'TSSL12S2SEQ'1D QQGNALPWT(SEQ 1D" IDNO.174.).ANDY/'3)No 176) The CD37 binding molecules can be antibodies or antigen binding fragments that specifically bind to CD37 that comprise the CDRS of CD37-3, 2, CD37—50, CD37—51, CD37—56, or CD37—57 with up to four (i.e., 0, l, 2, 3, or 4) conservative amino acid substitutions per CDR.

The CD37 binding molecules can se one of the individual variable light chains or variable heavy chains described herein. Antibodies and polypeptides can also comprise both a variable light chain and a variable heavy chain. The variable light chain and le heavy chain sequences of murine, chimeric, and humanized CD37-3, CD37-12, CD37-50, CD37-51, CD37-56, and CD37-57 antibodies are provided in Tables 3 and 4 below.

Table 3: Variable heavy" chain amino acid sequences Antlbody WAmmo ':':Q 113 N13) ......... muCD37-3 Q‘vQ‘s!KESGPGT1‘17APQQSLQKl ‘v 41111128131 lSQVS‘WVRQT’PGKGLEWLGVIW wUDUS1NYHMDIKKDHNNSOVTLixLN‘SlEQETE'E13E}1EA'EYYCAkGiEsE‘Y(3142......

E 11WGQG 8.8I888A{S1Q1DN1}888 E E QVQVNLNAAA'17A8I8Q88 bITLT‘v8G8'8'1I1'1SGVSWVNOPEMKGI8‘88; .

E GDGSTNY1-1SALKSR1.8118.181AHSKSQ‘VFLKLVNLQ11.313.1‘15‘81‘88CAKGGY’8.8.A .88.) NQ.5111 -1 huCD3WE 111181101511VTV11183111) '1: E GGG88N8I88888888888888.88888828SQVFLKLN88..LAAG8‘A8‘8-YCANGGY‘88...AQVQVQESGWN7Y’APSQT1.S11CTVNGFSL188GV888I8I8Q8>8>8'88:.G888888G8I'8'18I'.‘ E 8888GQG888/88/88(61,011) N08 II) huCD373v1 .1 EQIvG8/Q838GPGL8IAP8Q8 I S1‘1CTV‘818FSL1TSG‘VSWVRQPFGMN11888(88888..

E GIGGSTNY8888888888888.888.*HSKSOVFLKENSLTAADI8‘A‘I8‘88I8:AKGGY81A E HYVGQGHV8888 8N8}588 muCD3712 E QIOLV’Q‘GPELKKE‘GE8828888.88>888883882888888:88II8G1I'8'8I1818888888888888888888188888888-88 11YSGG1D‘Y‘NPS1KSR1S11RDTNK‘NQFFLRLS‘8V188-888A8‘8Y1A81888IG8GE.

E__A88-88I888GQG88.8I8-8Ag88Q188NG:§88_ .....

Q‘v’QLQ'E8888888.8IKPSQSLSLTLTV1GY'8.818888888888888881288888888888811'8" ILY’SGG11‘)Y\1">1KSR1S1TRDTSK‘NQ1‘11R1SSV1"11313181381"YY1-ARGYYGYG AWFY’YW '7 "1‘\ISA (SFQ 111N13: 137) 1 hu7CD37-38 "1"QVQ8Q888888.Y1.888883888888.8‘8I8G8 88'8187181181188888I888Q88A88888888888888 LYSGG{DYNPS}.RS181SURETSKNQFFLRLSSVILYDiA1YY1‘ARGYYGYG AWFY’YWGQG1L "7 ‘ 771117111313'1'7'1'50 D‘sQLQESNPDH' ~ . 1YSGST‘VYSPSLKSRlS11'RD1'SKNHFTLQL‘NSV111313.1'A1'YY’1‘A1118YY(1Y1: 1.A.WEAYYVGQGTLYTVA (SEQ 1.1.) N173:E64} . huCD3750 ESAJ’GLLRRSQ‘AS1":11Y7‘VSGYSITSG1AWH‘MRQHPGRKE..... .8 1.1.YSGST‘vY’SPSLKSRIS11'RDTSKNHEF1.Q1.NSVTAADTATYY"CARGY’YGYG ANY/1*AYW1NY1zTLYTVSAgShQ11}1‘8065) """"""""""" huCD3751 .. QIVFSGPEVLKPGENLS11‘13 ..1.8Y"81">SG1"YYY11YY’1R1‘1PGRGL}?WMGY1 HYSCxSTNYbPSLQGRISI111388118131418G1..N88I8A8838AI8888ARGYYGEGA E8888I8I88IGQ88888888188A 88818:: 888 No67 E muCD3756 ESGPDLVKPSOSES1.'8'G'I'8'I'1I"8G8I88I8‘8C8A88I888888Q8~1"(J\K18 88-88888 E (SiAW‘8.A8EW880G888-11‘"8A888G.11.} N88 88888888SQGTNY’TNPSLKSRVS11111.71.SNNQ811'.QLNS‘VTTEDTATYYCARGYYGF - huCD3756 QESUFGLY1§PSOS1S1.":11'1VS18YS1T‘8181'A‘818’11W1RQF"GKG183.88888888 HYSGGTNYNPSLKSRV81111111818111":8‘8.Q8.NS‘JTAADTATY’Y 8::ARGY’YG8‘ GAVE-8A8 88888888888888A ‘88 111180 889) muCD3757E8'8VQLQJ}.8G883888888888888'8..I8I8i'8I17I11YYS1TS8LY1AWHWIRQE88888888:W7881888 E LY 81718.17VYSPSIKSR1S1TRD1'S18LNQ1‘1IL01.NSV17'1'11131TATYY1,I‘YRGYYGYG :- Y‘VE‘AYWGOGNV1VSA8SFQ1DN1)I111" huCD3'1'l75'7""""""""' QVQLQESGP1Y181.K1‘SQSISLTC1‘11‘8113/ .‘311‘8G1IAW11‘8‘8’1RQF131EK(}';1.FKY’MGYI' LYSGSFIS‘JY13881811SE18.1S1'1R1)TS18NQFF1:QLNSVFAAD11A1'YY’1.‘.ARGYY(1‘11: EVQ‘V 777712717. . .77 A81sFTFSSY1x‘81S‘V‘v'YRQ11’DKR11YV’VA11 SSGGSYTYSPDSY181811.?11‘8181)\AKK1'1Y1.Q1\/1SS1.KSED1AMYYCARBSYY’ Table 4: Variable light chain amino acid sequences lbOdyWM"\I’L""AAI1')AAEAI1Q "’"Q {KYLE EL") NA?) :_.. muCD373 DIQVLIQENPAQLNV‘NV’TTTYT:TLTLANTNTTTNNLAYYIYQQKQLTNNTQLLYINYAT NLADNYITNTLTNLTNTTNTTTQYSLKINNLQNTTTTLTTYYCQTTYYYITTTTYYITTTTLTQTN 1...... chCD°73 .. QT;TYTLTLTLANTNTNNNTATTYQQKQLTNNTQLLYINYAT LTYTNNTNLTNTTNT:TTQYNLKLNNLQNTTYTQTYYCQTTYYYITTTTYTTTTTLTQTN: LTTKRTNLQ LQAAANQ-73) AAA: huCD37—3 LTTQYLTQNTN T..NYNYILTTNYITLTCLANTN . _ ANYQQNYLTLNTNLTYINY‘AT T (1.0 and 1.1) NELADLTYITNNT‘NLTNTTNQTQYNLNTNNLQTTgTzTTQTYYCQTTYYYITTTTYYTTNQNTN LEEKRT‘EQELEM E -12ED1‘Y’}TQNTANTAYINTTTQNATTNL‘NANQN 'NYNYLYYY TALTQQT E EYANNT.ANLTYITEANTNLTNQNQTQTTLNTTLTTY TNT-.TDTATYYCQTTSWTLPYTTQLTG E TLLLTLTTAATNTIQ TATAATAAAAANT}755} chcfiéiié""""""""""""""""ELTTYI‘TTQNTAN‘LAYNLLTQTLATTNT‘TTANQNYI‘NTNSYNYLYYYTQQLTNQTTKLTTN EYANNT.ANGY‘TARTSGSGSQLQTTLNTTLTVTTLTTTATYYLQLTNYTLLTYTTQL‘TQ FEYEIEEEYR E"TE"? E E? "NEE? 75E".

L.E... muCD37—38 E QTYLTQNTATNTNAN‘TTTLLYITNT'T‘CNAsSNY’TYNTTTYYI’YQQNNLTTNTNNYYI'TY'DTS LLANQYITATTFNQQQNLTTNYNLTTSNMTJATTQAATY’YCQQYYTNN‘TTTT‘QQQTTLL LTLTHNTQ ED N012?) 77777 chCD37—3 8 QLYILTQNTTIATMNANTILTLLY"TNT"TLNANNNVTYYITTTYYYQQNNTTLNTLLYYITYDLs ENLYSCTVPARLENLTGG‘YLTTYYSL1155MAFL").AATY‘1CQQWLSNPPFF(EGGEKL TTLLT {NLQATAQAAANQ TY) huCD37—3 8 ""E"DLYLLTQNTANYTNANPQLNYITYL"T"L"N"ANNNY’TYNITTTTYYYQQNTTTTNTLTTYYITYTT"T"N"" E ENLANQYPARTNLTNTTNLTTNYNLTLNNNLLANDAATYYTQQTNITNNTTTTQQGTNL E:TALLAN TNLQ ATAADA NO-21"AAAA muCD3T7—50 SEQ":Y""L"TQNTIALYTNA PC ,LYTYLTLNEALNNYI'LYYTTTYTYQQNNTTTNTKTTYYILY"DL"N" E EKLPYQYNTNTNLTNLTNLTTSYSLTTNNNTEAEDAATYYT:QQYYNDNTTTTGNLTTLL T:LKAARAA ATNLQ ATAQ NQ..AEEE...

E huCD37—50 TLYLTQSPLY"L"MNANTQTLYTNTTT:NATNNYIL‘T’VEHWYQQLTCTQNTNTTWLYLIL"N" E ENLTYQYTANTNQSLTNL‘TTNYNLTISSMEAEDAATYYT‘QQYYINTTNTTTTQQLTTLL NLTNTQTQNQNT) E huCD37-51..

TANTNTTNLTNITLNYNLTTNNMLIALLTAATYYCQQYYNNNPTITTLTQNTNL_ E (NLQ ALAQ NAQ:ANATATA AAE QE:TANTT ‘ muCD37-56 "QTY"L"LQNTALYTNANTTTDNYTMTLNANNNY"TY, . 'QQNNGTNTNRWLYLTTN E EKLASQYPALLNGQL‘TNLTLNY3LTTNTMEALQAATYY’CQQWLNQTTITTLTLTLTTKL AAAAAQTTNAATT(NTQ LQNE}843 AA huCD37—56 QTYLTQNTATNLNANTLTTLYILMTT:NANNNYTYNTLTYYIYQQNTTTQNTNRYYLYQTN ANQYI’TNTTTNTTNTTNQLDY.NLTTNNMT;ALDAATYYCQQWTNQTTTTT'TQTTTKL LLNN TNTQ LT)NQEAAA85A} WANT/E.")"‘~le ‘~"}("/15 NTIATYLNANTLTLKYIINLET.NA"LNNvYMTLYYYQQNNLT"T"NTNNYYITYEST""N" E TALTTNTTNTTNGTNY'NLTLNNYTLANLYAATYYLQQYY’NQNTTTTQNQTLLE FEAKARAA ENTQ TQ NQNAAAA huCD37_5Z.::""" ________________________________ PARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSDNPPTFGQGTKL MINEIKR(SEQ ID NO:872 7777777 252—3 DIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTS i 3KLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNALPWTFGGGTKL ELISEQEQ112N9111§>m \\"\V_‘."I....u.uu.........w......_______...r.r.\\ Also provided are polypeptides that comprise: (a) a ptide having at least about 90% at least about 90% sequence identity to SEQ ID NOsz55—7l or 177; and/or (b) a polypeptide having In certain ments, the polypeptide ses a sequence identity to SEQ ID NOs:72-87 or 178. polypeptide having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NOS:55-87, 177, or 178. Thus, in cettain embodiments, the polypeptide comprises (a) a polypeptide having at least about 95% sequence identity to SEQ ID NOsz55— 71 or 177, and/or (b) a polypeptide having at least about 95% sequence identity to SEQ ID NOsz72-87 or 178. In n ments, the polypeptide comprises (a) a polypeptide having the amino acid sequence of SEQ ID NOs:55-71 or 177; and/or (b) a polypeptide having the amino acid sequence of SEQ ID NOs:72-87 or 178. In n embodiments, the polypeptide is an antibody and/or the polypeptide specifically binds CD37. In certain embodiments, the polypeptide is a murine, chimeric, or humanized antibody that cally binds CD37. In certain embodiments, the polypeptide having a certain percentage of sequence identity to SEQ ID NOss55—87, 177, or 178 differs from SEQ ID NOsz55-87 by conservative amino acid substitutions only.

Polypeptides can comprise one of the dual light chains or heavy chains described herein. dies and polypeptides can also comprise both a light chain and a heavy chain. The light chain and variable chain sequences of murine, chimeric, and humanized CD37—3, CD37—12, CD37-50, CD37-51, CD37-56, and CD3 7—57 antibodies are provided in Tables 5 and 6 below.

Table 5: Full-length heavy chain amino acid sequences "but.............,,."______ Ant1bodx \ FullLenvth Heav» Chain AmmoAcidSequence(SEQ IDNO) \ ,,muCD37—3 i QVQVKESGPGLVAPSQSLSITC‘VSGFSLTTSGVSWVRQPPGKGLEWLGVIW : GDGSTNYHSALKSRLSIKKDHSKSQVFLKLNSLQTDDTATYYCAKGGYSLA HWGQGTLVTVSAAKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTL LSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTK VDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVV . DVSEDDPDVQISWFVNNVEVHTAQTQTEREDYNSTLRVVSALPIQHQDWM SGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLT CMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKN WVERNSYSCSVVHEGLHNHHTTKSFSRTPGK (SEQ IDNO:88) § chCD37—3 QVQVKESGPGLVAPSQSLSITCTVSGFSLTTSGVSWVRQPPGKGLEWLGVIW YHSALKSRLSIKKDHSKSQVFLKLNSLQTDDTATYYCAKGGYSLA HWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS EWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT E KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV E WDVSHEDPEVKFNWWDGVEVHNAKTKPREEQYNSTYRWSVLTVLHL DVVTNETKEYKTTYVSNKATPAT’TTKTTSKAKT:QT’TETT’QVYTT.TTESRDETTKNQ T VSTTCT.VKCYTY’PSDTAVEVV’T‘S\TYQETNNYKTTT’T‘VLDSDQSTTLY’SKTTVDKT ‘ STST’EIK YSTQ TD NQ:S9) T I"Hfiéfié'i'i'fiV‘TBT QVQV’. TTSLYVSVVVKQPEUKLYTEVVTUVTVV . VSGEST , . T GTTGSTNYTTPSLKSRLSTKKDTTSKSQV4TKT.NSTTT\ADTATYYCAKEEYYSEA HVVT.IQEYTTVTVSSIVSTKGESVTFEAT’SSKSTSGQTAATETCLVKDYiPEPVTVS ALTSGVHTT‘T)AVEQSSLYLYSLSSVVTVT’SSST..(YT.Q.TYTCNVNTTKPSNT 3 KVETKKVEEKSCDKTHTEPECEAPELEGT‘YT’SVELEET’KT’KT)TT.TVTTSRTEEVTCVT V’TJVSTTEDT’LVKENVV’YVTTTJVTVTT’VAKTKPKFEQYNSTYRVV-SVLTVT.HQ T TNT/"LNGKEYKCKVSNKATPI‘TT’TEK'T‘TSKxKETQPRET’QVYTLT’PSRDELTKNQ VST.TCEVKGTYT’SETTAVEVVTLSNTJQEENNYKTTPEVTTTSDQSFEEYSKLTVBK SKVVQQTYNVESESVTYTTTT‘ALTTNTTY’TQKSLSTSEQ {SEQIT)NO99‘ at" huCD373V11 ESGEGLVAPSQ"STTSTTETVSGTSLTTSGVSVVIRQT’EQKEYLEWTTIVTW g ESTNYT'TSSTKSKLSTKKDTTSKSQVE’T.KTNSTTAADTATYYCAKCTGYSLA HVVGQG’T‘LVTVSSASTKGPSVET’EAT’SSKS'T'SGG'T‘AAT..GCT.VKT)YEPEPV'T‘VS VTI'NSGAL'T'SC:V'TTTEE‘AVLQSSGLYSLSSVVTVESSSLCY'TQ'T‘YTCNVNTTK?SNT KV'DKKVF.T’KSCTBKTTT‘TCPPCPAPELEGGESVETEPT’KEKTT'T'EMTSRTETTVTCV VVDVSHEDEEVKENWYVDGVEVITTNAKTKEKEEQY’NS’T'YKVV’SV'LTVETITQ DVVLNGKEYKCKVSNKATEAE‘TEKTTSKAKGQT’RE.T’QVY"TT.T)T’SKTJETXTKNQ VSTTCLVKGEYT’STTTAVEWESNGQE‘FNNYKTTT’DVLDSETGSE‘ELYSKTXT‘VUK SRVVIQQGNVITSLSVVTTTEATTTTTHYTQKSTTSTST’G TST‘Q TD NQ19.1.). muCD3712 QTQLVQSTYPETKKETTE.TVKTSUYASLYYTTTKY’EyVTNVvVKQl—YQEIKTYTKW‘YTT: WTNTNTTJESKNAEEEKETKEAEST.ETSASTAYTQTNNEKYEDTATYEEGTEGTV V’,ADVVGQLYTTLTVSSAKTTAT’SVYET.AEVC{.YDTTGSSV’TEGCLVKGY’EPET-TV ’T‘T'T’VVNSGSESSGVTTTET’AVEQSDTY'TLSSSV'T‘V‘TSSTVVI’T’SQSTTCNVATTT‘ASS TKVDKKTET’RGPTTKEE,T"PCKCPAE\LLUUESVETTT’T’KTKTYVTTVITTST.SPTVTCV VVDVSEDDT’TTVIQTSWEVINNV‘3VITTTAQTQTTTREDYNSTLKVVSATPTQHQT) VVJVITSTsKEEKCKVNNKDLT’AETERTTSKEKSV’RAPQVIYVLT’T’T’EEETVIT'T'KKQV TTTC.IVTVTDETVTEEDTYVEVV’TNNGKTET..NYKNTTPVTDSDQSYEMYSKLKVE KKNVYIVERNSYSCSVVTTETYLTDTTTTT’TT ‘YanguK (SEQ TDNQ92) __ __ TVEEEYYI12 "I \ SGPETKKPGETVKTKEKASETY ’T Y T‘ "'YQMNTYIYKQAQQKQLKYYYKT WINTNT(YTSRNATEYKGKFA}SLEY8As.TAYY.QTNNTKYEDTIYTYECGRGTV V'ADVVI’GQG’T‘TTJT‘VSSASTKGPSVFPLAPSSKS’T‘SGG’T‘.IV.AT.GCT..-VKTTYFPTEPVT E YISYYNSQALY‘KGYH‘T‘YPAVITQSSQLYSLSSWYYPSSSLQY'QYYTCNYNHKPS K’YKYDKKYEYKSCDKYH‘TCPPCYIAYELLQGYSVKLKPPKPKQYLMTSK'TYIKYI'Y" g CV’VVDVST-TEDPEVKT‘NWY’V’TXTVI‘EVHNAKTKPREEQY’NSTYRV’VSVL’T‘VL HQY‘YWTNQKEYKCKYSNKALKAYYKKYYSKAKQQYKEPQYYYLPYSKDELYK = NQVST.Y‘QLYK"TYPSDTAVI’TZWESNGQTTE‘NNYK‘T‘TPPVTDSTTGSFFLYSKLTV ' YYIQK (SEQ m NQ:93Y ‘ T KSTYVV’QQTYNYIESESV‘YTTTTZALTTNTTYT‘O I ‘ I: muCD3738 SGPDLVIKPSQSLSLTETVTTYYSTT QYYHYYIITRQFPGNKLEVV’MAY’ ; TYYSGETTDYNPSLKSRTSTTEDTSKNQFTI‘T.KY.SSVTTTLT)TATYY’CARGY’YGYG AYTIKVYIWGQQ‘T‘LYTYISAAKYYPPSYYYITAPGSAAQ'T'NSKITY'I'LQCLYKQY‘YY EEVI’T‘VI’T‘WNSGSTSS"G‘VI'TTTFEAV‘LESDLY’TLSSSV’T‘V’PSSMKPSE’T‘V‘TCNVAH 13ASSTKVDKKTVPRDCGCKPCTC’T‘V’PEVSSVETEPPKPKDV’LT‘T’T‘L’TT’KVI’CTCV‘V’ EVI’TJTSKTTDPEVQFSVVI’FV’DDV’E‘VI‘HTAQ'T‘QPRE.EQFNSTFRSVSELPTMHQDWL I NGKET’KTKYINSIYAFT‘APTEK'T‘TQK’T‘KGTTT’KVT’QV’Y'T'TPPPKTQMAKDKVSLT CMYTDFTPESTTV'EwQYYNQQYIAENYKNTQPTTVITNTNGSYTVIY’SKTNVQKSN V‘vEAEYSTFTESVETTETYLTTNTTTTTEKSTAHSETYK SEQ TD KY ' chCD3738 OVQLOESTJPDLVKESQSTST...TCTVTTYYSTTSEYF YTYIHWTKQ KLEWMAY T TE.YYQGTIYYSPYLKYKTYYYKDTSK\QEETKT.SSVTTEDTATY’YCARGY'YGYG AWEYIYIYVGQQ'IYYTVSAASTKGPSVT‘PLAPSSKS’T‘SGGTAATQC}V’KTJYEPE T PVTY’SVV’NSTYALTSGVHTEPAVTQSSGLYSTSSYYYYPSSSTQTQ":YY’YCK-VNH KESNTKVT‘PI‘N:T j____1_g§g§§f_ .. APETTWETGPSVFTYYYKPKDYTMTYKYYT .....

VTHQTJWLNGKEYKCKVSNKALEA1‘1EKT‘1SKAKGQPRE1’QV‘1’1‘LEE‘SKI‘Hr‘iT3 TLNQVS1111"LVKGT‘YPSDTAVEWE‘SNGQEE\‘VTYKT1E‘E‘VLDSDIGS1‘1‘L‘1SKL1 V’DKSRWQQCNV’ESCS‘s]V1111‘1ALTTN11"!1‘QKSES1ST‘G (SEQ113N1):9§1 3huCD3‘7l‘é‘K‘""""""""""1Q‘1"Q111ESGEGTVKE‘SQSLSLTC1V’SGYST.TS(‘1E‘11111’}TW1111'TT‘PGKLzLE‘v‘TI'\LAY :1‘DYNEST.KSRTSTTRDTSKNQEE14111.SSV’1AA1)1‘AT‘1YCARGYYCYG 1A\VEVYVV’GQGTL‘S’‘T‘VS SAS‘T‘KGE‘SVFELAPSSKS'T‘SGG'TQAA1.11311‘1LV’KT3‘1’E1‘E EV‘TV’SWNSGAL'T‘SGV"ET"1.‘.E‘T3AV’LQS 1"SLSSV’V"T‘VT‘SS Q'T‘Y‘1‘CN‘VI‘N11 K1)SN'T‘KV’TDKKVEPKSCDK‘T‘ETTC1‘1‘1‘3PAETELLGQ E’SVELET’EKT‘KDTLMTSRT‘1’ EV’1‘CV‘1’V11V’SE1EDPEVK1‘NV‘1’YV’DL‘EV’EVHNAKTKEKET'QYNSTYRV"‘1’V’LT I’LE1Q11V‘1I‘T.1N1xKE‘YKCKV’SNKALEAE’TEK'118K1AK1.‘TQEKET—‘QV'YTLEESRDEL1‘1 KNQV’SL'1‘1‘1‘1’K111"YESDTA‘V’EWESNQQEE\VYK1‘11’11‘11.41‘1S1K3'STELYSKLT muCDE760 L‘VQLQ " ‘111’111TKESQSLSLTCTV’TCYS1‘1‘SUEAVYTT‘WTRQEPLTWLEWMGYT LYSGST‘VI‘1’".1‘81.KSRTSTTKDTSK\111ELQLNSV"1‘TEDT1A'T‘Y‘1CARGYYG‘1Cw AWE1AYV‘1’{JQU"..1TV’1"1’SAAKTTAE‘SV’YPLAPV’C.(111'1‘TGSSVT1.111ET1V’K13YT‘T‘ ii EEV‘TLTWNSUMSSGV’H.1EEAV’TQSDLYTLSSSV'1V"1SST‘VVIT’SQS1TCNVAHP 1 ASSTTLVDKKTEEKGT’.T.1K1‘CEECKCEAT—‘NLLGGPSVE‘TE‘ET‘KTKDVLMTSLST—‘TV 'T‘CV’V‘VDV"SEDDPDV’QESVVEV’NN‘1’EV’ETTAQTQTELREDY’NS‘T‘LKVVSALT‘TQH . KEFKCKVNNKDLKALITKKKSKKKGSVKAPQI1I1'VLEPPEKKM'LKK QV’111.11?\L'V'LDCMPKCIYVLWIINNCKTKL1NYKNTE1T‘V‘TDSDGSYFMYSKER AILKLKWWKNWSCKyyTKI‘LKKLKLL T‘KSFSR131.111031)NQ;9. huCD3750 QVQTQESGT‘GLLKPSQSLSLILLI115111‘8.11811?AVVHWRQTLLIUNKL13';1111111 1 1L1SCS1‘11‘SPSTKSKISN113181911113310!.NSV’TAADTATYYCARGYYGY<3 .AVV’FA.Y"1‘1’<.TQGT1VTVSAASTKGPSVE’PLAFSSKST8661".AALCCLV’KDYEPE; PVTVSWNSGALTSGVH"1‘T3T‘1A‘1"1.1QSSGLYSLSSV’V"E‘VI’PS831.1(1’T‘Q’1'Y‘TCN‘JNH 1 1 KPSNTKVDKKVIEPKSL311411111PPCPAPELLGGPSVELE1’PKEKD1.1:VETSRTP 1'V’11‘1I‘ v1131'STTFDPEV’KENWYV’DGV’EVHSIAKTKPKLLQWK11'RVVSV1"11 VL11QDW'1.NCKLYKLKVSNKALPAPTEK1‘18KAKLzQE‘KFPQVYTLPFSRDLLr T KNQVSL101.V"K1’.T1~‘YT‘S1C)1AV’E\VEQSCQPLVNYKT.TPPV’T.DSDQSE‘EL‘YSKLT VDKSRVVOQLJ\ " NHYTQKSLSLSP(SEQ11) NO9K) . .

I. . muCD3751 DVQLCLKLTKDL ' I GE‘A‘v‘TI1T1YTRQT‘E1.rNKT.E‘1‘vV11111" . .

HYSGSTNYSESLKSRTSITK1>SSK\QFELQL1\SV11ED1A1YYP1'.)VQ1SWFVNNV’EV’ET’1‘AQ'1‘Q'T‘TTREJT3YNS.TLRV'VSALPIQE-TQD .

‘ WMSGKEZFKCK‘V‘NNKDLEAPTERTISKPKGSVRAEQV’Y‘1’}...‘PPPEEEIVTTKKQV ‘ TLTC.1V1V’T‘DEMPEDTYV’EWTNNGKTLLNYKN'T'ET‘VI‘T.QSTTGSYEVTYSKLRV" w—‘......./ KCABEEBWLK Q1:‘\‘1IESLsEEV1KP1sE‘SLSLTLTVS1TYSTSSTJE A‘1‘1I'1TVVITRQT‘T—‘(xKLsTE‘v‘v‘V’TGYT TTYSGSi‘NYSESTQ11K1S1TRQSSTNQT‘ELQLNSV"TA.SDTA1‘1‘‘1‘1.‘ARGYY111‘1":A 11’EV’YWGQC1'T‘LV’TVSAAS1K11ESVET—‘LAT—‘SSKSTSGGT‘AAT(1C1VKT‘J‘Y'E1’1"?3 V1V’SVSNSGALTSGV’11TEPAVEQSSGTYS1SSVYTVESSSLGTQ"1Y1CNV’N11K E‘SN1KV11KKV’TTEKSCDTLTTTTL1’1‘C1‘1AT‘E1.1(ICESVELT‘EPKPKL‘ITLMTSR11‘1: VLC‘1’V’ V’DVSHEDETIV’KENW1VDG‘VE‘V’T’iNAK.1‘KEREEQYNSTYRV’V’SVLT ‘11..TTQDV‘VLNCKEYKCKV’SNKATPAE‘TEKTTSKAKC:QPKEE’QVYTE1’1‘SK1‘1T.T: KNQVSTTC1VKQT‘YT‘SDTAV’EV‘V’STSGQPE\\‘1KTT-EVLDSDUSTELYSKLT‘ . .."vwn..1....................____.....C-. .I.I.I.I.I.I.I."\A"".V._._.........MM\................................................... ..__........_.....M___.. ..."vv."v"WVWA..._______..........AAAAAAAC.AAAw—.MM"....__........ 11*TYSGQEN1’NEST1xSRV’STTRDTSKNQET‘LQLN‘3'1T115111ATYYCAKGYYCT‘ 11AWTA‘YWGQGT‘L‘VT‘V".SAAK.1‘TET’SV'YE’TA131:‘SAAQTNSMV1‘LGCLV’KGYE‘ "3.3vaPRUKPSER:CNVIA ............................................ GC‘KPCKTVPEVSSA/TITT’T’KPKDVTTTTTTPKVTCV T . SSTKV’DKKTVT’KDC SELT‘TMTTQTNV 1 VVDTSKT)T)PEVQTSVVTVIDDV’EVHTAQTQPRTTQTNSTTRSV LNGKETIKCRVNSAAPPAPTEKTTSKTKGKT’KAPQV‘1TTPPPKEQMAKDKVST TCMTTDPPPEDTTVEVVQVVNGQPAENYKNTQPTMNTNGSV’EVYSKLNV’QKSN 7 AGNTETCSVI’LPTTC1TTTNHHTPKSLSTTSPC1K(SEQTD AAQ.1} ‘ EV’QLQESGPGTVKPSQSLSLTCTV’SGYSTTSC717EA‘sV’HVVTRQEPGKT:LEWMGYT TTYSGGTNYNPSLKSRV STTREPTSKNQTELQLNSVTTAAT}T7.ATYYCARGYYGE C1AVVPPAYWGQGTLVPVSAASTKGPSVPPLAPSSKST7SC1C73TAAT.GETVKDV’EP TEPV’.7VS‘WNSC5IALTSGVHTEPAVT.QSSGT‘17ST.SSVVTVPSSSLGTQT‘17T7CNV7N E i TTKPSNT7KVTEKKVEPKSCDKTHTC7.T’PCPAPET.T.Cs(sPSVPLEPPKPKDTTMTSKT T IPEVTCVV7V7D‘17SHEDPEVKENWYV’T)C1VEV7HNAKTKPREEQYNS"1YRVV7SV7LTTVLHQDVVLNGKEVKCKVSNKALPAPTLKTTSKAKGQPRTPQVYTLPPSRDETTKNQVSLPCLVKG‘TVPSTHAVEVVESNGQPENNYKT7T7PPVLT‘ASDGSETLYSKLE VV'SPST.KSRTSTTRDTSKNQTTLQLNSV7TTEDTA7TV7‘1'C:ARGYYGYG AWEA717VV’1:QGTLVTV SAAKTTAPSVYPLIAPVCGT)TT7GSSVTLGCLV’KGYEP i EPVTLTVVNSGSLSSGVT-TTEPAVTQSDLYTLSSSVTVTSSTWPSQSTTCNVATTPE E ASSTKVDKKTEPKGPTTKPCPPCKCPAPNLLGGT’SVPTEPPKTKTDVT.MTSLSPTV E TCVVVDVSE7DDPE3VQTSVV’EVNN7VEVPTTAQTQTTTREDYNSTTRVV78.AT.PTQTT? QT)VVMSGKLTKCKVNNKT)LPAFTERTTSKPKGSVRAPQVYVLPPPEEEMT7KK QVTLTC‘.MVTTB‘TMPEDTVVEWTNNGK7T7PT \YKNTEPVTDSDGSYEMYSKT.R VEKKNVVVERNSYSCASVATTEGLH\TTTT DNG 105V huCL‘B'f—S? QVQL 77 1. IKGLPVVVTGYT LYSGST7VYSPSTKSRTSTTRDTSKNQPTLGTNSV’TAADTATYYCAKGYYGVG AWEIAYVVGQGTL.V7T7‘17SAASTKGPSVPLAPSSKSTSGGTAAT..GCLV’KDYFPET PVTVSVVNSGALTSGVTTTET)AV’TQSSGT.YSLSSVVTVPSSST.G77TQTYTCNVNTT 7KV77DKKVEPKSC‘DKTTTTCT‘PCPAPET..T.CsGPSVELEPPKPKL)TLMTSR7TP; EV'7TCV’VV'T)‘17SHCDPT’VKTNVVYVDGVEVH‘TAKTKPRECQYNSTYRVV’SVLT; VLTTQDVV’LNGKEYKCKVSNKIALPAPTEKTTSKAKCTQPREPQVYTLPPSRJELT771 SKLTT KNQVSLTCLE7[KGEYPSDTAVTVVTSNGQPENNYKTTPPVLDST‘GSEETY T 252-3 7777PVQVVESGGDLVKPGC KTSCAASGTTTSSYGMSVVVKQTPDKKTEVV777VIACT 7 SSGGSV’TVSPDSVKGRETTSRT)NAKKTL‘1LQMSSLKSET)TATVTY‘7CARTTSV 7‘71 DTSVT)YVVGQGTSVTVSSAKTTAT‘SV'V’PLAPVCGDTTGSSV'TLGCLV’KGYPP EPVTL'T'VVNSGSLSSGVT—TT‘EPAV’LQSDLVCTLSSSVTVTSSTWPSQSTTCNVAHP ASSTKVDKKTEPRGPTTKPCPPCKCTPAPNLLGGPSVETEPPKTKDVLMTSLST‘TV7 TCVVV’DVSEEDPDVQTS‘1‘17EV7NN7V’EVT-T'TAQTQTHREDYNSTLKVVSALT’TQTT QDVVTVTSGKEPKCKVNNKTJLPAPTERTTSKPKGSVKAPQV’YVLPPPEEEMTKK QV’TLTCMVTDPMPEDTYVE.VVTNNGKTELNYKNTET’VLDSDGSV’EMYSKLK 11111111111111.1111111111111111111111111111111.1110111 1111791 Table 6: Full-length light chain amino acid sequences muCD37—3 DTQV’T’TQSPASLSV’SVGETV’ . ., 711177111 NLADGVPSRPS1zSGSGTQYSLKEN11.11311111G1111':QHYw1.1111111(1111111. 1.ETKRADAAP11STTIPPSSEQLTST111113111C1LNNPV’T’KDTNVXWKTDGSTRQ 111111;TDSKDSTYSVTSS11.11.TKDPYERTNSV10311111111STSPTVKS. 1 FNRNEC‘ATSPQ 111 NQ 1011 chCD373 QSPASL§7 " -36— 'NLADGVPSRFSGSGSGTQYSLKFNSLQSEDFGTYYCQHYWGTTWTFGGGTK LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQE SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKE SFNRGEC (SEQ ID NOJ91 ; huCD373 DIQMTQSPSSLSVSVGERVTITCRASENIRSNLAWYQQKPGKSPKLLVNVAT E110 and 1.1) NLADGVPSRESGSGSGTDYSLK1NSLQPEDFGTYYCQHYWGTTWTEGQGTK; 1 LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ; E SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKE SFNRGEC(SEQID NO 1071 = DIV, QSPASLAVSLGQRATISCRASQSVSTSSYSYLY *QQKPGQPPKLLIK 71,,,..,,,,,,,,,,,,,,,,,,,,, EYASNLASGVPARFSGSGSGTDFTLNIHPVEEEDTATYYCQHS GGG TKLEIKRADAAPTVS11PPSSEQLTSGGASVVCFLNNEYPKDINVKWKIDGSE EVKSFNRNEC1‘SEQ1DRQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPINO11181 'Efiéfii‘i‘i‘z'" DIVLTQSPASLAVSLGQRATISCRASQSVSTSSYSYLYWFQQKPGQPPKLLIK YASNLASGVPARFSGSGSGTDETLNILD’VEEEDTATYYCQHSWEIPYTFGGG ETKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA ELQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC1SEQID NO11191 _1 _____ Eiiiii‘ébfil‘fi" QIVLTQSPAIMSASPGEKVTMTCSASSSVTYMHWYQQKSGTSPKRWIYDTS EKLASGVPARFSGGGSGTSYSLTISSMEAEDAATYYCQQWISNPPTFGGGTKL EEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQN GEVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSF E ENRNEC 1SEQ ID NO 110) chCD‘é‘YEEWmW1QIVLTQSPAIMSASPGEKVTMTCSASSSVTYMHWYQQKSGTSPKRWIYDTS_ KLASGVPARFSGGGSG1SYSLTISSMEAEDAATYYCQQWISNPPTFGGGTKLE EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYEREAKVQWKVDNALQSE GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS; 111111 C(SEQ 1DN0111) 38 QSPASMSASPGERVTMTCSASSSVTYMHWYQQKPGTSPKRWXDTS ; EKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWISNPPTFGGGTKLI EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSE GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSE FNRGEC (SEQ ID NO 1 12) i‘ii‘Sfiwm"E"Q1VLTQSPAIMSASPGEKVTMTCSATSSVTYMHWYQQKSGTSPKRWIYDTS KLPYGVPGRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSDNPPTFGSGTKL E EIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSEE?QN TDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSF _E NRNEC (_SEQID NO:113_1_ 1111:5757)‘ EIVLTQSPATMSASPGERVTMTCSATSSVTYMHWYQQKPGQSPKKWHDTS E NLPYGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSDNPPTFGQGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSE . GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSE FNRGEC1SEQID NO 1141 777777 muCD3751 EQIVLTQSPAIMSASPGEKVTMTCSATSSVTYMHWYQQKSGTSPKRWIYDTS ; 1 KLASGVPARFSGSGSGTSYSLTISNMEAEDAATYYCQQWSSNPPTFGSGTKL 1 EIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQN E : GVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSF huCD37-5 1 1EIVLTQSPATMSASPGERVTMTCSATSSVTYMHWYQQKPGQSPKRWIYDTS KLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGQGTKL EIKRTVAAPSVFIFPPSDEQLEKSGTASVVCLLNNFYPREAKVQWKVDNALQSWE _37_ EGNSQE‘WTEQBS‘Kfifififs'sTLTLsflfiYEWKfiKV§Kf§i§fifi65§§fifii§W4 _SEQ ID NO: 116.: 1 muCD37-56 QIVLTQSPAFMSASPGDKVTMTCSASSSVTYMHWYQQKSGTSPKRWIYDTS E PARFSGGGSGTSYSLTISTMEAEDAATYYCQQWISDPP‘EtGGGTKL EIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQN WTDQDSKBSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSF NRNEC (SEQ ID NO 117) Ehucfig‘fig"""""""""‘E"DIVLTQSPAFMSASPGEKVTMTCSASSSVTYMHWYQQKPDQSPKRWIYDTS NLASGVPSRFSGGGSGTDYSLTISSMEAEDAATYYCQQWISDPPTFGQGTKL; EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSE: GENSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKS ‘FNRGEC @139 ID NO 1 18) muCD3757 QlVLTQSPAIMSASPGEKVTMTCSATSSVTYMHWYQQKSGTSPKRWIYDTS E KLASGVPARFSGSGSGTSYSLTISSMLAEDAATYYCQQWSDNPPTFGSGTKL EIEKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQN GVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSF huCD3757 EIVLTQSPATMSASPGERVWSSVTYMHWYQQKPGQSPRRWIYDTS NLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSDNPPTFGQGTKL; EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNPYPREAKVQWKVDNALQSE GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSE FNRGEC(SEQ ID NO:___1_2__(_)) EEEEEEEEEEEEEEEEE 252-3 EDIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTS EKLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNALPWTFGGGTKL EELKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQ : NGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKS FNRNEC"(SEQIDNQ: 180) Also provided are polypeptides that comprise: (a) a ptide having at least about 90% at least about 90% sequence identity to SEQ ID NOs:88-104 or 179; and/or (b) a polypeptide having sequence identity to SEQ ID NOS:105-120 or 180. In certain embodiments, the polypeptide comprises a polypeptide having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% ce ty to SEQ ID NOs:88-l20, 179, or 180. Thus, in certain embodiments, the polypeptide comprises (a) a polypeptide having at least about 95% sequence identity to SEQ ID NOs:88- 104 or 179, and/or (b) a polypeptide having at least about 95% sequence identity to SEQ ID NOs:105-120 or 180. In certain embodiments, the polypeptide comprises (a) a polypeptide having the amino acid sequence of SEQ ID NOs:88-IO4 or 179; and/or (b) a ptide having the amino acid sequence of SEQ ID NOs:105-120 or 180. In certain embodiments, the polypeptide is an antibody and/or the polypeptide specifically binds CD37. In certain embodiments, the ptide is a murine, chimeric, or humanized arétibody that specifically binds CD37. In certain embodiments, the polypeptide having a certain percentage of ce identity to SEQ ID NOs:88-120, 179, or 180 differs from SEQ ID NOs:88- 120, 179, or 180 by conservative amino acid substitutions only.

] In certain embodiments, the CD37 antibody can be the antibody produced from a hybridoma selected from the group consisting of consisting of ATCC Deposit Designation PTA—10664, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10665, deposited with the ATCC -38— on February 18, 2010, ATCC Deposit Deisgnation 666, deposited with the ATCC on February 18, ‘10, ATCC Deposit Designation PTA~10667 deposited with the ATCC on February 18, 2010, ATCC Deposit Designation traumas, deposited with the ATCC on February 18, 20W, ATCC Deposit Designation i0669, deposited with the ATCC on February 18, 2010, and ATCC Deposit Designation PTA»10670, deposited with the ATCC. on February 18, 2010 {American Type Culture Collection (ATCC) at 10801 University Boulevard, Manassas, Virginia 20E iii). In certain embodiments, the antibody comprises the VH—CDRS and the VI..~CDRS of the dy produced from a bydridoma selected from the group consisting 10665, P13840666, PTA-10667, 140668, PTA~10669, and PTA-10670.

In certain embodiments, the CD37 dy can comprise a light chain encoded by the inant plasmid DNA phuCD37-3LC (ATCC Deposit Designation PTA-10722, deposited with the ATCC on March 18, 2010). In n embodiments, the CD37 antibody can comprise a heavy chain encoded by the inant plasmid DNA phuCD37—3HCV.1.0 (ATCC Deposit Designation 723, deposited with the ATCC on March 18, 2010). In certain embodiments, the CD37 antibody can comprise a light chain encoded by the recombinant plasmid DNA 7-3LC (PTA-10722) and a heavy chain encoded by the recombinant plasmid DNA phuCD37-3HCV.l.0 (PTA—10723). In certain embodiments, the CD37 antibody can se the VL—CDRs encoded by the recombinant plasmid DNA phuCD37-3LC (PTA-10722) and the VH-CDRs encoded by the inant plasmid DNA phuCD37-3HCV.1.0 (PTA- 10723).

Monoclonal dies can be prepared using hybridoma s, such as those described by Kohler and Milstein (1975) Nature 256:495. Using the hybridoma method, a mouse, hamster, or other appropriate host animal, is immunized as described above to elicit the production by lymphocytes of antibodies that will cally bind to an immunizing antigen. Lymphocytes can also be immunized in vitro. Following immunization, the lymphocytes are isolated and fused with a suitable myeloma cell line using, for example, polyethylene glycol, to form hybridoma cells that can then be selected away from unfused lymphocytes and myeloma cells. Hybridomas that produce monoclonal antibodies directed specifically against a chosen antigen as determined by immunoprecipitation, immunoblotting, or by an in vitro binding assay (e.g. mmunoassay (RIA); enzyme-linked immunosorbent assay (ELISA)) can then be propagated either in vitro culture using standard methods (Goding, Monoclonal Antibodies: Principles and Practice, Academic Press, 1986) or in vivo as ascites tumors in an animal. The monoclonal antibodies can then be purified from the culture medium or ascites fluid as described for polyclonal dies above.

Alternatively monoclonal antibodies can also be made using recombinant DNA methods as described in US. Patent 4,816,567. The polynucleotides encoding a monoclonal antibody are isolated from mature B—cells or hybridoma cell, such as by RT-PCR using oligonucleotide primers that specifically ~39— amplify the genes encoding the heavy and light chains of the dy, and their sequence is determined using conventional procedures. The isolated polynucleotides encoding the heavy and light chains are then cloned into suitable expression vectors, which when ected into host cells such as E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce globulin protein, monoclonal antibodies are generated by the host cells. Also, recombinant monoclonal antibodies or fragments f of the desired species can be ed from phage y libraries expressing CDRs of the desired species as described (McCafferty et al., 1990, Nature, 3482552- 554; Clackson et al., 1991, Nature, 352:624-628; and Marks et al., 1991, J. Mol. Biol., 222:581-597).

The polynucleotide(s) encoding a monoclonal antibody can further be modified in a number of different manners using recombinant DNA technology to generate alternative antibodies. In some embodiments, the constant domains of the light and heavy chains of, for example, a mouse monoclonal antibody can be substituted 1) for those regions of, for example, a human antibody to generate a chimeric antibody or 2) for a non-immunoglobulin polypeptide to generate a fusion antibody. In some ments, the constant regions are truncated or removed to generate the desired dy fragment of a monoclonal antibody. Site-directed or high-density mutagenesis of the variable region can be used to optimize specificity, affinity, etc. of a monoclonal antibody.

In some embodiments, the monoclonal antibody against the human CD37 is a humanized antibody. In certain ments, such antibodies are used therapeutically to reduce antigenicity and HAMA (human ouse antibody) responses when administered to a human subject. Humanized antibodies can be produced using various techniques known in the art. In certain alternative embodiments, the antibody to CD37is a human antibody.

Human antibodies can be directly prepared using s techniques known in the art.

Immortalized human B lymphocytes immunized in vitro or isolated from an zed individual that produce an antibody directed against a target antigen can be ted (See, e.g., Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boemer et al., 1991, J. Immunol, 147 (1)186— 95; and US. Patent 5,750,373). Also, the human antibody can be selected from a phage library, where that phage library expresses human antibodies, as described, for e, in Vaughan et al., 1996, Nat.

Biotech., 14:309-314, Sheets et al., 1998, Proc. Nat’l. Acad. Sci., 95:6157162, Hoogenboom and Winter, 1991, J. Mol, Biol., 1, and Marks et al., 1991, J. Mol. Biol, 222:581). Techniques for the generation and use of antibody phage libraries are also bed in US. Patent Nos. 5,969,108, 6,172,197, 5,885,793, 6,521,404; 6,544,731; 6,555,313; 6,582,915; 6,593,081; 6,300,064; 068; 6,706,484; and 7,264,963; and Rothe et al., 2007, J. Mol. Bio., 376:1182 (each of which is incorporated by nce in its entirety). Affinity maturation strategies and chain shuffling strategies (Marks et al., 1:992, Bio/Technolngy 10:779»783, orated by reference in its entirety) are known in the art and can be employed to generate high att‘inity human antibodies.

] Humanized dies can also be made in transgenic mice containing human immunoglobulin loci that are capable upon immunization of producing the full repertoire of human antibodies in the absence of endogenous immunoglobulin tion. This approach is described in US.

Patents 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016.

This invention also encompasses bispecific antibodies that specifically recognize a CD37.

Bispecific antibodies are antibodies that are e of specifically recognizing and binding at least two different es. The different epitopes can either be within the same molecule (e.g. the same CD3 7) or and bind a on different les such that both, for e, the antibodies can specifically recognize CD37 as well as, for example, 1) an effector molecule on a leukocyte such as a T—cell receptor (e.g. CD3) below. or PC receptor (e.g. CD64, CD32, or CD16) or 2) a cytotoxic agent as described in detail Exemplary bispecific dies can bind to two different epitopes, at least one of which originates in a polypeptide of the invention. Alternatively, an anti-antigenic arm of an immunoglobulin molecule can be combined with an arm which binds to a triggering molecule on a leukocyte such as a T cell receptor molecule (e.g. CD2, CD3, CD28, or B7), or Fc receptors for IgG so as to focus cellular defense mechanisms to the cell expressing the particular n. Bispecific antibodies can also be used to direct cytotoxic agents to cells which express a particular antigen. These antibodies possess an antigen- binding arm and an arm which binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPTA, DOTA, or TETA. ques for making bispecific antibodies are common in the art (Millstein et al., 1983, Nature 7—539; Brennan et al., 1985, Science 229:81; Suresh et al, 1986, Methods in Enzymol. 121:120; cker et al., 1991, EMBO J. 10:3655—3659; Shalaby et al., 1992, J. Exp. Med. 175:217-225; Kostelny et al., 1992, J. Immunol. 148:1547-1553; Gruber et al., 1994, J. Immunol. 68; and US. Patent 5,731,168). Antibodies with more than two valencies are also contemplated.

For example, trispecific antibodies can be prepared (Tutt et al., J. Immunol. 147:60 ). Thus, in certain embodiments the antibodies to CD37 are multispecific.

In certain embodiments are provided an antibody fragment to, for example, increase tissue penetration. Various techniques are known for the production of antibody fragments. Traditionally, these fragments are derived via lytic digestion of intact antibodies (for example Morimoto et al., 1993, Journal of Biochemical and Biophysical Methods 242107-117; Brennan et al., 1985, Science, 229281). In n embodiments, antibody fragments are produced recombinantly. Fab, Fv, and scFv antibody fragments can all be expressed in and secreted from E. coli or other host cells, thus allowing the production of large amounts of these fragments. Such antibody fragments can also be isolated from the antibody phage libraries sed above. The antibody fragment can also be linear antibodies as described in US. Patent 5,641,870, for example, and can be monospecific or bispecific. Other techniques for the production of antibody fragments will be nt to the d practitioner.

According to the t invention, techniques can be adapted for the productior’: of single- chain antibodies specific to CD37 (see US. Pat. No. 4,946,778). In addition, methods can be adapted for the construction of Fab expression libraries (Huse, et al., Science 246:1275-1281 (1989)) to allow rapid and effective fication of monoclonal Fab fragments with the desired specificity for CD37, or derivatives, fragments, analogs or homologs thereof. Antibody fragments can be ed by ques in the art including, but not limited to: (a) a F(ab')2 fragment produced by pepsin digestion of an antibody molecule; (b) a Fab fragment ted by reducing the disulfide bridges of an F(ab')2 fragment, (c) a Fab fragment generated by the treatment of the antibody molecule with papain and a reducing agent, and (d) Fv fragments.

It can further be desirable, especially in the case of antibody fragments, to modify an antibody in order to increase its serum half-life. This can be achieved, for example, by incorporation of a salvage receptor binding epitope into the antibody fragment by mutation of the appropriate region in the antibody fragment or by incorporating the epitope into a peptide tag that is then fused to the dy fragment at either end or in the middle (e.g., by DNA or peptide synthesis). conjugate antibodies are also within the scope of the present invention.

Heteroconjugate antibodies are composed of two covalently joined antibodies. Such dies have, for example, been ed to target immune cells to unwanted cells (US Pat. No. 4,676,980). It is plated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking . For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methylmercaptobutyrimidate.

For the purposes of the present invention, it should be appreciated that modified antibodies can comprise any type of variable region that provides for the association of the antibody with the polypeptides of a human CD37. In this regard, the variable region can comprise or be derived from any type of mammal that can be induced to mount a humoral response and generate immunoglobulins against the desired n. As such, the variable region of the modified antibodies can be, for example, of human, murine, man primate (e.g. cynomolgus monkeys, macaques, etc.) or lupine origin. In some embodiments both the variable and constant regions of the modified globulins are human. In other embodiments the variable regions of compatible antibodies (usually derived from a non-human source) the can be engineered or specifically tailored to improve the binding properties or reduce genicity of the molecule. In this respect, variable regions useful in the present invention can be humanized or otherwise altered through the inclusion of imported amino acid sequences. {00142} In n embodiments, the le domains in both the heavy and light chains are d by at least partial replacement of one or more CDRs and, if necessary, by partial framework region replacement and ce changing. gh the CDRs can be derived from an antibody of the same class or even subclass as the antibody from which the framework regions are derived, it is ged that the CDRs will be derived from an dy of different class and possibly from an antibody from a different s. It is not alway necessary to replace all of the CDRs with the complete CDRs from the donor variable region to transfer the antigen binding capacity of one variable domain to another. Rather, in some cases it is only necessary to transfer those residues that are necessary to maintain the activity of the antigen binding site. Given the explanations set forth in U.S. Pat. Nos. 5,585,089, 5,693,761 and ,693,762, it will be well within the competence of those skilled in the art, either by carrying out routine experimentation or by trial and error testing to obtain a functional antibody with reduced immunogenicity.

Alterations to the variable region hstanding, those skilled in the art will appreciate that the d antibodies of this invention will comprise antibodies (e.g., full-length antibodies or immunoreactive fragments thereof) in which at least a fraction of one or more of the constant region domains has been deleted or otherwise altered so as to provide desired biochemical characteristics such as reduced serum ife when compared with an dy of approximately the same immunogenicity comprising a native or unaltered constant region. In some embodiments, the constant region of the d dies will comprise a human constant . ations to the constant region compatible witl‘: this invention comprise ons, deletions or substitutions of one or more amino acids in one or more domains. That is, the modified antibodies disclosed herein can comprise tions or modifications to one or more of the three heavy chain constant domains (CH1, CH2 or CH3) and/or to the light chain constant domain (CL). In some embodiments, modified constant regions wherein one or more domains are partially or entirely deleted are contemplated. In some embodiments, the modified dies will comprise domain deleted constructs or variants wherein the entire CH2 domain has been removed (ACH2 constructs). In some embodiments, the omitted constant region domain will be replaced by a short amino acid spacer (e.g. 10 residues) that provides some of the molecular flexibility typically imparted by the absent constant region.

] Besides their configuration, it is known in the art that the constant region mediates several effector ons. For example, binding of the C1 component of complement to antibodies activates the complement system. Activation of ment is important in the opsonisation and lysis of cell pathogens. The activation of complement also stimulates the inflammatory response and can also be involved in autoimmune hypersensitivity. Further, antibodies bind to cells via the Fc region, with a PC receptor site on the antibody Fc regior: binding to a Fc receptor (FcR) on a cell. There are a number of Fc receptors which are specific for different classes of antibody, ing IgG (gamma receptors), IgE (eta receptors), IgA (alpha ors) and IgM (mu ors). Binding of antibody to Fc receptors on cell surfaces triggers a number of important and diverse biological responses including engulfment and destruction of antibody-coated particles, clearance of immune complexes, lysis of antibody-coated target ~41\l—J eeliis by kiiier cells (called antibody-dependent cell—mediated cytotoxicity, or ADCC), reiease of inflammatory mediators, placental transfer and control ofimrnunogiobuiin production.

{GM-’35} in certain embodiments, the CD37~binding antibodies provide for altered effector functions that, in turn, affect the. biologicai profiie of the. administered dy. For example, the deietion or inactivation (through point mutations or other means) of a constant region domain can reduce Fe receptor binding of the circulating modified antibody. in other cases, it. can be that constant region modifications, consistent with this invention, moderate compiement binding and thus reduce the serum half iife and nonspecific association of a conjugated cytotoxin. Yet other cations of the constant region can be used to eliminate disulfide linkages or oligosaccharide moieties that allow for enhanced localization due to increased antigen specificity or dy flexibility. Similarly, modifications to the constant region in accordance with this invention can easily be made using well known biochemical or molecular engineering techniques well within the purview of the d artisan. gotiiztti} In certain embodiments, a CD3 7—binding agent that is an antibody does not have one or more effector functions. For instance, in some embodiments, the antibody has no antibody—dependent cellular cytotoxicity (ADCC) activity and/or no complement-dependent cytotoxicity (CDC) ty. In certain embodiments, the antibody does not bind to an F0 receptor and/or complement factors. In certain embodiments, the antibody has no effector on.

It will be noted that in certain embodiments, the modified antibodies can be ered to fuse the CH3 domain directly to the hinge region of the respective modified antibodies. In other ucts it can be desirable to e a peptide spacer between the hinge region and the modified CH2 and/or CH3 domains. For example, compatible constructs could be sed wleerein the CH2 domain has been deleted and the remaining CH3 domain (modified or fied) is joined to the hinge region with a 5—20 amino acid spacer. Such a spacer can be added, for instance, to ensure that the regulatory elements of the constant domain remain free and ible or that the hinge region remains flexible.

However, it should be noted that amino acid spacers can, in some cases, prove to be immunogenic and elicit an unwanted immune response against the construct. Accordingly, in certain embodiments, any omitted ther, so as to spacer added to the construct will be relatively non-immunogenic, or even maintain the d biochemical qualities of the modified antibodies.

Besides the on of whole constant region domains, it will be appreciated that the antibodies of the present invention can be provided by the l deletion or substitution of a few or even a single amino acid. For example, the mutation of a single amino acid in selected areas of the CH2 domain can be enough to substantially reduce Fc binding. Similarly, it can be ble to simply delete that part of one or more constant region domains that control the effector function (e.g. complement CLQ binding) to be modulated. Such partial deletions of the nt regions can improve selected characteristics of the antibody (serum ife) while leaving other desirable functions associated with the subject constant region domain intact. Moreover, as alluded to above, the constant regions of the disclosed antibodies can be modified through the mutation or substitution of one or more amino acids that es the profile of the resulting construct. In this respect it can be le to disrupt the activity provided by a ved binding site (e.g. Fc binding) while substantially maintaining the configuration and immunogenic profile of the modified antibody. Certain embodiments can comprise the addition of characteristics such as sing or one or more amino acids to the nt region to enhance desirable increasing effector on or provide for more cytotoxin or carbohydrate attachment. In such embodiments it can be desirable to insert or replicate specific sequences derived from selected constant region s.

The present invention further embraces variants and equivalents which are substantially homologous to the chimeric, humanized and human antibodies, or antibody fragments thereof, set forth herein. These can contain, for example, conservative substitution mutations, i.e. the substitution of one or more amino acids by similar amino acids. For example, conservative substitution refers to the substitution acid of an amino acid with another within the same general class such as, for e, one acidic amino with another acidic amino acid, one basic amino acid with another basic amino acid or one neutral amino acid by another neutral amino acid. What is intended by a conservative amino acid substitution is well known in the art.

The polypeptides of the present invention can be recombinant polypeptides, natural polypeptédes, or synthetic polypeptides comprising an antibody, or fragment thereof, against a human CD37. It will be recognized in the art that some amino acid sequences of the invention car; be varied without significant effect of the structure or function of the protein. Thus, the invention further includes variations of the ptides which show substantial activity or which include regions of an antibody, or fragment thereof, against CD37 protein. Such mutants include deletions, insertions, inversions, repeats, and type substitutions.

The ptides and analogs can be further modified to contain additional chemical moieties the biological half not normally part of the protein. Those tized moieties can improve the solubility, life or absorption of the protein. The moieties can also reduce or eliminate any desirable side effects of proteins and the like. An overview for those moieties can be found in REMINGTON'S PHARMACEUTICAL SCIENCES, 20th ed., Mack Publishing Co., Easton, PA (2000).

] The isolated polypeptides bed herein can be produced by any suitable method known in the art. Such s range from direct protein tic methods to constructing a DNA sequence ng isolated polypeptide sequences and expressing those sequences in a le transformed host.

In some embodiments, a DNA sequence is constructed using recombinant technology by isolating or synthesizing a DNA sequence encoding a wild—type n of interest. Optionally, the sequence can be mutagenized by site-specific mutagenesis to provide onal analogs thereof. See, e.g. Zoeller et al., Proc. Nat’l. Acad. Sci. USA 81:5662-5066 (1984) and US. Pat. No. 4,588,585.

In some embodiments a DNA ce encoding a polypeptide of interest would be constructed by chemical synthesis using an oligonucleotide synthesizer. Such oligonucleotides can be designed based on the amino acid sequence of the desired polypeptide and selecting those codons that are favored in the host cell in which the recombinant polypeptide of interest will be produced. rd methods can be applied to synthesize an isolated polynucleotide sequence encoding an isolated polypeptide of interest. For example, a complete amino acid sequence can be used to construct a back— translated gene. Further, a DNA oligomer containing a nucleotide sequence coding for the particular isolated polypeptide can be synthesized. For example, several small ucleotides coding for portions of the desired polypeptide can be synthesized and then ligated. The individual ucleotides typically contain 5' or 3' overhangs for complementary asembly.

Once assembled (by synthesis, irected mutagenesis or another method), the polynucleotide sequences ng a particular ed ptide of interest will be inserted into an expression vector and operatively linked to an expression control ce appropriate for expression of the protein in a desired host. Proper assembly can be confirmed by nucleotide sequencing, restriction mapping, and expression of a ically active polypeptide in a suitable host. As is well known in the art, in order to obtain high expression levels of a transfected gene in a host, the gene must be operatively linked to transcriptional and ational expression control sequences that are functional in the chosen expression host.

In certain ments, recombinant expression vectors are used to amplify and express DNA encoding antibodies, or fragments thereof, against human CD37. Recombinant expression vectors are replicable DNA constructs which have synthetic or cDNA-derived DNA nts encoding a polypeptide chain of an anti-CD37 antibody, or fragment thereof, operatively linked to suitable transcriptional or translational regulatory elements derived from mammalian, microbial, viral or insect elements having genes. A transcriptional unit lly comprises an assembly of (1) a genetic element or a regulatory role in gene expression, for example, transcriptional ers or enhancers, (2) a structural or coding sequence which is transcribed into mRNA and translated into n, and (3) appropriate transcription and translation initiation and termination sequences, as described in detail below. Such regulatory elements can include an operator sequence to control transcription. The y to ate in a host, usually conferred by an origin of replication, and a selection gene to facilitate recognition of transformants can additionally be incorporated. DNA regions are operatively linked when they are functionally related to each other. For example, DNA for a signal peptide (secretory leader) is operatively linked to DNA for a polypeptide if it is expressed as a precursor which participates in the secretion of the polypeptide; a promoter is operatively linked to a coding sequence if it controls the transcription of the ._ 45 a if it is oned so as to sequence; or a ribosome binding site is operatively linked to a coding ce permit ation. Structural elements intended for use in yeast expression systems include a leader sequence enabling extracellular secretion of translated protein by a host cell. Alternatively, where recombinant protein is expressed without a leader or transport sequence, it can e an N-terminal methionine residue. This residue can ally be subsequently d from the sed recombinant protein to provide a final product.

The choice of expression control sequence and expression vector will depend upon the choice of host. A wide variety of expression host/vector combinations can be employed. Useful expression vectors for otic hosts, include, for example, vectors comprising expression control sequences from SV40, bovine papilloma virus, adenovirus and cytomegalovirus. Useful expression s fer bacterial hosts include known bacterial plasmids, such as plasmids from Esherichia coli, including pCR l, pBR322, pMB9 and their derivatives, wider host range plasmids, such as M13 and tous single-stranded DNA phages.

Suitable host cells for expression of a CD37-binding polypeptide or antibody (or a CD37 protein to use as an antigen) include prokaryotes, yeast, insect or higher eukaryotic cells under the control of appropriate promoters. Prokaryotes include gram ve or gram ve organisms, for example E. coli or bacilli. Higher eukaryotic cells include established cell lines of mammalian origin as described below. Cell-free translation systems could also be employed. Appropriate cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cellular hosts are described by Pouwels et al.

(Cloning Vectors: A Laboratory Manual, Elsevier, N.Y., 1985), the relevant disclosure of which is hereby orated by reference. Additional information regarding methods of protein production, including antibody production, can be found, e.g., in US. Patent Publication No. 2008/0187954, US. Patent Nos. 6,413,746 and 6,660,501, and International Patent Publication No. WO 04009823, each of which is hereby incorporated by reference herein in its entirety.

Various mammalian or insect cell culture systems are also advantageously employed to express recombinant protein. Expression of recombinant proteins in mammalian cells can be performed because such proteins are generally correctly folded, appropriately modified and completely functional.

Examples of suitable mammalian host cell lines include the COS-7 lines of monkey kidney cells, described by Gluzman (Cell , 1981), and other cell lines capable of expressing an appropriate vector including, for e, L cells, C127, 3T3, Chinese hamster ovary (CHO), HeLa and BHK cell lines. Mammalian sion vectors can comprise nscribed elements such as an origin of replication, a le promoter and enhancer linked to the gene to be expressed, and other 5' or 3' flanking nontranscribed sequences, and 5' or 3' nontranslated sequences, such as necessary ribosome binding sites, a polyadenylation site, splice donor and acceptor sites, and transcriptional termination sequences.

Baculovirus systems for production of heterologous proteins in insect cells are reviewed by Luckow and Summers, Bio/Technology 6:47 (1988).

The proteins produced by a transformed host can be purified according to any suitable method. Such rd methods include chromatography (e.g., ion exchange, affinity and sizing column tography), centrifugation, differential solubility, or by any other standard technique for n purification. y tags such as hexahistidine, maltose binding domain, influenza coat sequence and glutathione-S-transferase can be attached to the protein to allow easy ation by passage over an appropriate affinity . Isolated proteins can also be physically characterized using such techniques as proteolysis, r magnetic resonance and x-ray crystallography.

For example, supematants from systems which secrete recombinant protein into culture media can be first trated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. Following the concentration step, the concentrate can be applied to a suitable purification matrix. Alternatively, an anion exchange resin can be employed, for e, a matréx or substrate having pendant diethylaminoethyl (DEAE) groups. The matrices can be acrylamide, agarose, dextran, cellulose or other types commonly employed in protein purification.

Alternatively, a cation exchange step can be employed. Suitable cation exchangers include various insoluble matrices comprising sulfopropyl or carboxymethyl groups. y, one or more reversed-phase high performance liquid chromatography (RP—HPLC) steps employing hydrophobic RP-HPLC media, be ed to further purify a CD37- e.g., silica gel having pendant methyl or other aliphatic groups, can binding agent. Some or all of the foregoing purification steps, in various combinations, can also be employed to e a homogeneous recombinant protein.

Recombinant protein produced in bacterial culture can be isolated, for example, by initial extraction from cell pellets, followed by one or more concentration, salting—out, aqueous ion exchange or size exclusion chromatography steps. High performance liquid chromatography (HPLC) can be employed for final purification steps. Microbial cells employed in expression of a recombinant protein can be disrupted by any convenient , including freeze-thaw cycling, sonication, mechanical disruption, or use of cell lysing agents.

] Methods known in the art for purifying antibodies and other proteins also include, for example, those described in US. Patent Publication No. 2008/0312425, 2008/0177048, and 2009/0187005, each of which is hereby incorporated by reference herein in its entirety.

In certain embodiments, the CD37—binding agent is a polypeptide that is not an antibody. A variety of methods for fying and producing non-antibody polypeptides that bind with high affinity to a n target are known in the art. See, e.g., Skerra, Curr. Opin. Biotechnol, 18:295-304 (2007), Hosse et al., Protein Science, 15:14-27 (2006), Gill et al., Curr. Opin. hnol, 17:653-658 (2006), Nygren, FEBS J., 275:2668-76 (2008), and , FEBS J., 275:2677-83 , each ofwhich is incorporated by reference herein in its entirety. In certain embodiments, phage y techeology has been used to identify/produce the CD37-binding polypeptide. In certain embodiments, the polypeptide ses a protein scaffold of a type selected from the group consisting of protein A, a lipocalin, a fibronectin domain, an ankyrin sus repeat domain, and thioredoxin.

In some embodiments, the agent is a non-protein molecule. In certain embodiments, the agent is a small molecule. Combinatorial chemistry libraries and techniques useful in the identification of non- protein CD37-binding agents are known to those d in the art. See, e.g., Kennedy et al., J. Comb.

Chem, 10:345—3 54 (2008), Dolle et al, J. Comb. Chem, 9:855—902 (2007), and Bhattacharyya, Curr. Med.

Chem., 8:1383—404 (2001), each of which is incorporated by reference herein in its entirety. In certain further embodiments, the agent is a carbohydrate, a glycosaminoglycan, a glycoprotein, or a proteoglycan. in certain ments, the agent is a nucleic acid aptamer. rs are polynucleotide molecules that have been selected (e.g., from random or mutagenized pools) on the basis of their ability to bind to another molecule. In some embodiments, the aptamer comprises a DNA cleotide. In certain alternative embodiments, the aptamer comprises an RNA polynucleotide. In certain embodiments, the aptamer comprises one or more modified nucleic acid residues. Methods of ting and screening nucleic acid aptamers for binding to proteins are well known in the art. See, e.g., U.S. Patent No. ,270,163, U.S. Patent No. 5,683,867, U.S. Patent No. 5,763,595, U.S. Patent No. 6,344,321, U.S. Patent No. 7,368,236, U.S. Patent No. 5,582,981, U.S. Patent No. 5,756,291, US. Patent No. 5,840,867, U.S.

Patent No. 7,312,325, U.S. Patent No. 7,329,742, ational Patent Publication No. WO 02/077262, International Patent Publication No. WO 03/070984, U.S. Patent Application Publication No. 2005/0239134, U.S. Patent ation Publication No. 2005/0124565, and U.S. Patent Application Publication No. 2008/022773 5, each of which is incorporated by reference herein in its entirety.

III. Immunoconjugates The present ion is also directed to ates (also referred to herein as immunoconjugates), comprising the anti-CD37 antibodies, antibody fragments, and their functional equivalents as disclosed herein, linked or conjugated to a drug or prodrug. Suitable drugs or prodrugs are known in the art. The drugs or prodrugs can be cytotoxic agents. The cytotoxic agent used in the cytotoxic conjugate of the present ion can be any compound that results in the death of a cell, or induces cell death, or in some manner decreases cell viability, and includes, for example, maytansinoids and maytansinoid s. Other suitable cytotoxic agents are for example benzodiazepines, taxoids, CC- 1065 and CC-1065 analogs, duocarmycins and duocarmycin analogs, enediynes, such as calicheamicins, atin and dolastatin s including auristatins, tomaymycin derivaties, ycin derévaties, methotrexate, cisplatin, carboplatin, daunorubicin, doxombicin, vincristine, vinblastine, lan, mitomycin C, chlorambucil and morpholino doxorubicin.

Such conjugates can be prepared by using a linking group in order to link a drug or prodrug to the antibody or functional equivalent. Suitable linking groups are well known in the art and include, for example, disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups.

The drug or prodrug can, for example, be linked to the anti-CD37 antibody or fragment thereof through a disulfide bond. The linker le or crosslinking agent comprises a reactive chemical thereof. The reactive chemical groups for group that can react with the anti-CD37 antibody or fragment reaction with the cell-binding agent can be N-succinimidyl esters and N—sulfosuccinimidyl esters.

Additionally the linker le comprises a reactive chemical group, which can be a dithiopyridyl group that can react with the drug to form a disulfide bond. Linker molecules include, for example, N- imidyl 3-(2-pyridyldithio) propionate (SPDP) (see, e.g., Carlsson et al., Biochem. J., 173: 723-737 (1978)), N—succinimidyl 4-(2-pyridyldithio)butanoate (SPDB) (see, e.g., US. Patent No. 4,563,304), N- succinimidyl 4-(2—pyridyldithio)2—sulfobutanoate (sulfo—SPDB) (see US Publication 'No. 74713) , N—succinimidyl 4-(2-pyridyldithio) pentanoate (SPP) (see, e.g., CAS Registry number 341498-08—6), 2- iminothiolane, or acetylsuccinic anhydride. For example, the antibody or cell binding agent can be d with crosslinking reagents and the antibody or cell binding agent containing free or protected thiol groups thus derived is then d with a disulfide- or thiol-containing maytansinoid to produce conjugates. The conjugates can be purified by chromatography, including but not limited to HPLC, size- exclusion, adsorption, ion ge and affinity capture, dialysis or tangential flow filtration.

In another aspect of the present invention, the D37 antibody is linked to cytotoxic drugs via disulfide bonds and a polyethylene glycol spacer in enhancing the potency, solubility or the efficacy of the immunoconjugate. Such cleavable hydrophilic linkers are described in W02009/0134976. The onal benefit of this linker design is the desired high r ratio and the minimal aggregation of the dy-drug conjugate. Specifically contemplated in this aspect are conjugates of cell-binding agents and drugs linked via disulfide group (-S-S—) bearing polyethylene glycol s ((CH2CH20)H=H4) with a narrow range of drug load of 2-8 are described that show vely high potent biological activity toward cells and have the desired biochemical properties of high conjugation yield and high r ratio with minimal protein aggregation.

Specifically contemplated in this aspect is an anti-CD37 antibody drug conjugate of formula (I) or a conjugate of formula (1'): CB—[X1+CH24:H2CHHNY—D]m (I) -CH2—CHZO~~~)n—X1]m-CB (1') wherein: CB ents an anti-CD37 antibody or fragment; D represents a drug; X represents an tic, an aromatic or a heterocyclic unit attached to the cell—binding agent via a thioether bond, an amide bond, at carbamate bond, or an ether bond; Y represents an aliphatic, an aromatic or a heterocyclic unit attached to the drug Via a disuifide bond; {39:75} i is =3 or t; {00176} m is an integer from 2 to 8; and {00177} n is an integer from 1 to 24. {00178} In some embodiments, m is an integer from 2 to 6. {00:79] In some embodiments, m is an integer from 3 to 5. {00180} In some embodiments, n is an integer form 2 to 8. Aitematively, as disclosed in, for example, US. Patent No. 6,441,163 and 7,368,565, the drug can be first modified to introduce a reactive ester suitable to react with a cell-binding agent. Reaction of these drugs containing an activated linker moiety with a cell-binding agent provides another method of producing a cell-binding agent drug conjugate. sinoids can also be linked to anti—CD37 dy or nt using PEG linking groups, as set forth for example in US. Patent 6,716,821. These PEG non-cleavable linking groups are soluble both in water and in non-aqueous solvents, and can be used to join one or more cytotoxic agents to a cell binding agent. ary PEG linking groups include heterobifunctional PEG linkers that react with cytotoxic agents and cell binding agents at te ends of the linkers through a functional sulthydryl or disulfide of the synthesis of a cytotoxic group at one end, and an active ester at the other end. As a general example conjugate using a PEG linking group, re "erence is again made to US Patent 6,716,821 which is ineoiporated entirely by reference herein. Synthesis begins with the on of one or more cytotoxic agents hearing a reactive PEG moiety with a ceii—‘oinding agent, resulting in dispiacernent of the terminal active ester of each reactive PEG moiety by an amino acid residue of the ceh binding agent, to yield a cytotoxic conjugate comprising one or more cytotoxic agents eovaientiy handed to a eeii binding agent through 21 PEG iinking group, Alternativciy, the cell binding can be modified with the hifiinctionai PEG crossiinker to introduce a reactive- disultide moiety (such as a pyridyidisttifide), which can "then he treated with a thioi—eontaining nsinoid to e a conjugate. in another method, the ceii binding can he modified with the nctionai PEG crosstinker to introduce a thiol moiety which can then. can he treated with 2t reactive disuttide-containing tnaytansinoid (such as a idisuitide), to provide a conjugate. dy-maytansinoid ates with non—cleavable links can also be prepared. Such crosslinkers are described in the art (see US ation No. 20050169933) and include but are not limited to, N—succinimidyl 4—{maleimidomethyl) cyclohexanecarboxylate (SMCC). In some embodiments, the antibody is modified with crosslinking reagents such as succinimidyl 4-(N-maleimidomethyl)— cyclohexane—l—carboxylate , sulfo-SMCC, maleimidobenzoyl—N-hydroxysuccinimide ester (MBS), sulfo-MBS or imidyl-iodoacetate, as described in the literature, to introduce l-lO reactive Hashida et al, J. Applied Biochem., 56—63 groups (Yoshitake et al, Eur. J. Biochem., 101 :395—399 ; (1984); and Liu et a1, Biochem., 18:690-697 (1979)). The modified antibody is then d with the thiol- ning maytansinoid derivative to produce a conjugate. The conjugate can be purified by gel filtration h a Sephadex G25 column or by dialysis or tangential flow filtration. The modified antibodies are treated with the thiol—containing maytansinoid (l to 2 molar equivalent/maleimido group) and antibody- maytansinoid conjugates are purified by gel filtration through a Sephadex G—25 column, chromatography on a ceramic hydroxyapatite , dialysis or tangential flow filtration or a combination of methods thereof. lly, an average of 1-10 maytansinoids per antibody are linked. One method is to modify antibodies with succinimidyl 4-(N—maleimidomethyl)—cyclohexanecarboxylate (SMCC) to introduce maleimido groups followed by reaction of the d antibody with a thiol-containing maytansinoid to give a thioether—linked ate. Again conjugates with 1 to 10 drug molecules per dy molecule result. Maytansinoid conjugates of antibodies, antibody fragments, and other proteins are made in the same way.

In another aspect of the ion, the CD37 dy is linked to the drug via a non— cleavable bond through the intermediacy of a PEG spacer. Suitable crosslinking reagents comprising hydrophilic PEG chains that form linkers between a drug and the anti-CD37 antibody or fragment are also well known in the art, or are commercially available (for example from Quanta Biodesign, , Ohio), Suitable PEG-containing crosslinkers can also be synthesized from commercially available PEGs themselves using rd synthetic chemistry techniques known to one skilled in the art. The drugs can be reacted with bifunctional PEG-containing cross linkers to give compounds of the following formula, Z X1 ( CH2 CHz—O )n Yp D, by methods described in detail in US Patent Publication 20090274713 and in W02009/0134976, which can then react with the cell g agent to provide a conjugate.

Alternatively, the cell binding can be modified with the bifunctional PEG crosslinker to introduce a thiol- reactive group (such as a maleimide or haloacetamide) which can then be d with a thiol-containing maytansinoid to provide a conjugate. In another method, the cell binding can be modified with the bifunctional PEG crosslinker to introduce a thiol moiety which can then be treated with a thiol-reactive maytansinoid (such as a maytansinoid bearing a maleimide or haloacetamide), to provide a conjugate. ingly, another aspect of the present invention is an anti-CD37 antibody drug conjugate of formula (II) or of formula (E'): —CHz—CHZ--»O—),,—Yp—D]m (I?) [D-Yp—(—CH2—CH2—O—)n—X1]m—CB (if) wherein, CB represents an anti-CD37 antibody or fragment; {00184} D represents a drug; ..52..

] X represents an aliphatic, an aromatic or a heterocyclic unit bonded to the cell—binding agent via a thioether bond, an amide bond, a carbamate bond, or an ether bond; Y represents an aliphatic, an aromatic, or a heterocyclic unit bonded to the drug via a covalent bond selected ferom the group consisting of a thioether bond, an amide bond, a carbamate bond, an ether bond, an amine bond, a carbon-carbon bond and a hydrazone bond; 1 is 0 or 1; p is 0 or 1; m is an integer from 2 to 15; and n is an it‘steger from 1 to 2000.

In some embodiments, m is an r from 2 to 8; and In some embodiments, n is an integer from 1 to 24.

] In some embodiments, m is an integer from 2 to 6.

In some embodiments, m is an integer from 3 to 5.

In some embodiments, n is an integer from 2 to 8. Examples of suitable PEG-containing linkers include linkers having an N-succinimidyl ester or osuccinimidyl ester moiety for reaction with the anti—CD37 antibody or fragment thereof, as well as a maleimido— or haloacetyl-based moiety for reaction with the compound. A PEG spacer can be orated into any crosslinker known in the art by the methods described herein.

Many of the linkers disclosed herein are described in detail in US. Patent Publication Nos. 69933 and 74713, and in WO2009/0134976; the contents of which are entirely incorporated herein by reference.

The present invention includes s wherein about 2 to about 8 drug molecules ("drug load"), for example, maytansinoid, are linked to an anti-CD37 antibody or fzagment thereof. "Drug load", as used herein, refers to the number of drug molecules (e.g., a maytansinoid) that can be attached to a cell binding agent (e.g., an anti-CD37 antibody or fragment thereof). In one aspect, the number of drug molecules that can be attached to a cell binding agent can average from 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, 8.1). Nzi—deacetyl-Nzi—(3-mercapt0-l-0x0propyl)- maytansine (DM1) and NT-deacetyl-NT-(4-mercapto—4—methyloxopentyl) maytansine (DM4) can be used.

Tiéius, in one , an congugate comprises 1 sinoid per antibody. in another aspect, an immunocongugate comprises 2 maytansinoids per antibody. In another aspect, an congugate comprises 3 maytansinoids per dy. In another aspect, an immunocongugate comprises 4 maytansinoids per antibody. In another aspect. an immunocongugate comprises 5 maytansinoids per antibody. In another aspect, an immunocongugate comprises 6 maytansinoids per antibody. In another aspect, an immunocongugate ses 7 maytansinoids per antibody. In another aspect, an immunocongugate ses 8 maytansinoids per antibody.

In one aspect, an immunoconjugate comprises about 1 to about 8 maytansinoids per antibody.

In another aspect, an immunoconjugate comprises about 2 to about 7 maytansinoids per antibody. In another aspect, an immunoconjugate comprises about 2 to about 6 maytansinoids per antibody. In r aspect, an conjugate comprises about 2 to about 5 maytansinoids per dy. In another aspect, an immunoconjugate comprises about 3 to about 5 sinoids per antibody. In another aspect, an immunoconjugate comprises about 3 to about 4 maytansinoids per antibody.

] In one aspect, a ition comprising immunoconjugates has an average of 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, 8.1) drug molecules (e.g., maytansinoids) attached per antibody. In one aspect, a composition comprising conjugates has an average of about 1 to about 8 drug molecules (e.g., maytansinoids) per antibody. In one aspect, a ition sing immunoconjugates has an average of about 2 to about 7 drug molecules (e.g., sinoids) per antibody. In one aspect, a composition comprising immunoconjugates has an average of about 2 to about 6 drug molecules (e.g., maytansinoids) per antibody. In one aspect, a composition comprising conjugates has an average of about 2 to about 5 drug molecules (e.g., maytansinoids) per antibody. In one aspect, a composition comprising conjugates has an average of about 3 to about 5 drug molecules (e.g., maytansinoids) per antibody. In one aspect, a composition comprising immunoconjugates has an average of about 3 to about 4 drug molecules (e.g., maytansinoids) per {03201} in one aspect, a composition comprising immunoconjugates has an average of about 2 d: 0.5, about 3 :1: 0.5, about 4 i 0.5, about 5 i 0.5, about 6 I 0.5, about .7 ;+.- 0.5, or about 8 i 0.5 drug molecules (e.g., maytansinoids) attached per antibody. In one aspect, a composition comprising immunoconjugates has an average of about 3.5 2+: 0.5 drug molecuies (e.g., maytansinoids) per antibody. {0902025 The anti4CD37 antibody or fragment thereof can be modified by ng a. tional crosslinbing reagent with the anti—CD37 dy or fragment thereof, thereby resulting in the covalent attachment of a linker molecule to tbe anti~CD37 antibody or fragment thereof. As used herein, a "bifunctional crosslinking reagent" is any chemical moiety that covalently links a cell—binding agent to a drug, such as the drugs described herein. In another method, a portion of the linking moiety is provided by the drug. In this respect, the drug comprises a linking moiety that is part of a larger linker molecule that is used to join the cell-binding agent to the drug. For example, to form the maytansinoid DMl, the side chain at the C—3 hydroxyl group of maytansine is modified to have a free sulfliydryl group (SH). This thiolated form of maytansine can react with a d cell—binding agent to form a conjugate. Therefore, the final linker is assembled from two components, one of which is provided by the crosslinking reagent, while the other is provided by the side chain from DMl.

The drug molecules can also be linked to the dy molecules through an intermediary carrier le such as serum albumin.

As used herein, the expression "linked to a cell—binding agent" or "linked to an anti—CD37 antibody or fragmen " refers to the conjugate molecule comprising at least one drug derivative bound to a cell-binding agent anti-CD37 antibody or fragment via a suitable linking group, or a precursor thereof.

One g group is SMCC.

In certain embodiments, xic agents useful in the present invention are maytansinoids and maytansinoid analogs. es of suitable maytansinoids include esters of maytansinol and maytansinol analogs. Included are any drugs that inhibit microtubule formation and that are highly toxic to mammalian cells, as are maytansinol and maytansinol analogs.

Examples of suitable maytansinol esters include those having a modified aromatic ring and those having modifications at other positions. Such suitable maytansinoids are disclosed in US. Patent Nos. 4,424,219; 4,256,746; 4,294,757; 4,307,016; 946; 4,315,929; 4,331,598; 4,361,650; 4,362,663; 4,364,866; 4,450,254; 4,322,348; 4,371,533; 5,208,020; 5,416,064; 5,475,092; 499; 5,846,545; 6,333,410; 7,276,497 and 7,473,796.

In a certain embodiment, the immunoconjugates of the invention utilize the thiol-containing maytansinoid (DMI), formally termed Nzi—deacetyl-NZZB—mercapto—1- oxopropy1)-maytansine, as the cytotoxic agent. DMl is represented by the following structural formula (111): o 91% T SH 9' \ <3 «"3. - 9x, ;« , x. - w , K .V- \ .14 \ .2- \Y \x‘ \xE-e" ' \rA:‘~.- U l K , l K , :, t 1 .=: t " 4, -\,\~/ \ ~"flA\‘L‘O_ \k, ,‘v"\\ /‘:\, \Nfi/I/ .v" N, & ‘,\" \' H" ‘ N H I O MeO (111) In another embodiment, the conjugates of the t invention e the thiol-containing sinoid acetyl-N27(4—methy1mercapto—l- ty1)—maytansine (e.g., DM4) as the cytotoxic agent. DM4 is represented by the following structural formula (IV): 23' \ 2: 2 C. 2222*]wa \2 3 2222\2 wa/‘l «(g-"Lu, 2, y"\\ %‘\ i \‘fig «~+:L\ i S C N 0 M90 HO H (IV) Another maytansinoid comprising a side chain that contains a sterically hindered thiol bond is N2,~deacetyl-N-2’(4-mercapto-l—oxopentyl)—maytansine (termed DM3), represented by the following structural formula (V): Meé (V) Each of the maytansinoids taught in US Patent No. 5,208,020 and 7,276,497, can also be used in the conjugate of the present invention. In this regard, the entire disclosure of 5,208,020 and 7,276,697 is orated herein by reference.

Many positions on maytansinoids can serve as the position to chemically link the linking moiety. For example, the C—3 position having a hydroxyl group, the C-14 on modified with hydroxymethyl, the C-15 position modified with hydroxy and the C—20 on having a y group are all expected to be useful. In some embodiments, the C-3 position serves as the position to chemically link the linking moiety, and in some particular embodiments, the C-3 position of sinol serves as the on to chemically link the linking moiety.

Structural representations of some conjugates are shown below: 2"".

I.,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, Ab = Antibody = H or Me (V1.3 9.». N: \ n... \f \w‘ f. E .f/ O 1 ‘ 'Q i ‘ i _,r‘\\ s 0 Ab = Antibody Ab—PEG4-Ma1—DM1 (VII) 0%W O N S 0‘ , 3‘, \\ ‘Y’ c’"'\o Ab = Antibody R' = H or Me DM1: R=H, q=1 DM4: R= CH3, q=2 = 1-24 \ ".\ 4 5° 00 AbPEG—SIA—DMl/DM4 (VIII) -57..

....... M 2 Ammo /x4 1m, wwmx .¢\\O H‘50 NHA0 Ak."SMCODM1 (1X) \ 0 O .I \ Hum Mn.0 is \ Null go,V\«§§$L..m:i§ $«.A\ \N OHHW O__._\ 1.

V xxx"m a. \II. \\ fall, \xx. xx .\{Ytz‘xxx r: O Ti? Nu/,A,\ m:W IA H550 E M Ab: Antibody \- J 2—5 —DMI (X) S ,.\ Oufiuk / x x k w S \Eiithz V3111}. x, wAb Ab Antibody Ab-SPP—DMl (X1) a" \ t O S \ \\ , O O 9' \ Et ? . a 3‘ .. 1.7-» w MeO\/\ N \,./\,I"§ \a \\\\ U\5J ,L l Ab—A_ m1'b d0 y ""aim/"kl,..«""\,\_,,_..._i>\, x I s‘ é ‘N O MeO Ho H ~, 2-5 Ab—SPDB-DM4 (XII) x "\ §~ $O§Na+ § 0 i :H VT S\S "Axxx ! \xvéNMAb, r’ ‘O "We, ~\_ /_.a\ i k ibody § V" ‘v-ws‘x 3 \S 5 N O : ‘ MeO HO H "a, "9"" Ab-SulfO-SPDB—DM4 (XIII) Several descriptions for producing such antibody—maytansinoid conjugates are provided in US. Patent Nos. 6,333,410, 6,441,163, 821, and 7,368,565, each of which is incorporated herein in its entirety.

In general, a solution of an antibody in aqueous buffer can be incubated with a molar excess of maytansinoids having a disulfide moiety that bears a reactive group. The reaction e can be quenched by addition of excess amine (such as ethanolamine, taurine, etc). The maytansinoid-antibody conjugate can then be purified by gel filtration.

The number of sinoid molecules bound per antibody molecule can be determined by measuring ophotometrically the ratio of the ance at 252 nm and 280 nm. The average number of maytansinoid molecules/antibody can be, for example, about 1—10, 2-5, 3-4, or about 3.5. In one aspect, the average number of maytansinoid molecules/antibody is about 3.5 i 0.5.

Anthracycline compounds, as well as derivatives, intermediates and modified versions thereof, can also be used to prepare anti-CD37 immunoconjugates. For example, doxorubicin, doxorubicin derivatives, doxorubicin ediates, and modified doxorubicins can be used in anti-CD37 conjugates. Exemplary compounds are described in , which is herein incorporated by reference in its entirety. Such compounds include, for example, compounds of the following formula: wherein R1 is a hydrogen atom, hydroxy or methoxy group and R2 is a C1-C5 alkoxy group, or a pharmaceutically acceptable salt f.

Conjugates of antibodies with maytansinoid or other drugs can be evaluated for their ability to suppress proliferation of various unwanted cell lines in vitro. For example, cell lines such as the human lymphoma cell line Daudi and the human lymphoma cell line Ramos, can easily be used for the assessment of cytotoxicity of these compounds. Cells to be evaluated can be exposed to the nds for 4 to 5 days and the surviving ons of cells measured in direct assays by known methods. IC50 values can then be calculated from the results of the assays.

The immunoconjugates can, ing to some embodiments described herein, be internalized into cells. The immunocongugate, therefore, can exert a eutic effect when it is taken up by, or internalized, by a CD37—expressing cell. In some particular embodiments, the immunoconjugate comprises an antibody, antibody fragment, or polypeptide, linked to a cytotoxic agent by a ble linker, and the cytotoxic agent is cleaved from the antibody, antibody fragment, or polypeptide, wherein it is internalized by a CD37-expressing cell.

In some ments, the immunoconjugates are capable of depleting B-cells, e.g. autoreactive B-cells. For example, in some embodiments, treatment with an immunoconjugate results in a depletion of at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, or at least about 75% of B-cells.

In r aspect of the invention siRNA molecules can be linked to the dies of the present invention instead of a drug. siRNAs can be linked to the antibodies of the present invention by methods commonly used for the modification of oligonucleotides (see, for example, US Patent Publications 20050107325 and 20070213292). Thus the siRNA in its 3’ or 5’—phosphoromidite form can be d with one end of the crosslinker bearing a hydroxyl functionality to give an ester bond between the siRNA and the crosslinker. Similarly reaction of the siRNA oramidite with a crosslinker bearing a terminal amino group results in linkage of the crosslinker to the siRNA through an amine.

Alternatively, the siRNA can be derivatized by standard chemical s to introduce a thiol group.

This thiol-containing siRNA can be reacted with an antibody, that has been d to uce an active disulfide or maleimide moiety, to produce a cleavable or non cleavable conjugate. Between 1 — 20 siRNA molecules can be linked to an dy by this .

III. Polynucleotides In certain embodiments, the invention encompasses polynucleotides comprising polynucleotides that encode a polypeptide that specifically binds CD37 or a fragment of such a polypeptide. For example, the invention es a polynucleotide comprising a nucleic acid sequence that encodes an antibody to a human CD37 or encodes a fragment of such an antibody. The polynucleotides of the invention can be in the form of RNA or in the form of DNA. DNA includes cDNA, genomic DNA, and synthetic DNA; and can be double—stranded or single—stranded, and if single stranded can be the coding strand or non-coding (anti-sense) strand.

In certain embodiments, the polynucleotides are isolated. In certain embodiments, the cleotides are substantially pure.

The invention provides a polynucleotide comprising a polynucleotide encoding a polypeptide comprising a sequence selected from the group consisting of SEQ ID NOsz4—l20.

The invention further provides a polynucleotide comprising a sequence selected from those shown in Tables 7—10 below.

Table 7: le heavy chain polynucleotide sequences mmVHPolvnucleotide Sequence(SEQ_:IDNO) ‘""w"m"" § n1UCH)37-3 i 3°gtgggégtéagésagtcaggacctggcctggtggcgccctcacagagcctgtccanacaxgcactg tctcagggttctcattaaccacctctggtgtaagctgggttcgccagcctccaggaaagggtctggagtg gctgggagtaatatggggtgacgggagcacaaactatcattcagctctcaaatccagactgagcatcaag aaggatcactccaagagccaagttttcttaaaactgaacagtctgcaaactgatgacacagccacgtact Iactgtgccaaaggaggctactcgttggctcactggggccaagggactctggtcacagtctctgca (SEQ IDNO: 121) {fichCD373 aagcttgccaccatggctgtcctggcactgctcctctgcctggtgacatacccaagctgtgtcctatcacaggtgcaggtg aaggagtcaggacctggcctggtggcgccctcacagagcctgtccattacatgcactgtctcagggttctcattaaccac ctctggtgtaagctgggttcgccagcctccaggaaagggtctggagtggctgggagtaatatggggtgacgggagcac . tcattcagctctcaaatccagactgagcatcaagaaggatcactccaagagccaagttttcttaaaactgaacagtz? ctgcaaactgatgacacagccacgtactactgtgccaaaggaggctactcgttggctcactggggccaagggactctggg itcacautctct*cagcctctacvaam‘wccc(SEQIDNQ: 122) huCD37-3v10 aagcttgccaccatgggttggagctgcattattctgtttctggtggccaccgccaccggtgtgcactcacaagtccaagtc 1"" caagaatctggtccaggtctggtggccccttcccaaactctgagcatcacctgtaccgtttctggttttagccttaccacctc itggtgtgagttgggtacgccaaccacccggtaagggtctcgaatggctgggtgtaatctggggtgatggttccacaaatt accatccttccctcaagtcccgccttagcatcaaaaaggatcacagcaaaagtcaagttttcctgaaactgaatagtctgac agcagccgatacagccacctactattgcgccaagggtggttatagtcttgcacactggggtcaaggtaccctcgttaccgt: umnniflctcctcactagtaccaaz.fgeccc(SEQIDNQ:I23} __ j t huCD37—3vll gccaccatgggctggagctgtatcattctgtttctggtggcgacagctactggggtccactcccaagtgcaggta tccgggcctggattggtcgcaccaagccagaccctctctatcacttgtaccgttagcgggttctctctgacaacc Gtgfaztgagttgggtga=x(cagccaccaggaaagggactxtiagtggctgggggtgatttg 2* g : LaaactatcaitccagtcfiaaatL’nggtLGtLLattaaz.c1d&z,dCCZ€idLId.idfCtCfiflngL1' aaactcaaLagLL. gaaagccOcafiacactgaacgafiactfiLgLLaaaHUaLL sjaLLLivtLaiLLgcatcazicaaa L.c{SEQ 113‘ N(J:1 "’ V..LLLL.L"....... 777777....LLLLLLLLL"""".._. 5..."...«n 12 EcagamcagiL‘tggtg agtctggacm agdagLLt‘ggdgq(magma... U Lfictgggta‘tacattcacaaagtatggaaigaactgggtgaagcaggct'aamadawvtfiaaagtg L'tga"{*aafiLngggdLagrtttgcctic 1 gatgggctggataaacaccaacactggagagtcaagaaatgc 3 tctflggaaacctctgccacvcactrILLi‘att‘LQcaLzatLaaLaLLLEL:aaatatgaggacacggctacat nggaaggggL-LgtagLLgLLoaLtggngcaaLLdLLaciLtLaLagtLtLLLca (SEQID 3.....L....,,_............................ . .1 VVV........ ....LL.LLL..LL........

V........... .LL.............._._................ gchCD37- 12 aagcttgc3LLatggg-aogiLaioca‘aaiLLLL'tiLtgotcgctaLtuLmLLggt9:91.LtLaLaLaitLaa1‘91; ttcaaagiggccmoathbaoaaagLagggaaaoatwaaadtaathLLaaggca mggfiacactficacaaa cai(53.81;{9.331033%amcccagggcaagf "{LLLadana‘iga0'{Egg}3'9:LaaiaccaacactcDgazg agtLtL-iwaatLLigavgaomaaggpccwffloficwcdggagaca'w gccagcac.LagLLfiacctgcaaafi aaLaatLtgaagtatgaEgltdgdgfldfiLCLfimGEGCgELNngLac-tgLLgfigLLagactggggacaaggtacca ‘ act);m‘cLaLLva3:caL{$12.00LL (QLQ 11) N(J:126}"3v" "W.L........._.‘"............ ... VV.V.....................L L.,a Digcagm4337335333,:«Ctgat.CtgéfigdadcfimkdfitLLttLLaLmaLLLgL (3O teactggctactccatoaccagtgamiggctggcac.tggaicLLL"castfiLLaOLaaaaagctga3'0 atggatggcctacaiacmiacagig.{giggcacto6LtaLaaLLLathLtLama‘rtLULLatL‘"Ltac aLILLvagacaLfi‘" acaaccam‘tLttLLLgLfiofigfik1gt£VaLv1chiJaz§E~Y’lcaC&gLcacai afiactgtgcagdrwcLaLtaLULUacgg{VLLtogmUmscwgwcLaaggactctggmac l"i?§s:33f:§.<.§fl;11}.3§91" _...- _________________________________________________ «any...,,,,,,,,H,..",,......,,..................................___.._chCD3738 E aagcfigccaccatg gciggag‘itgiatcafictgmfio3190mmLGLLLLLiWaLLLLatLLLLLaotLLaaLtLL aggaatctggccm‘gacctacttaaUL‘VLiLLcagaO'cLILrccctoaLLTgLameLagbamLicaatca'L‘aiLLag octfigt‘Ltgficac‘woatcasmaatitLLLgoLaacaagttggviatogatgthiacafictgtatagngjgggtaccg 5 1:) attacm L‘Li Lthcaaga9ccgaaL‘zMatLaLLai—Jf(rataLawLacLLaa-‘caamfitctm QLLtcag{(2:ngt: acia"°ga.dg..3.iacgcta.taLtattgtPLLagmLtaLtaiggaiat’rgfiLdtLguLOELLLthggggccaggga acccm‘tagggg34mg. ' V a ' v; ;, 33) . .7.....L...LLLLL"3.""....L..... hUCB37'38 aagcétgcc' "3&th,gtigga'5 ctggzLiLLdLiLaLaogtLLangc aagavtLLguicgmguggaaacccaocLaLtLLcTL.gtLtLaLLLLmeiLt.LLigthaLtLtmaLcagtgg1&1"). 0{toggLtgvcafiggmmggwg‘mcLL‘9«{taagvggctggagto93*ggbLatatatLLt9faLaUL3332123933"c gai‘tacaaLtcL.agtctgaayaficafiga{€304.4thcQLLLLaLawLaaaaaccafit’rmLLnogthgCtsigi . : 5E?.3.’*d0LtgagaLaLchLtaLnaLtafiotngtggmLidtgngtowLLIggmgtgtafivqggacaaw ’ ‘ ' ‘ "c @331955‘foHJNQ: 179) _ » -_. muCD37-50 ag gLagLfiLagg gt aggucdgacLiofiganLLtttLaJLLdLmLaLtLaLctcactg {LaLWOLtaL:CLthcaLLagtagmchtcC,L'1L£6gatLL69agttmaggaaacaaaaégga atggatggga.taLaLaLtL{110391353918gLaLtgthLagLLLaitctcaaaagt-c.gaatctctam actegagacactLLaaLaaLLaLLtLLiLLLLLagttoaartL-Lfih'acgamaLaLagLLde‘:(O g .......... afiaLtngaagwggmLLaivgitaL‘LLLgcLtggmgcfiactggggccaagggaLLtciggtLac L.:3:3:ct:,_..:ca<83" .."a"",.V,..,..,,,,..‘,.........................................'Hii‘éfié'ifimw ELCLlLLc‘l‘nggbgig’kaLOLataatLLthLt gtigciactctaccvcavtccafi'VL'wggtgcagctgL figgagtccggCCCCi‘flOL-EULiCddUL‘LfiL-RagagtLtgaotrgacfiEL‘LEgmLWLLMLaGLtagccagcg gmcgcfiggLaLfgLLfiLagaLducatLLwLLaaLaaacgflagtogatog&atacdmtLtaLE’LaLOcLLaaLL g:ctattchthcLLtgaaatnccggatcacmiaLccgtL33:10acmtaagaas.LaWLtQLachtgaaLaflLvtt' L<{Lagugacamocam013k1:81.{EEGCLngjLUd'LéifldU’gLitnggdgcfigvfLthtacthggLLad90 ' 5CflCC-G muCD3751 gatgigcagLtléflggfigtLfiboakabchtgfiammHLLLaL‘LLtuLaLiLa.LEgLaLtg3 gciacLieaLLLLLangmLOLLEggLacfggamggagiticcaggaaacaaadgga Egg,thaLstaLaLtdLaUELLagL-aLtaaL‘taLaocLLamNcaaaagtcgaatctctam actsg2agactcatLcaagaaLLagficficctmaE,ttLaaLtLtLLLaLtdLigaagaLacagccacat E gcaagvawaiaaatgvgmcngLLt9.j€ttLina.[Ugggccaagggactciggtcac "L La ssmmxo 133) AagcttgLLaLLaLngLLLLLLfiLLatLaLLLtgmLtggtggccactgwacirm " LLLaOGaaichf-L:1321""C(- try! (n \ , »'\ L- 4 (L ., ,L -62— gctttgcttggcactggattcggcagtttccaggcaagggactogaatggatgggctacatccattacagtggctcaac caattacagccctagcctgcagggccgaatctctattaccagggatagttctattaaccagtttttcctgcagcttaattccgt gactgcctctgacacagcaacttactattgcgcecotggctactacoggttcOgagcctgtttgtatactggggtcaggg muCD3756 gatgtgcagcttcaggagtcaggacctgacctogtgaaaccttctcagtcactttcactcacctgcactg tcactggctactccatcaccagtggttttgcctggcactggatccggcagtttocaggaaacaaactgga gggctacatacactacagtggtggcactaactacaacccatctctcaaaagtcgagtctctatc 1 actcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaa gaggctactatggtttcggggcctgg’cttgcttactggggccaagggactctggtccc tctctca{SEQ IDNO132) huCD37—56 aagcttgccaccatggagtggagctgcattatcctgttcctcgtcgccaccgcaaocggcgtccactcccaggtgcagct gcaagaaagcgggccaggattggtaaaaccttcccagtctctgagtcttacttgtaccgtatctggatacagtatcacatct ggcttcgcctggcattggattcgccagtttcccggcaaggggcttgagtggatggggtatattcattattctggaggtacca actacaacccttccctgaagagtcgagtctcaattaccagggacacttccaagaaccaattctttttgcagcttaattcagtg gccgacaccgctacttactactgcgcccggggctactatgggtttggtgcctggttcgcctactggggccaggg [NEWA‘.1gatgtgcagcttcaggagtcaggacctgacctgfigaaaccttctcaotcactttcactcacctgcactg tcactggctactccateaccagtggttttgcctggcactggatocggcagtttccaggaaacaaactgga atggatgggctacatactctacagtggtagcactgtctacagcccatctctcaaaagtcgaatctctatc .: actcgagacacatccaagaaccagttcttcctgcagttgaattctgtgactactgaggacacagccacatattactgtgcaa gagggtactatggttacggcgcctggtttgcttactggggccaagggactctggtcactgtctetgca (SEQ ID 311L114) huCD37—57 aagcttgccaccatgggctg0agctgcatcattctgtttctggtggccacagcaactggcgttcacagtcaagtccaactg caggagagcggccccggactcctgaaaccatctcagtcactcagtctgacatgtactgtgagcggctacagcattacctc aggcttcgcttggcattggatcaggcagttccccggaaaaggtctggagtggatggggtacattctgtacagcggcagta cagtgtattcaccctccttgaaatctaggatatcaatcacacgtgatacaagcaaaaatcagttcttcctccagctgaactcc gtcaccgccgcagacacagcaacctattattgtgctcgcggatactacggatatggcgcatgttcgcctattggggcca ggggacactcfituaccgtttccrgccocctccacaaam:Hccc(SEQIDNQ:135__)_ 2523 caggtggtggagtctgggggagacttagtgaagcctogagggtccctgaaactctcctotgcagcctctggat tcactttcagtagctatggcatgtcttgggttcgccagactccagacaagaggctggagtgggtcgcaaccattagtagtgz:\ gtggtagttacacctactctccagacagtgtgaaggggcgattcaccatctccagagacaatgccaagaaaaccctgtac ctgcaaatgagcagtctgaagtctgaggacacagccatgtattactgtgcaagacatagttactacgatactagcgtcgac tactggggtcaagaacmiségcgtctccwMSEQlDNO182) 1 Table 8: Variable light chain polynucleotide sequences EmuCD373 gacatccagatgactcagtctccagcctccctttctgtatctgtgggagaaactgtcaccatcacatgtc ~ gtgagaatattcgcagtaatttagcatggtatcagcagaaacagggaaaatctcctcagctcct ggtcaatgttgcaacaaacttagcagatggtgtgccatcaaggttcagtggcagtggatcaggcacacag tattccctcaagatcaacagcctgcagtctgaagattttgggacttattactgtcaacattattggggta "E"CtaCi1aszacwttcggtggaggcaccaa)c ‘aatcaaac3t(SEQ1J2NQ:136) 63133737" gaattcgccaccatgagtgtgcccactcagg, Ctggggttgctgctgctgtggcttacagatgccagatgtgacatccag atgactcagtctccagcctccctttctgtatctgtgggagaaactgtcaccatcacatgtcgagcaagtgagaatattcgca gtaatttagcatggtatcagcagaaacagggaaaatctcctcagctcctggtcaatgfigcaacaaacttagcagatggtgt s gccatcaaggttcagtggcagtggatcaggcacacagtattccctcaagatcaacagcctgcagtctgaagattttggga E cttattactgtcaacattattggggtactacgtggacgttcggtggaggcaccaagctggaaatcaaacgtacg (SEQ ‘ 11DNO:137) huCD37—3 gaattcgccaccatgggttggtcctgcatcatcttg‘tttctcgtggccacagccaccggtgttcactctgatatacaaatgac (1.0 and 1.1) tcaaagcccttccagtttgagcgtaagtgtgggtgaacgcgtaacaatcacctgtaoagctagtgaaaacatccgcagta 1 M55111".‘1...~...."mv....w$3.». .....

T1,",Mflflflflwu. cttcactattctctggttcaaottcg g_maccGattattcacttaao’ttcaactcacc:t accagaacatttcgcctacatatta ictgtcaacaactactgg«4.1cgtacgacctggacattcggtcaagrtactaagccggaaatcaagcgtacg {SEQ 113 WM) 1 muCD37 12 gt ctaacacaccctcctgc151;cttaotgtatctctg fiwccc'wccaaactcagta‘a tacgatact‘atttgtactggttccavcao'Laaaggacagcc fiacccaaactcficatcaactatg‘icaacxtamatctggggtca'V°caggttcagtmcact{2g tctgggacagacttcaa‘cccaacatcatcctotggaggaooagcatactgcaacatattacwtcaac ' " iacac‘1t0_gaooattccgtacacattcggacgcgacmazctggaaataaaas ...........................g. chCD37-12 oaattcoccaccatgc"ttgvtcctgtamacctOEtctcggtgcancccractgccgmatactgatattctacis:act005 gtcaccagccagtctcficamgtccct00occ'tgcgtgcaccatcccctccccogcctcacagtccUtgagcac1a attcctatctctactgctttcaacagaagccawaagcc‘ctacgctgctvatcaaotacnctccaacctcgc caccgmcttcccgctagattctctggttccggtagcUgaact’satttcactttgaacatccacccccttgaggaaagga tacc ccacttacéattctcaacactcttgaoacattccttaacctttgcaggaogaacaaagctccaaattaaacctacg 1315ch . .». 1 muCD37—3 8 caaattgttctcacmagtctccagcaatcatgtctocatctccacgggacaagotctcacctgca gtgccagct‘aao'tgtaacttacatmactovtaccaagaagtcawcacct aaaagatggat ttatgacacatccaaactcocttctggaotccctmtcccttcactYfY’ggtUggtcgggaacctcttac tctctcacaatcaocascatggaccctcaacatcctcccacttattac1gccagcagtgP‘xz rgattactaam i ‘ cacccac: .tcc ‘ c’vgaccaagctgcaaattaaacc) (SEQ 11) ___..u.._............""......____________________ "c" \<""" chC1337-38 ; gcgctggtcctotattacctgtttctcctgcccacaociataggtgttcattctcaJ L .ttgtgctgac ccaatcaccavctattatvtccgtagcccggccagaaagagacaatfiacatctaowcta01tcttctgtgacttacat gcattggtatcaacagaagtcaggtaccagtcttaaacgttooatcwccacacatccaaactcxtcutccg.. CC? tg ccaggficagcggaggtgggtc.ccaccagttamac'gaccatatcctct'ttgmacctcaacawctoctacitatta ttgtcaacaatggatttctaaccccccccaccttiggtggngaacaaagctggagatcaagcgtacg (SEQ ID N(3:142) : gaattcgccaccatccvv{gotcctgcattattctgttcttggtcgccacto1tactcccgacactctattctvctcacaCy "unannflflflrflwr . E c 121510311"de(XLfingCCLUCt'CCCCGQfi’fgaoc(milk*c'ttgtcatoctctoccagttcctcg:.gacatatatgc ~ attwtatcaccaaaaacggtacctctccaaaaaatggatcctacgacacttcaaagcttowtaggcgttcctgcca t gattttccgggtctgggictggcacttcaacagtctgacccttwtt’catgaancivaagacgca‘ccacctattactgt t .\ t cagcagtggatttcaaatctccaccitcggcggcgyaacaaactggagataaagcgtacv {8131:1113 ‘ w NO: 143, caaattcttctcacccagttccagcaacatgtciocatctxcac.g ggacaaggtcaccatgacdgca gtgccacctcaagtgtgacttacatgcactgstaccagcagaagtcagg;acctcccccaaaagatggatuatgaca"> 1,.V......_________ [ccaaactgccttatgagtccxtggtcgtttcactggtacgggtctgggattactctctcacaatcagcagcatgg antarctoaagatoctoccacttattactgccaocao‘rgwagtoataacccccccacgttcggctcggggacaaagttgga {3EQMID N01143: Wmflflfl,flflfl,",, ccaccatgccacfi"‘atrtcattattctctr‘tugttgctacgmaggagtacatag oasaagtccicac gcattcgtatcaocaaaagcct"occaatcccctaaaagctgga‘tctaccatactttta'atccocataccctgtgcccgc aafmttctcongagtgfigcagtogagcacrtatagtctcaccatcactiaatggaagtagaogtgcagcaacctatt acctcaocaotgvt"gataatcccgctacttttggtcagggtacaaaficumwattaagcctac ( ‘EEQ 1D 0:1/1_‘?_:__ muCD37-5 1 caaattgttctcacccagtctccagcaatcatgtctgcatctccaggggagaaggtcaccatgacctgca gtgccacctcaagtgtgacttacatgcactggtaccagcagaagtcaggcacctcccccaaaagatggatttatgacaca tccaaactggcttctggagtccctgctcgcttcagtggcagtgggtctgggacctcttactctctcacaatcagcaacatgg aggctgaagatgctgccacttattactgccagcagtggagtagtaacccacccacgttcggctcggggacaaagttgga am(SEQID149146),,,,,,,,,,,,,,WM 1111163637351 . t gaattcgccaccatgggatggagctgtattattctgttcctggttgctactgctactggcgtccattccgagatagtcctcac t ccagagccccgcaaccatgagtgcctcccctggggagcgagtgactatgacttgttccgccacttcttcagttacctatat \ gcattggtatcagcagaaacctggacagtctccaaagcgttggatttacgacacctccaacctggcttcaggagttcctgc 1 taggttcagcggatctgggtctggcacaagttattcactcaccattagttccatggaggccgaagatgccgctacttactac 1 tgtcagcagtggagcagcaacccccctacattcgggcagggaactaagctggagatcaaacgtacg (SEQ ID Lcccap—Magcauuu. ........"v. ___NO:14_7) muCD3756¢ caaattgttctcacccagtctccagcattcatgtctgcatctcCaggggataaggtcaccat9acct9ca gtgccagttcaagtg?acttacatgcactggtatcagcagaagtcaggcaoctcccccaaaagatggatttatgacacat CCaaactggcttctggagtccctgctcgcttcagtggcggtgggtctgggacctcttac tctctcacaatcagcaccatggaggctgaagatgctgccacttattactgccagcagtggattagtgacc EECacccaCOTtCMaggg ggaccaaoctggaaataaaacm(SEQIQNQ:__148) ..

E huCD3756 gaattcgccaccatgggctggtcct cctgtttctggtggcaaccgctactggg ttcaCECtatattgtcctgacg g g E : E acagagtccagccttcatgagtgcttctcccggagaaaaggtcacaatgacttgttcagcttcctcctccgtcacatacatg E cattggtaccagcagaagcCtgaccagagtcctaagaggtggatctatgatacaagcaatctggcttccggtgtcccctc E ccgcttttcaggcggcggaagcggaactgactatagcCttaccatctcctcaatggaagccgaggacgctgctacatatt E E actgccagcaatggatcagcgaccctcctactttcggacagggaacaaaattggaaattaagcgtacg (SEQ ID muCD37—57 caaatt9ttctcaccCagtctccageaatCatgtctgcatctccaggggagaaggtcaccatgaCCtgca Egtgccacctcaagtgtgacttacatgcactggtaccageagaagtcaggcacctcocccaaaagatggatttatgacaca tCcaaactggcttctggagtccctchgcttCagtggcagtgggtctgggacctcttactotctcacaatcagcagcatggE Eaggctgaagatgctgccacttattactgccagcagtggagtgataacccacccacgttcggctcggggacaaagttgga E aataaa‘CmfSEQ ID NO:150) E huCD3 7-57 gaattcgccacCatggggtggtcctgtattatcctgttcctggtcgcaaccgccacaggcgttcactccgagatcgtgttga 1 ctcagagcccagccaCCatgtCCgcttCCCCCggggagagagtgacaatgacttgttccgccacaagttctgtaacctac atgcattggtaccagcaaaaaccaggacagagtccccgtcgttggatttatgatacctctaacctggcttcaggcgttcctgE CCcgcttttctggtagtggatctgggacttcctatagccttaccataagctctatggaagccgaggacgccgctacatacta ctgccagcagtggagtgataacccccccaccttcgggcagggaaccaaattggagatcaaacgtacg (SEQ ID No912 _I. 252-3 E gatatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttmagggc 53 aagtcaggacattagcaattatttaaactggtatcagcagaaacCCgatggaactg’ctaaactcctgatctactac acatcaaaattacactcaggagtcccatcaaggttCagtggCagtgggtctggaacagattattctctcaccatt agcaacctggagcaagaagatattgcCacttacttttgccaacagggtaatgcgcttccgtggacgttcggtggE caa9ctaactcaaacgg (SEQ ID NQ: 183) Table 9: Full-length heavy Chain polynucleotide sequences Antlbodx ~ HeaV ham Pol‘nucleotlde SC}:EuenCC (SEQID NO‘ E ___." __ __enfth ChCD373 aagctthCaccatggctgtcctggcactgctcctctgcctggtgacataCCCaaoctgtgtcctatcacaggtgcaggtg aaggagtcaggacCtggcctggtggcgCCCtcacagagcctgtccattacatgcactgtctcagggttctcattaaccac ctctggtgtaagctgggttcgcCagcctccaggaaagggtctggagtggctgggagtaatatggggtgacgggagcac aaactatcattcagctctcaaatCCagactgagcatcaagaaggatcactccaagagccaagttttcttaaaactgaacagt ctgcaaactgatgacacagccacgtactactgtgccaaaggaggctactcgttggctcactggggccaagggactctgg E tcacagtctctgcagcctctacgaagggcccatcagttttccccttggctccaagttctaaatccacaagcggtggaacag Ctgcactgggatgcctcgttaaagattatttccctgagcctgtgacagtgagctggaatagcggagcattgacttcaggtgt gCacactt‘ttcccgctgtgttgcagtCctccggtctgtadcactgtccagtgtcgtaaccgtcccttctagcagcttgggaa CCcagaCctacatctgtaacgtcaaccataaaCCatccaacacaaaggtggataagaaggttgaaccaaagagctgtga acatacatgccctccttgtcctgcaccagagctcctcggaggtccatctgtgttcCtgtttccccccaaacccaag gacactcttatgatctctcgtactccagaggtcacctgtgttgttgtcgacgtgagccatgaagatcccgaggttaaattcaa CgtggatggagtcgaggttcacaathCaagaCcaagcccagggaggagcaatataattctacatatcgggta gtgagcgttctgaccgtgctcCaccaagattggctcaatggaaaagagtacaagtgcaaggtgtccaaeaaggctcttcC cgctcccattgagaaaactatctccaaagccaaggggcagccacgggaaccccaggtgtatacattgcccccatctaga gacgagctgaccaagaaccaggtgagtctcacttgtctggtcaaggggttttacccttctgacattgctgtagagtgggag tctaacggacagccagaaaacaactacaagacaactcccccagtgctggacagcgacgggagcttcttcctctactcca agttgactgtagacaagtctagatggcagcaaggaaacgttttctcctgctcagtaathatgaggctctgcacaatcacta iaéf 1ggtgtg§o[0'1[1: 1ss1:15gtstgkfiggsss1fisssaaaststriavatsasatgtasgstttst._, ..agssttassascts fl? ::ggmwsassa1sssgtaagggttsaaigfisugs'gtsattogggtoatgg‘tssasaaatt assatsattssstsaagtsssgssuagsatsaaaaagga1sasags:1-aagtsaautttss10gaaastgaatagtstgas assagssgatasaassasstas.a1tg1.sssaaoLstmt‘tataotstwsasas111<5<5g1saags1asastsgttassg... s1s13mg11,1.11s1agwa102119210111113:ssttgoatssaautstaa.tssasaagsUgtggaasagagsast ggga112:1.sx11taaag411atttssstgagsstgtgasagtgagstggaamgaggagsaflgasttsaggtgtgsasas ttttss1ssgstgtgttgsagtastsaggtstgtastaastgtssagtgtsgtaassgtsss11stagsagst1gggaasssaga 01.11.112.111.1g1aasgtsaassataaassatsaaasaaaa:ggtggataagaav.10111121211:"aaa{41101110111111.1121gas 210111111:a1gsss1ssttgtss1gsassagagstssts gtssatstgtgttss1st1tssssssaaasasaaggasast stta102111:1stagtastsaagaggtsass{gtgttgfi111:sgzst.gtgagssatgaagatssagdggtwaattsaastggtax. 1.Utggatggagtsgaogttcasaatgssaagasaagssssagggaggagsaatataattstasatatsgggtagtgaga 0Hsf1711CLW1TLELCSLLth_211tggstsaatggaaaagagtasaagt£1121aggtgtssaasaaggstsasssgstss attoogaaaasmtstssaaasssaagg.aossasggcaassmf5gtg121121132111gsssssatstagagaaga r1aigassaaoaassaggtvagtswzsagtstgmaggggttttasssttst \\\\\\\\\\\\\\\\\\\\\\\\\\Wu—hw aggagtccggccccggcctgctcaagccttctcagagtctgagtctgacttgtactgtttctggctacagcataaccagcg gtttcgcttggcactggatcagacagcatcccggcaacaaactggagtggatgggatacatactgtactcaggctcaact gtctattccccctccctgaaatcccggatcagtattacccgtgacacttctaagaaccatttttttctgcagctgaacagcgtt accgcagctgacactgcaacctactactgtgcccggggatattatggatacggagcttggttcgcttactggggccaagg caccctcgtaactgtgagtgctgcttccaccaagggcccatcagttttccccttggctccaagttctaaatccacaagcggt ggaacagctgcactgggatgcctcgttaaagattatttccctgagcctgtgacagtgagctggaatagcggagcattgact Etcaggtgtgcacacttttcccgctgtgttgcagtcctccggtctgtactcactgtccagtgtcgtaaocgtcccttctagcag aacccagacctacatctgtaacgtcaaccataaaccatccaacacaaaggtggataagaaggttgaaccaaag agctgtgataagacacatacatgccctccttgtcctgcaccagagctcctcggaggtccatctgtgttcctgtttcccccca aacccaaggacactcttatgatctctcgtactccagaggtcacctgtgttgttgtcgacgtgagccatgaagatcccgagg tcaactggtacgtggatggagtcgaggttcacaatgccaagaccaagcccagggaggagcaatataattctaca tatcgggtagtgagcgttctgaccgtgctccac'caagattggctcaatggaaaagagéacaagtgcaaggtgtccaacaa ggctcttcccgctcccattgagaaaactatctccaaagccaaggggcagccacgggaaccccaggtgtatacattgccc ccatctagagacgagctgaccaagaaccaggtgagtctcacttgtctggtcaaggggttttacccttctgacattgctgtag agtgggagtctaacggacagccagaaaacaactacaagacaactcccccagtgctggacagcgacgggaocttcttcc t3:CCttagCCC huCD37—51 E; 3:5 I httggCCCtggafiCWcavu‘tCC210gC2121320bgécptgiatc‘gdtgggCtaCa‘tCCattdCagto0th23C. caattaCabCCCtagCCth2tggchgiatCtCt2itt21CCa2gataowtts:attaaCCagtttttCCta"thC‘taattCC9C; 3f gaCtg‘CctCtoacacama21CttactattvcgCCCUthCtdCtan.2CooattCCtwtttfitdtathggt'10222 C21CCC’Cg3‘1C21tgtctcagCchctctaCCCagC,gCCu2ttcagttttCCCCttggC3CCdagttctaaatCCaCattacwt ggaacagC‘thCCtgggathtctC1gttaaagattatttCCCtgagCCtgtgacagtgagctggaatagszggagcattgaact; tcaggtgtgcacaCttttCCCgctgtgttgcagtCCtCCO‘gtCtgtathqctgtccaototcUt‘)accgtcwttctaficag CttgggaaCCcagaCCmca[CtgtmtcgtcaaCcataaaCCatCCaacaCa'ta‘wtgattagaagflttmeC212mg agctgtgataagaCacataCzathCCtCCttgtCCtttcaC‘mgivtCCtCW-raggtCCat "tottmtvtttCCCCcca aggacacEcttatgatctctcgtactCcagaggtczwtgtgttott CgaCng‘ioCCaI‘Fa1g2ttCCCg23gg 3 ttaaattcaactggtacgtggatggagtcbItogttmcaatt.23a3g21-3C412ngccagggavoagCaatCtdattCtaCa 3 tatcgggtagtwavcgttctgdchthtCCaCC'tauttOgCthatggaaaw233132123.10tgc22309tC‘tCC23'x33121 = ggctcttCCCCgCtCCC‘a‘ttg.agaaddttatCtCCaa'thcaaggggCcagCCaC;’gfidLCLCEi};jtgtatamtthCC 3 CcatCtaC'tga2,0210:1033.caagaaCCaggtgagtCtCadtfitCtgg2C221gg0Ctttt21CCC:tts:tgaCattoctotag 2 agtaggagtctaacggaeagCcag21213123C21aCt2tCa31g2‘3CaaCtC‘C-CCC.gtgutggacagngngbgCttcttCC C tCt21CtCCaagjtgactstavacaagtctagatgngagC:a2:maaaCC‘ttttctcctgctcaataatm:atgaggCtCttha . 3 ‘ act "3%...EEEQ 11} N31):t."‘3!3m C C2121tC23C 21312233203: ‘taaatCaCtCtCC‘Cflax- C "huCD37—56 3 aagC‘tthCC‘CCt,gggtggagcthtttatCCtgttCCtCchCC‘aCCgmaCCgCCCtCCaCtCCC100twafict 3 3 3 aagcgggCCaggattggtaadm/CttCCCdCtCtCtoaUtcttacttgt1CCUt31tCtggammgtatcacatct 3 ggCttcgcctggmttgcattCUCC‘mtttCCCCgo023121;;quCttoafitgfiatgttggtCtCttthttCttCt 3 _ ngtacca 3 21CtacaaC:ccttCCCtgaagagtC9.21gtCtCaat‘taCCgCCamCttCcaagaaCC‘aattcttttt.gCagCttaattcagtg t :1CQETCCULC.021‘thng-tacttactactpcgcccgC,ggL'E3-i€.21di‘ igtttggtgcctggttcb7CCta.CtggggCC-2tggC—r Ua.CCCtggthCCgttgCtCC.gCCtCCamaaggm‘C‘atCattttCCCCttCCCtCC2121CttC232‘231tCcacaagC tCCaavmtctgca.gggatoCCtCott2t21gfittatttCCC‘tgaCtgtUaCacwagtggadtdfiCCgaCCattCaC tthcgtotgcaCaCthCCovtotgttCC213‘CCCtCngtCtgtz1thatothaotCtCCtadCgtCCCttCtzth223g Cttgggaa»;‘CC213123103:moatctgtaantC-aaccat21321ccathaaCacaztagotgoat’taoaaggttva 21CC2321220 agthtgdtawacaC‘ataCztthCCtCCHf‘TCQT‘T"aCCuC,"'iCCthUdg C2 otgttCCtot‘tCCCCMa asCCCadggaCaCtCttCtgatctctCotCCtCCagaggtCax.CtCtUttqttgtC322Cgtg C"-W2312102131901»agg ttaaattcaaCtgntaCGtggmggaCCCCaUCttcacmtgcm022C021m.CCCoggL,magcadt1tC2222C.2tCa tatcgggtagtoaocgttCtoaCCgtUthcaCcaagattv0CtwatggaaaagagtacQagtfiCdagCtgtCCaacaa 2 gCtCtthchtCCC2ttt-1omaact'ttCthttawccamggwammwg1CCCC2‘281‘tot‘1taCrttth" 93CtCtagmaCtzagCt1221mm210222103.aggtvagtctcm‘t‘rtctvgt21215.23;>ttLC1CCCttCtgaCCttvctgttg:94 3 aggtC'vagtCt212tC0gCtC1w3.2.3232212121C213taCaaga.Ca21CtCCCCC2tgthtggaCagCganggagCttCtth 3 tcta‘thaaCtthtCtnthaC‘aaCtC3.102atagcs10‘ca2tUgaattcyttttttcctgctCagtaatgcatgaggCtctgca 5 3 " ID 330360) 3 Cgtathggtt'tnCC132222121{nggitCCCttdz- CCangtoaCtC ______________________________ huCD37—57 CCCaCcatgggCtggwgC g :atcattCtgrttCtootgC acaccadCthCttcaC2339:0212TtCCCastj cagggagCLigCC-C‘Cfiga'tCC-toaadCcattCtCagtcaCt23Cgtctg.acatvtathtCa’fCWCtaigcattacctc‘ aggcttc37:0ttggC2 ttvodtmeathCCCCgaaaACotCt;:10tggatgvggmC~22ttCtgt2tC2thggC21gta 1 C2132EtattCaCCCtCCtt:32:atCtdggatatCattCfiCaCCtCatacafi1.0213213at2WC1YLC‘C"‘§.,CE§ZQC£ECTV" 3 gtcaCCCgcchagaC21C30CaaCCtattdttUthtwggflfiCtacggaCCtgfichatayttCOCCtdtttzwrszC1 3 gr:g23C2CLCCYLUaCCthuchchCCCC213212321ggCCC231C210ttttCCCCtthCtCCmf‘ttctuaatCCdeg ngtggaaCarrCt'iCdCtgggatCCCthttaaaoattatttCCCtUagCCCgtCaCaCt"Ct'roaatagcggagCat tgacttmggtgt""23C23CttttCCCUC{Giotto&OtkCECCQgTLfgtaCTKaLTgtC2tgtfitCm312tCCrrtwcttCta gcagcttoomaCCCagaCCtaatCttftaaCCtCaaC312323233.CatCCaaCaCaaaqgtggatadzaaftgttgaaCC 3122210310C10tgfiaagacatcatacatguCCtCCttUtCCthCthamCCtCCtcggaggts,catCtgtgtthtgtttCCC CCC323aCCCa'tngdCtCttawatCtCthtaCa0agatcaCCtgCattwttgtCU'tcvtgagC23tg2331g2‘3tCCC gaggttaaattCaathtngggdtggwtm UgttcaCaatgccaagaCCaaOCCCdgg‘21g31agC21-21t2tt2321tt tta‘tcgggt21;;thCgt:gttCtCachtCC‘t31CC121g31ttvtiCtCa2atggaaaagagtacaaotttcaaggtvtCC aacaaggctcttccCcg‘CCitCCCttmaaaaactatCtCcaaaUCCaaggcagCCanwaaCCcaggtetatacat catCtagggacga C ggggttttaCCcttCtgawttg ‘ ' ‘ ‘ 2 ' " ' ' " " 32233 <‘"‘221C\’2.2*agc 4 4 4.4."\\\L"..........,,_.w~aaaL""A. Caiaatgcafgawc"LLLLLLLLLLL‘‘L w________ L"\"FL‘LLLWLLLLW" d E chCD373 Od&EECELLfiLCLHé3thLLoaLiLagtLLtgggLfigctotthgIOthtaC'xgatg"*agaigtLaLa‘iLLag LagiLmLagL.CECLL‘LE’ELtgiaioigtgggagaaadgtcaooatcaoatgtLgagoaagigagaaLaitLgCa gtaatttag0atggtatoagcagaaamgggaaaatotoCtCagCEcctggmaaigflgcaacaaaofiagoagatggtgi gCLaLoaagCfiLagtvgcagtfi‘O‘atcaggcaoaoagtafiocottcaagatcaaoagcctgcagtczgaagatmggga CfiattactgtoaacattaitggggtaotacgiggaogficthgaggoaccaagotggaaatcaaaogtacQfitggow cazccatLtgtofiLatLt gaocaOECaaatLtggaaLfgLoiatgitgwigCotontoaamaLfiota tCLCagagaggoLaaagtaCaoioLaaggtogataachLoC’iLoaatgggiaaotoccaggagagtgto3Cagagc arigéLaocaaggaagoaoctaLCOLL?‘agoagoaocotgacgotgam433LgLagdLiaC"agdaawcaaaot oiaC0LLthoaaotLaLoLanagggoctgagotogcocgtoaoaaagagcitLaacaggggagagtgfiag3 v"n"... .LLLLLLLLLLLLLL" """LLLL‘LLL—Mg"4.(son)3359.162) mhuCD37-3 aa’tLgCLaLLatgth . LgLaLLaLttott:CtgigchaoagocaccggiLtiLauLigatamLaaatoaL (1.0 and 1.1) i ECLiachLttCLagtttgtaagigtjggigmgcgtaacaatoaoLtomgdLoiagiaaaacaiccgoagta 231i.thLaiggtacoaaCaaaagoLaggiaavts:aLLtaaacthtc:ItomtwfigotuowCLtcvotgatvgtUtgo oiioaogatiLiLiggficaggttongLoamfioatiadgaicaaLtLaLtLoaaCLaCaagaitingiacama LtgtoficatomC4ogcoatotgatgigcagttgaaatctggaaotgogc:"LCgtgLLLgLigaaiaaLttCtaICLod gagaggoCaaavtaoagtggqawtggataaogcoctcoaatcgggtaaLE'LCLL4aggg.smgtglLaciigagoaggao L deaagLacamaLLtaoaOCLtLagCgem:oCtgachtgagcaaagcaqaL wgagaaaczxsaaagtotacgc L K : 3 otfiomagime canggLotGaCLtgoC-CgiLacaadgaCLtiLaai'gggagagtgttag (SEQ ID C NO: 163) \ chCD3712 \ gamogLCaCLai0ggflggtofigtataatCLtgttLtivvEggLL4chLgLmagchmgataotgutafigiactoact \ "Mann"...m... \ CGgtoaooagcoagtCtggCagiothLtggPocagcgtgocaocaCicctgcogggoLtoaLjttoCGIgaooaCta 3 gctcfiafiootatciotaotggtfio:aLagaeLocCaggasacoCLoiaLg:gtCigatcaagt'(LochtccaaCCrcgo \ CagoggcortwoootagafioLLiovtiLCoLLwCooaaotoamoao‘mgaaLa{LLaLLLLgitggggaagagga _.... taCLgCLaLfiaLLtaItgtLaacaotcfigggacattLLKaLaLLm0Gaagamaaoaaagotogaaatt'aagcgtzwg Wfluflfl."flu—"y,"5...... gtggLWLdL:tctgtcfioatcttcocCLC:1thgatgaUCaatLgaaatN0gaaothCtotvfificivodgdgdai mtcocagagaggLfaaagtaCagtgoa.aag.g0aiaaLCLLLiLLaatLLgUEaaL{L‘ooao0agagtha caoagoaggaLagCaagg:agCaLLtoLagLotoaocavmCoigac(5cigagLaz=a0Ca<>aLtacgavaaaLa L'aaagtotaorrLLtoLgaagtaCCLatcagggoCdgagotogcocgt'tascaaagzthtLaamggaqagangfia I Nmmmmw" -~—»~»--—»« ~- —-— chCD37—3 8 LaLC'ttghLLiggtLLtg afLatCLLgtttLtLgtggoLaoamacéggtgficaitchagafimgdgdoof LaatLaLoaUCEafiatgtLogLagerCLCggCoagaaaatfiaLmioaatgtzagC2L1:gthctotgacfiaoaf ...... _;cattggtatoaaLagJAgLL,ggtaco:chC"aanogfiggaflmogaodwfiw ggLLthggaotLCLtCa4 LCigjL‘LCanggaootgnmooggoaLagtiatLaLtgchatatLCLCtaLfivm:igaagaUerLLaLmnta itgtcaaoaatggamotaaoufiCCocoaLCitggthngaaLadagLtgflagangogaLLUtfigLWCaLLaL L otgtottLElK/HLLLLLKatLtgp'Hg‘hOk/imga"(1E LaaLLGLLLLtottotthCtgotgaataaaofictatLLoa \ gagaggoasaviacafitgmawtggataaLgCLCCcoaa‘ngggiaCocaggagagtgtcawoaaoaqgao 3 .LgceLaggaL30LaCLtaLavLLtageagoaCCC.gacwctgagL(1:130agauaogagameaaagtctacgo otgLg3.agtLaLCCaimgggoowc,agotcgCLgodagaagathiLaacaggggagagtgtiag (SEQ T53 M3365) .34._V_______.,...........u..auu"an"............. huLDfl38 C70tLogLLaoLaiOggatggtLLtgLafiafttgmflwtwocacgLCBCtggficaLLo‘macmotgtcaca .agtC‘LCLagLLLLaLtg[LigLi‘tCCLLoootfiagooggtvaLoatgaLgctotgCcagficotoogtgacatamtgo Ci‘LaaaaaoatggmotaCoaLaottcaaaaofigcat'avgogfioctocoa ‘ attogtatoagoaaaaaoccvgt'o 0215.333:""5bgifliflgbtvtoobfiticscatacagtotngcattagtELLatogaacdggaga’rgcagccaoCmLiaCtgt aowvavaiaaam0&3.ngggLot-cacoa‘mfsw ""17".L" .w m"L""LL‘L‘LLL‘LL. -69.. icttcatcttCCCOCcatCtgatgaoC.agtwaaatctggaaC.EgCCtCtJttgtgLCEgCEgaataaCCtC.taECCCagagaogccaaagtaCagtggaarrgttwatcmCg1.CCECcaachggtadCtCCCaggagagtomEEC-3oaacavgacaficaaggz(ECCagCaCCtaCCgCCtCagcagCaCCCtCaCCctfiagaaaocawCECECCCwaaaCaCaaaotCEaCCCthCgaagECaCCatCagggcctgagCtCUCCCgECaCaCCagagCthaaCanggdrragwttag (SEQ ED0 CCCCCCCCCCC ........ CCCCCCCCCCCCCCCCCCC CCC. CCCCCCCCCCCCCCCCCC CCC__C._.....C "(11:thaCCatzqgttogtCatocatwtCtgttCCCCqer’CaC0C1ggagtauiawtwCECE’EO‘ECCELEEC tCCthtdCtatgtCth.CagcCCCEggagdgCgtgtgamatgacttuELEgLECELCCaa‘VtgtgaCatacar qcattggtatCagcaaaagCCtggCCCa‘CCCCCtaaaaggtggatctacgataCttCtaatcthCatangtgEUCCC0C aaggttCtCCGggagtgg"agtvgcaCCCOfiatagtctC'tccatcavttcaatggaagcagaggatgcagcaaCCtdttC attotCauCagtogthgtttaatCCCCCtaCtttthICItggtaCaaag-tvgaoaEtaagcgtacge’tggctficacmt ctgtCEtCatctEC.CCUCCatCtgatgagcagttgaaatctooaactscCCctfittgtgtOCCEgCtgaataaCtECtatCCCC21 1 3" "109:1:GM 52':Q {If}0 ('3 '99E"C.)ionIn,200 330HQa»).0‘"(IQ$1)(7%33H"E3321:E35 (-£9 {:3O 0"? {'3C ('215-4» ('130 )W(38OCE‘TJfl3) 311)(D ,4.(3 (.3 (2 a: 0‘53 (3:; 9.2(1'2"‘3:'3».(—1- If;P4 ('2m f") {3) (TC? ($2)?..... agCaaggaCagCaC.CtaCagCCtCCgCagCCECCCtg1.OCtoagCaaagCagaC‘CCECvagaaCCmagic50381894 Cg CthgaagtcaCCCCtCCEggchtgaqctchCCCCgtCCC2aangCttCaCCCgggaoagtgttavg‘EEQ EDC huLD3/5E" gaa‘ttchC.caCatgggamgaCctgtattattCtottCtggtthtaCtht'ECtgCgt CCaaaaacCag113301CCCgtcgttggatttaEgatCtCCtCtam.CEOOCEECagCgEtCCtg 1 1 ttt‘Ctttotaatgg'ttngggCttcctdEaOCCttaCCataaOCECCCthoaaUcCwaawCCgCtacataCta 1 ctgccagcagtggagtgataaCC.CCCCCCCCtthggCaggngCCaaat‘ggaoatcaaacgtangtggCthCCC 1 atctgtCtticatCE‘ECchCCCtCtgatgagcagttgaaatCtggaacEgCCtCtoEtOEgtoCctgctgaataacttctaCC 1310aGagchaaagtaCaCztgfiaaggtgataacocCctCC'thWstdaCtCCCaggagaotgtcaCCC-agt.gga cattcaaggacaUCdCCtaCCGCC[CagC'EgCa.CCtganCtgdt—CcaaetvcagactaCgagaaacacaaagtctacg g CCthgaaotCCathggg-CtgagctcgCCCgtcacaaagagc :1acaggggagagtgttag (SEQ EEE N0E201 5135225} Also pmvECECCE is a poEymacEeoEECEC having at ECast about 95%,, at ECaSt about. 96%. at. 11:21:11; about 97%, at 583551: about 98%, m at ECast about 99% C ty to SEQ ED NOS:E‘2E—E7€), E83. or E83.

Thus, in 12611211311 em‘bvdiments, the poEynucECCtiCEC comprises (a) a. poEynuCECC‘CiCEC E1avi1‘1g at ECasE about 95/6 sequence ECECEEEEEV t0 ‘1)"Q ED N05: 121‘ ~ E 35 E524. E, or 182., and/01" (E3) 21 pCEynuCECCtECEC having at 11321211: Q5/CSCC1CCCCC Edcnttty to SEQ ED NOS: E36 ESE E62 E76, 0': E8E. E11 Cartam embodnnents. the .. 7g} - polynucieotide comprises (a) a cleotide having the nucieic acid sequence of SEQ II.) NQs: 121— 135, 152-161 or 132; and/or (b) a polynucleotide having the nucleic acid sequence of SFQ II.) NOS: 136»- 151,162—170, or .183.

In same embodiments, the pclyiiucleetide encodes the iight chain encoded by the recnmbinant d DNA phuCD37—3LC (ATCC Deposit Designation PTA-10722, deposited with the ATCC on March 18, 2010) or a light chain that is at least about 85%, at least about 90%, at least about 95%, or at least about 99% to the light chain encoded by phuCD37-3LC (PTA-10722). In some embodiments, the polynucleotide encodes the heavy chain encoded by the recombinant d DNA phuCD37-3HCV.1.0 (ATCC Deposit Designation PTA-10723, deposited with the ATCC on March 18, 2010) or a heavy chain that is at least about 85%, at lesat about 90%, at least about 95%, or at least about 99% identical to the heavy chain encoded by phuCD37—3HCV.1.0 (PTA—10723). In n embodiments the polynucleotide is the recombinant plasmid DNA phuCD37-3LC 0722) or the recombinant plasmid phuCD37- 3HCv.1.0 (PTA—10723).

In n ments the polynucleotides comprise the coding sequence for the mature polypeptide fused in the same reading frame to a polynucleotide which aids, for example, in expression and ion of a polypeptide from a host cell (e.g. a leader sequence which functions as a secretory The polypeptide having a leader sequence for controlling transport of a polypeptide from the cell). sequence is a preprotein and can have the leader sequence cleaved by the host cell to form the mature form of the polypeptide. The polynucleotides can also encode for a proprotein which is the mature protein plus additional 5' amino acid es. A mature protein having a prosequence is a proprotein and is an inactive form of the protein. Once the prosequence is cleaved an active mature protein remains.

In certain embodiments the polynucleotides comprise the coding sequence for the mature polypeptide fused in the same reading frame to a marker sequence that allows, for example, for purification of the encoded polypeptide. For example, the marker sequence can be a hexa—histidine tag supplied by a pQE-9 vector to provide for purification of the mature polypeptide fused to the marker in the case of a bacterial host, or the marker sequence can be a hemagglutinin (HA) tag derived from the influenza hemagglutinin protein when a mammalian host (e.g. COS-7 cells) is used.

The present ion further s to variants of the hereinabove described polynucleotides encoding, for e, nts, analogs, and derivatives.

The polynucleotide variants can contain alterations in the coding regions, non-coding regions, or both. In some embodiments the polynucleotide variants contain alterations which produce silent substitutions, ons, or deletions, but do not alter the properties or activities of the encoded polypeptide. In some embodiments, nucleotide variants are ed by silent substitutions due to the degeneracy of the genetic code. Polynucleotide variants can be produced for a variety of reasons, e.g., to -7}- optimize cedon expression for a uiar host ifchange sedans in the human mRNA to those preferred by a bacterial host such as E.. call). {00231} Vectors and cells sing the polynucleetides described herein are also provided.

IV. Metheds 9? use and plmmraeetxticai eompesitiens £00232] The CD37—binding agents (including antibodies, immunoconjugates, and polypeptides) of the invention are useful in a y of applications including, but not limited to, therapeutic treatment s, such as the ent of cancer, such as B—cell malignancies, autoimmune diseases, and inflammatory diseases. In certain embodiments, the agents are useful for depleting B-cells. In certain embodiments, the agents are useful for ing autoreactive B—cells. In certain embodiments, the agents are useful for depleting peripheral B-cells. In certain ments, the agents are useful for preventing inappropriate T—cell stimulation. The T-cell stimulation can be in connection with a B-cell y. The methods of use can be in vitro, ex vivo, or in vivo methods. In certain embodiments, the CD37-binding agent or antibody or immunoconjugate, or ptide is an antagonist of the human CD37 to which it binds.

] In one aspect, anti—CD37 antibodies and immunoconjugates of the invention are useful for detecting the presence of CD37 in a biological sample. The term "detecting" as used herein encompasses quantitative or ative detection. In certain embodiments, a biological sample comprises a cell or tissue. In certain embodiments, such tissues include tissues that express CD37 at higher levels relative to other tissues, for example, B—cells and/or B—cell associated tissues.

] In one aspect, the invention es a method of detecting the presence of CD37 in a biological sample. In certain embodiments, the method comprises contacting the biological sample with an anti-CD37 antibodyunder conditioes permissive for binding of the anti-CD37 antibody to CD37, and detecting whether a complex is formed between the anti—CD37 antibody and CD3 7.

In one aspect, the invention provides a method of diagnosing a disorder ated with increased expression of CD37. In certain embodiments, the method comprises contacting a test cell with an anti-CD37 antibody; determining the level of expression (either quantitatively or qualitatively) of CD37 by the test cell by detecting binding of the anti-CD37 antibody to CD37; and comparing the level of expression of CD37 by the test cell with the level of expression of CD37 by a control cell (e.g., a normal cell of the same tissue origin as the test cell or a cell that expresses CD37 at levels comparable to such a normal cell), wherein a higher level of expression of CD37 by the test cell as compared to the control cell indicates the presence of a disorder associated with increased sion of CD37. In certain embodiments, the test cell is obtained from an individual suspected of having an autoimmune disorder or atory er. In some embodiments, the disorder is associated with increased expression of CD37. In some embodiments, the disorder is ated with increased number of B—cells. in some ments, the disorder is associated with increased activity of B—cells- In certain embodiments, a method of diagnosis or detection, such as those described above, comprises detecting binding of an anti—CD37 antibody to CD37 expressed on the surface of a cell or in a membrane preparation obtained from a cell expressing CD37 on its surface. In certain embodiments, the method comprises contacting a cell with an anti-CD37 antibody under conditions permissive for g of the antiutZTDBTf antibody to {31337, and detecting whether a complex is formed between the anti—CD37 dy and CD37 on the cell surface. An exeniplaiy assay for detecting binding of an anti—CD}? antibody to (112337 expressed on the e ot‘a cell is a " assay. {68237} Certain other methods can he used to detect binding of a.nti~€3i>f57 antibodies to CD37. Such methods include, but are not d to, antigen~binding assays that are well known in the art, such as western blots, i‘adioimmunoassays, ELlSA (enzyme linked inimunosorbent assay), "sandwich" noassays, imniunoprecipitation assays, fluorescent iininnnoassays, protein A immunoassays, and inin‘iunohistocheinistry (EEC). {80238} in certain embodiments, anti—CD37 antibodies are labeled. Labels include, but are not limited to, labels or moieties that are detected directly (such as fluorescent, chromophodc, electron-dense, chemiluminescent, and radioactive labels), as well as moieties, such as enzymes or ligands, that are detected indirectly, eg, h an enzymatic on or molecular interaction. {00239} In certain embodiments, anti—CD37 antibodies are immobilized. on an insoluble matrix. immobilization entails separating the anti-Cl337 antibody from any CD317 that remains free in solution.

This conventionally is accomplished by either insoltibilizing the anti-CD37 antibody before the assay procedure, as by adsorption to a waterninsoluble matrix or surface (Bennich et at, US. Pat. No 3,72 ,760), or by covalent coupling (for e, using glutaraldehyde crossulinlting), or by hilizing the anti~CDL¥ 7' antibody after formation of a complex between the anti-{7937 antibody and CD37, e..,g by imntnnoprecipitation.

: Any of the above embodiments of diagnosis or detection can be carried out using an imntunoconiugate ofthe invention in place of or in addition to an D37 antibody. in certain embodiments, the disease treated with the C3D37~hinding agent is an autoimmune or atory disease. In n embodiments, the autoimmune or inflammatory disease is selected from the group consisting of psoriasis, itis, systemic derma and sis, ses associated with inflammatory bowel disease, Crohn's disease, ulcerative colitis, respiratory distress syndrome, adult atory distress syndrome (ARDS), dermatitis, meningitis, encephalitis, uveitis, colitis, glomerulonephritis, allergic conditions, eczema, asthma, conditions involving infiltration of T cells and chronic inflammatory responses, atherosclerosis, leukocyte adhesion deficiency, rheumatoid tis, systemic lupus erythematosus (SLE), diabetes mellitus, multiple sclerosis, Reynaud’s syndrome, autoimmune thyroiditis, allergic alomyelitis, Sjorgen's syndrome, juvenile onset diabetes, immune ed by cytokiiics and "illymphocytes, responses associated with acute and delayed hypersensitivity tuberculosis, sarcoidosis, poiymyositis, otnaiosis, vasculitis, pernicious anemia (Addison’s disease), es involving leukocyte dispedesis eentt’ai nervous system (CNS) inflammatory disorder, multiple organ injury syndrome, hetnolytie anemia, myasthenia gravis, antigetnantihody compEex mediated diseases, anti~glomerular nt membrane disease, antiphosphoiipid syndrome, aiiergic neuritis, Graves disease, tEaton myasthenic me, petnphigoid buElous igus, autoimmune polyendocrinopathies, Reiter’s disease, stiff—man syndrome, Behcet disease, giant cell arteritis, immune complex nephritis, lgA nephropsthy, igM polyneuropathies, thic thromboeytopenic a (Ill?) and autoimmune throtnhocy‘topenia. {@242} in some embodiments, the autoimmune or inflammatory disease is seieeted from the group consisting of: RA, inpus, immune thromhocytopenic purpurn, pure red ceiE aplasis, autoimmune anemia, coid agglutinin disease, type B syndrome of severe insulin resistance, mixed cryoglobuEinermis, ntyasthenia gravis, Wegener's granulomntosis, microscopic polyangiitis (MFA) refractory pemphigus vulget'is, dermatomyositis, Sjogren’s syndrome, active type—ii mixed cryogiobuiinetnia, petnphigus Viligai‘is, autoimmune neuropatliy, paraneopiastic opsocionusarnyoclonus syndrome, and, relapsing ing ntuitipies osi s (ERA/ESE {(33243} int:ertain embodiments the autoimmune disease or inflammatory diseaseiscltarecterized by CD37 xptess1n- eelis to which the nding agent (eg.,antibody) binds, {.6244} Ehe present invention provides Eor methods of ng autoimmune and intiannnatory diseass tompiising adtninistetingaatherapeutically effective amount of a CD3'7—bihding agent to a subject (tag, a suhject in need oE’treatment), in certain embodiments, the subject is a human. {@245} The present ion further provides methods for depleting B-cells, e,g., autorcuctive B» cells, .sittgth antibodies or other agent: described herein. En CRttain ments, the method of depleting B—ceiEs comprises contacting a B~ceil with a CD37~hinding agent. (eta, antibody) in vitro. For example, a eeEl Eine that expresses CD37 is cultured in medium to which is added the antibody or other agent to deplete the cells. in some embodiments, the ceEEs are isoiated from a patient sanmie such as, for exatnpie, a tissue biopsy, i effusion, or blood sample and cultured in medium to which is added an CD3 7~hinding agent to deplete the. ceils. {@8246} in some etrthoditnents, the method of depleting B~eells, eg. autoretetive ls comprises ting the cells with the C3E337~hinding agent (cg, antibody) in viva, in certain embodiments, ting a red with 3 inding agent is undertaken in an animal model For example, CD37- binding agents can be administered to xenografts expressing one or more CD375 that have been grown in immunocompromised mice (e.g. NOD/SCID mice). In some embodiments, cells are isolated from a patient sample such as, for example, a tissue biopsy, pleural on, or blood sample and injected into immunocompromised mice that are then administered a CD37—binding agent to deplete B-cells. In some embodiments, the CD3 7-binding agent is administered at the same time or shortly after introduction of cells into the animal. In further examples, CD37 binding agents can be administered in vivo to mice expressing one or more CD37 antigens. In some embodiments, these mice can be engineered to express human CD37 in addition to, or instead of, murine CD37. In some embodiments, these mice are disease models, e.g. models for autoimmune disease. In some embodiments, administering a CD37 binding agent es B—cells in vivo. In some embodiments, a CD37 g agent prevents T—cell stimulation. In some embodiments, administering a CD37 binding agent prevents or alleviates an mune disease. [00247; in certain ments, the B~celis overexpress CD37, in ether embodiments, the B-ceils do not overexpress CD37. In some embodiments, the B—cells are not cancer cells. in some embodiments, the IRS—cells are not turner ceiis In some ments, the B-cells are not cancerous cells.

The present invention further provides pharmaceutical compositions comprising one or more of the CD37-binding agents described herein. In n ments, the pharmaceutical compositions further comprise a pharmaceutically acceptable vehicle. These pharmaceutical compositions find use in treating autoimmune and inflammatory disease in human patients.

In certain ments, formulations are prepared for storage and use by combining a purified antibody or agent of the present invention with a pharmaceutically acceptable vehicle (e.g. carrier, excipient) (Remington, The Science and Practice of Pharmacy 20th Edition Mack Publishing, 2000). Suitable pharmaceutically acceptable vehicles include, but are not limited to, nontoxic buffers such as ate, citrate, and other organic acids; salts such as sodium de; antioxidants including ascorbic acid and methionine; preservatives (e.g. octadecyldimethylbenzyl ammonium chloride; thonium de; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight polypeptides (e.g. less than about 10 amino acid residues); ns such as serum albumin, gelatin, or immunoglobulins; hilic polymers such as polyvinylpyrrolidone', amino acids such as glycine, ine, asparagine, histidine, arginine, or lysine; carboEydrates such as cchandes, disaccharides, glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or ol; salt-forming counter-ions such as sodium; metal complexes (e.g. Zn-protein complexes); and non—ionic surfactants such as TWEEN or polyethylene glycol (PEG).

The pharmaceutical itions of the present invention can be administered in any number of ways for either local or systemic treatment. Administration can be topical (such as to mucous membranes including vaginal and rectal delivery) such as transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders; ary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal); oral; or parenteral including enous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial (e.g., hecal or intraventricular) administration, -75..

An antibody or immunoconjugate of the invention can be combined in a pharmaceutical combination formulation, or dosing regimen as combination therapy, with a second compound having anti-autoimmune or inflammatory ties. The second compound of the pharmaceutical combination formulation or dosing regimen can have complementary activities to CD37—binding agent of the combination such that they do not adversely affect each other. Pharmaceutical compositions comprising the CD37—binding agent and the second agent are also ed. For example, CD3 ing agents can be administered in combination with CD20-binding agents, such as Rituximab. In other embodiments, CD37—binding agents can be administered in combination with salicylate‘, nonsteroidal anti-inflammatory drugs such as indomethacin, phenylbutazone, phenylacetic acid derivatives (e.g., ibuprofen and fenoprofen), naphthalene acetic acids (naproxen), pyrrolealkanoic acid in), indoleacetic acids (sulindac), halogenated anthranilic acid (meclofenamate sodium), piroxicam, zomepirac and diflunisal; antimalarials such as chloroquine; gold salts; penicillamine; or suppressive agents such as methotrexate or corticosteroids. In some embodiments, the CD37-binding agent is administered in combination with a second therapeutic selected from the group ting of methotrexate, an anti-CD20 therapeutic, an anti—IL—6 receptor therapeutic, an L-12/23p40 therapeutic, a chemotherapeutic, an immunosuppressant, an anti-interferon beta-1a therapeutic, glatirareer e, an anti—a4-integrin therapeutic, fingolimod, an anti-BLys eutic, CTLA-Fc, or an anti-TNF therapeutic. In some ments, the CD37—binding agent is administered in ation with a second therapeutic that is an antibody directed against an antigen selected from a group consisting of CD3, CD14, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD36, CD38, CD40, CD44, CD52, CD55, CD59, CD56, CD70, CD79, CD80, CD103, CD134, CD137, CD138, and CD152. In some embodiments, the CD37—binding agent is administered in combination with a second peutic that is an antibody directed against a target selected from the group consisting of IL-2, IL—6, IL-12, IL-23, IL-12/23 p40, IL-17, IFNy, TNFOL, IFNOL, IL—15, IL-21, IL—la, IL-lb, IL—18, IL—8, IL-4, GM—CSF, IL—3, and IL—5. In some embodiments, the CD3 7-binding agents are administered in combination with methotrexate.

For the treatment of the disease, the appropriate dosage of an antibody or agent of the present ion depends on the type of e to be d, the severity and course of the disease, the responsiveness of the disease, Whether the antibody or agent is administered for therapeutic or preventative purposes, previous therapy, patient’s clinical history, and so on all at the tion of the ng physician. The antibody or agent can be administered one time or over a series of treatments lasting from several days to several months, or until a cure is affected or a diminution of the disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient and will vary depending on the relative potency of an individual antibody or agent.

The administering physician can easily determine optimum dosages, dosing methodologies and repetition rates. In certain embodiments, dosage is from 0.01 pg to 100 mg per kg of body weight, and can be given -76— once or more daily, weekly, y or yearly, in certain embodiments, the antibody or ether CD3?— binding agent is given once every two weeks or once every three weeks. in certain embodiments, the dosage of the antibody or other CD3 ibinding agent is from about 0.1 mg tc abcnt 20 mg per kg of body weight. The treating physician can estimate repetiticn rates for dosing based on measured nce times and concentrations of the drag in bodily fluids or s. {9:325:23} The combination therapy can provide "synergy" and prove "synergistic", ie. the effect ed when the active ingredients used together is greater than the sum of the effects that results from using the compounds separately. A synergistic effect can be attained when the active ingredients are: (l) mulatcd and administered er red simultaneousiy in a combined, unit desage formulation; (2') delivered by alternation or in parallei as separate formulations; or (3) by some other regimen. When delivered in alternation therapy, a synergistic effect can be ed when the cempcnnds are administered cr delivered sequentiaiiy, eug by different injections in separate syringes. in general, during aiternation therapy, an effective desage of each active ingredient is administered sequentially, i..e serially, whereas in combination y, effective dosages of two or more active ingredients are administered together, VI. Kits comprising CD37-binding agents The present invention provides kits that comprise the antibodies, immunoconjugates or other agents described herein and that can be used to perform the methods described herein. In certain embodiments, a kit comprises at least one purified antibody against CD37 in one or more containers. In some embodiments, the kits n all of the components necessary and/or sufficient to perform a detection assay, including all controls, directions for performing assays, and any necessary software for analysis and presentation of results. A label or indicator describing, or a set of instructions for use of, kit components in a ligand detection method of the t invention, can also be included. The instructions thereof. One may be associated with a package insert and/or the packaging of the kit or the components skilled in the art will readily recognize that the disclosed antibodies, immunoconjugates or other agents of the present invention can be y incorporated into one of the established kit formats which are well known in the art. Such kits can also include, for example, other compounds and/or compositions, a device(s) for administering the compounds and/or compositions, and written instructions in a form ibed by a mental agency regulating the manufacture, use or sale of pharmaceuticals or biological ts.

Further provided are kits sing a CD37-binding agent (e.g., a CD37-binding antibody), as well as a second agent. In certain embodiments, the second agent is mab. In certain embodiments, the second agent is methotrexate.

* * * Embodiments of the present disciosure can be further defined by reference to the following non-limiting examples, which describe in detail preparation of certain antibodies of the t disclosure and methods for using dies of the present sure. It will be apparent to those skilled in the art that many modifications, both to materials and methods, can be practiced without departing from the scope of the present disclosure.

Examples It is understood that the examples and embodiments described herein are for illustz‘ative es only and that various modifications or changes light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application.

All publications, patents, patent applications, internet sites, and accession numbers/database cited herein are hereby incorporated sequences (including both polynucleotide and polypeptide sequences) by reference in their entirety for all purposes to the same extent as if each individual publication, patent, and individually patent ation, intemet site, or accession number/database sequence were specifically ted to be so incorporated by reference.

Example 1 CDstXUesswnInnomalhumanJlBMCs The CD37 antigen was reported to be expressed on B-cells from the pre—B stage to the eral mature B—cell stage, while being absent on B-cell progenitors and terminally differentiated plasma cells. (Link et al., 1987, J Pathol. 152:12-21). In addition, the CD37 antigen is only weakly expressed on T—cells, myeloid cells and granulocytes (Schwartz-Albiez et al. 1988, J. Immunol, 140(3)905—914).

The ability of antibodies ding certain CD37 antibodies and immunoconguates previously described in US. hed Application No. 256153, which is herein incorporated by reference in its entirety) to bind to normal human B—cells was measured using flow cytometry assays with fluorescently labeled antibodies. In addition, the cially available QuantiBRITE system from BD Biosciences was used to estimate antigen density based on the number of antibodies bound to the cells (ABC). The QuantiBRITE system from BD ences utilizes the following reagents: anti-CDZO-PE supplied at 100 ug/mL and QuantiBRITE PE supplied as lyophilized PE-labeled beads. In addition, the huCD37-3 antibody was labeled with PE to obtain an antibody—PE conjugate with an AbzPE ratio of approximately lzl.

Fresh buffy coats from healthy donors were obtained from Research Blood Components ton, MA, US) as a source of normal blood cells. Buffy coats were prepared by centrifugation of a unit of whole blood and collecting the interface between the plasma and the red blood cells. This unpurified buffy coat contains PBMCs, neutrophils, platelets, red blood cells, and plasma and was used mfg- for experiments on the same day it was drawn. Peripheral blood mononuclear cells ) were prepared from buffy coats by standard density gradient centrifugation using Ficoll-Paque as follows.

Blood was diluted 1:3 with 1x HBSS containing 5mM EDTA and up to 30 mL were added to a 50 mL conical tube. Ten mL of Ficoll-Paque (GE Healthcare) were slowly added to the bottom of each tube.

Samples were centrifuged at 500 x g with no brake at RT for 30 minutes to obtain a layer of PBMCs below the plasma and to remove red blood cells and most granulocytes. The PBMCs were transferred to new tubes and washed twice with 1x I-[BSS containing 5mM EDTA by centrifugation at 400 x g for s at RT. Staining buffer (1x HBSS, 1% BSA, 0.1% sodium azide) was then used to resuspend the PBMC pellets at 6.25 x 106 cells/mL. Eighty uL of cells were transferred to a round-bottom 96-well plate to achieve 5 x 105 cells/assay and 20 uL of human serum (Sigma H4522) were added to block Fc receptor- mediated binding and ted with cells on ice for 20 min in the dark. Fluorescently labeled antibodies obtained from Miltenyi were used to identify PBMC populations: anti-CD3-allophycocyanin (APC) was used to identify T-cells, Dl9-APC for B—cells, anti—CD56-APC for natural killer (NK) cells and anti-CD14—APC for monocytes.

Cells were co-stained for CD37 expression using 20 uL of huCD37—3-PE for a final concentration of approximately 10 ug/mL. Likewise, cells were co-stained for CD20 expression using 20 "L of anti-CD20—PE. As a control a nding PE-labeled hngGl isotype control antibody was used at ug/mL. ng was d out for 1 hour on ice in the dark. Samples were washed twice with staining buffer and fixed in 200 uL of 1% formaldehyde in 1x PBS. Samples were stored at 4°C in the dark until ition, which was performed within 4 days of sample preparation.

A fresh tube of QuantiBRITE beads was reconstituted in the supplied tube with 0.5 mL of staining buffer just prior to sample acquisition. Samples were acquired on a FACSCalibur flow cytometer (BD Biosciences). Compensation controls were run with each assay to select appropriate instrument settings and at least 10,000 events were ted for each sample. ment settings for fluorescence and sation were kept the same for both cell sample and bead sample ition to allow for an accurate comparison. CellQuest (version 5.2.1, BD Biosciences) was used for acquisition control and analysis.

The QuantiBRITE analysis utilizes on a bead standard with 4 bead populations conjugated with a known number of PE molecules. For data analysis, a G1 gate was drawn around the bead singlets on an FSC-H/SSC—H scatter plot. This gated bead population was subsequently analyzed using a histogram plot of FL2-H to evaluate the level of PE staining. Separate markers were drawn around the peaks of the four bead populations (Ml—M4) and the ric mean for FL2 of each bead population was determined. The FL2 geometric mean of each bead was plotted against the lot specific PE/bead values in a log—log plot. Linear regression was performed to obtain a standard curve using the following equation: y = mx + c, with "m" equal to the slope and "0" equal to the y-intercept. ,79, {00265} For PBMC sample analysis, a GI gate was drawn around the positive fluorescent cell population of st on an SSQH/TL-él-ll dot plot. This gated eel! population was subsequently analyzed using a histogram plot of FL2~H to evaluate the level of Pli~laheled antibody staining. The FLQ ric well as mean was determined for each blood ceil sample stained with anti—CDB7-PE or anti—CD20~PE, as unstained control samples. All geometric mean values for 3912 were d t the bead standard curve and values for PE per cell were extrapolated. Since both antibodyl’lfi eonjuga‘tes were at a PEzAb ratio of approximately 1:1, the values for PE per cell correspond to the number of antibodies bound per cell (ABC) value. Experiments were petitioned with duplicate samples for each assay. The mean and standard deviation was determined from several assays for each blood cell population CD37 expression was evaluated in normal blood cells from 4 independent . s were compared to CD20 staining, unstained cells and a non-binding hngG—PE conjugate as controls. example of a typical staining profile of normal B-cells is given in a histograms in Figure l. The average ABC values of 4 different ments for CD37 and CD20 were calculated and listed in Table l.

Table 1: ABC values for CD37 and CD20 expression on human PBMC samples N0 Ab hung-PE CD37 ABC {1320 ABC control control CDl9+Bcells 7744094598 "80 76 CD3+ T cells 2,016 336 74 68 CD56+ NK cells 3,090 264 85 88 CD 1 4+ monocytes 5,244 794 180 215 The highest overall CD37 staining level was found in CDl9+ B—cells at approximately 77,000 ABC. In addition, CD37 staining was seen at low levels in other PBMC populations examined, with CDl4+ monocytes showing CD37 staining at approximately 5,000 ABC, CD56+ NK cells at 3,000 ABC, and CD3+ T cells at 2,000 ABC. Staining with the non—binding hngG-PE control resulted in ABC values of approximately 70 — 90 for B, T and NK cells and approximately 200 for monocytes. In the same 4 donors CD20 expression was evaluated in comparison to CD37. In accordance with published findings, the CD20 staining was restricted mainly to CDl9+ s with an ABC value of approximately 95,000 ABC. The CD20 expression level was just slightly higher than the CD37 expression level. Only minimal CD20 ng was observed in other PBMC populations examined, with CDl4+ monocytes showing CD20 staining at 794 ABC, CD56+ NK cells at 264 ABC and CD3+ T cells at 336 ABC.

This result trates that high CD37 expression is mainly restricted to s in eral blood samples with only minor expression on peripheral T cells, NK cells and monocytes. This is tent with published findings ((Moore et al. 1986, i lmmnnol. l3‘7(9):30l3—8; Sehwartnhlhiez et al, @833, J. hnnmnol., l40l’3)905~9l4). in addition, we found that the CD'S? expression levels on peripheral B-cells is similar to the level of CD20 expression. This expression pattern strongly t that CD37 directed therapies may be a suitable for targeting B—cells in diseases such as B-cell malignancies, autoimmune diseases, inflammatory diseases or other disorders of the immune system analogous to the use of CD20 ed therapies.

Example 2A InvztroBgduleelenonusmreurlfiedl’BMCs {99279} The ability of humanized antibodies to deplete B—eells was measured using in vine assays with human PBMCS according to published studies performed with rlmxiinab {'V'ugmeyster et al.

Cytonietry A. 2003;52{2):ll}l~9 and Vuglneyster et al. int lmmunephannaeol. 2004;4(8):l l l7—i24}.

Alernlnznmab (Campatli) was used as appositive control, since it has been reported to efficiently deplete lymphocytes in viva and in vim: (Hale, Blood. 1933 Oet;62(4)2873—82 and Waldmann, Fllllos Trans R Soc Lond B Biol Sci. 2005 Sep 29;360(lZl-6l):l707-l E).

Fresh buffy coats from healthy donors were obtained from Research Blood Components (Brighton, MA, US) as a source of normal blood cells for all experiments within this study. Buffy coats were prepared by centrifugation of a unit of whole blood and collecting the interface between the plasma and the red blood cells. This unpurified buffy coat contains PBMCs, neutrophils, platelets, red blood cells, and plasma and was used for experiments on the same day it was drawn. Peripheral blood mononuclear cells ) were prepared from buffy coats by standard density gradient centrifugation using Ficoll-Paque as follows. Blood was diluted 1:3 with 1x HBSS containing SmM EDTA and up to mL were added to a 50 mL conical tube. Ten mL of —Paque (GE care) were slowly added to the bottom of each tube. Samples were centrifiaged at 500 x g with no brake at RT for 30 minutes to obtain a layer of PBMCs below the plasma and to remove red blood cells and most granulocytes. The PBMCs were erred to new tubes and washed twice with 1x HBSS containing SmM EDTA by centrifugation at 400 x g for 10 minutes at RT. Staining buffer (1x HBSS, 1% BSA, 0.1% sodium azide) was then used to end the PBMC pellets in the initial blood volume to achieve the original cell density.

Z To assess: the effect ofhuCD37-3. linCDB'?~3~SMCC£§M l huCD37—50~Si\v’lCC— , lnlCD37~50, DMl, ritnximab, alemtnznrna‘n (Carnpath), and "l‘RtHllé on PBMC depletion. 90 at of purified cells. were added to l2 x 75 min polystyrene tubes and incubated with 10 tlL ofa 100 rig/mi, solution of each sample or a ltnlgG lsotyne control antibody for l ltr at 37°C in a humidified 5% (302 incubator. The final antibody (Ab) concentration was ill , in a final volume of ltll} llL in staining butler. Three independent samples were prepared for each treatment. {9&273} To identify populations of PBMCS, all samples were co~stalned immediately alter Ab tion with l0~20 llL of scently labeled Abs ed from, for example. Bl) Biosciences or yi. Anti-CD3-PerCP—Cy5.5 was used to identify T cells, anti-CDl9—APC for B—cells, and anti- CDl4—FITC for monocytes. Staining was d out in a total of 150 uL for 30 min in the dark at RT.

CountBright Absolute Counting Beads (Invitrogen) were vortexed and added to each sample at 50 uL per tube. For PBMC prep samples, cells were washed once with 1 mL staining buffer and centrifuged at 400 x g for 3-5 min. Supernatant was removed with a 1 mL e and cells were resuspended in 500 [AL of 1% formaldehyde in 1x PBS. Samples were stored at 4°C in the dark until acquisition, which was performed within 4 days of sample preparation.

] TreeStar FlowJo software (version PC 7.5) was used for data is. A gate was drawn around the CountBright bead population on an FSC-H vs SSC—H dot plot to determine a total bead count for the sample. To determine the total count for each PBMC population of interest, a separate gate was drawn around the positive fluorescent population on an SSC-H vs FL(x)—H dot plot, where x is the channel of interest. Specifically, a total count for T cells in a sample was found by gating the positive population on an SSC—H vs FL3-H dot plot; for B-cells, the positive population was found on an SSC-H vs FL4-H dot plot; for NK cells, an SSC-H vs FL2-H dot plot was used; for monocytes, an SSC-H vs FLl-H dot plot was used. The ratio of CD19+ cells for B—cells (CD3+ cells for T cells, CD56+ cells for NK cells, or CDl4+ cells for monocytes) relative to beads was determined and multiplied by 100. t depletion was then ated by taking the ratio of the cell to bead ratio in treated samples relative to the cell to bead ratio in isotype control treated s, subtracting this from 1 and multiplying by 100. This corresponds to the ing formula: Percent Depletion = 100 x (1 -cell to bead ratio of treated sample/ cell to bead ratio of control sample). Data for all cell types was analyzed in the same manner.

For two donors tested, treatment of purified PBMC samples with huCD37-3, huCD37-3— SMCC—DMl, huCD37-50 or -50—SMCC-DM1 resulted in approximately 55-70% depletion of B— cells (see Figure 2). There was less than 10% depletion of T cells or monocytes. The B—cell restricted depletion effect tes that this activity is linked to the high CD37 expression on B-cells. In comparison, treatment with the anti-CD20 antibody rituximab resulted in approximately 30—40% depletion of B-cells. Treatment with the anti-CD37 SMIPTM TRU-016 resulted in only 20—30% depletion of B—cells. zumab treatment resulted in depletion of 60—70% of B—cells, 55-65% of T cells and 40-65% of monocytes.

Example 2B Dpseressonseformvztro Brcell d6551€41.91!.HSiIlE.§Z§11£if1§§iRBMQ§ To evaluate the dose-response of the antibodies and conjugates, purified PBMCs from 2 donors were incubated with a 5—fold sample dilution series. Each sample dilution was added at 10 uL per tube to 90 uL of purified cells in triplicate and ted for 1 hour at 37°C in a humidified 5% C02 incubator. The final concentration ranged from 10 ug/mL to 0.13 ng/mL. The same amount of a non— g hngG Ab was used as an isotype control.

For two donors tested, ent of purified PBMC samples with huCD37SMCC-DM1 resulted in a clear dose-response for the B—cell depletion activity (see Figure 3A and B). Incubation with huCD37-3—SMCC-DM1 caused in Vitro depletion of imately 60% of B—cells with an ECSO of 40-75 ng/mL. For an onal donor , treatment of purified PBMC samples with huCDE7-3, huCD37-38, huCD37-50, and huCD37-56 antibodies also resulted in a clear dose—response for the B-cell depletion activity (see Figure 3C). Incubation with these antibodies caused in Vitro depletion of approximately 60- 70% of s with an EC50 of 20-30 ng/mL.

Example 2C InvztroBcelld‘afletlonusmxwmlgblggd The ability of humanized antibodies to deplete B—cells was measured using in Vitro assays with whole blood according to published studies performed with rituximab (Vugmeyster et al. Cytometry A. 2003;52(2)2101-9 and Vugmeyster et al. Int Immunopharmacol. 2004;4(8):l 1 .

Fresh buffs-'7 coats from healthy donors were obtained from Research Blood Components (Brighton, MA, US) as a source of normal blood cells for all experiments within this study. To assess the effect of huCD37-3, huCD37SMCC—DM1, rituximab, alemtuzumab (Campath), and TRU-016 on eral blood cells (PBCs) in a whole blood matrix, 90 pL of whole blood from a buffy coat were incubated with Abs or isotype control as detailed above in a total volume of 100 pL. Three independent s were prepared for each Ab treatment.

To identify populations of blood cells, all samples were co-stained immediately after Ab incubation with 10 - 20 pL of fluorescently d Abs obtained from, for example, BD Biosciences or Miltenyi. Anti-CD3-PerCP-Cy5.5 was used to identify T cells, anti-CDl9-APC for B-cells, anti-CD56— PE for NK cells, and anti-CDl4-FITC for monocytes. Staining was carried out in a total of 150 [1L for 30 min in the dark at RT. CountBright te Counting Beads (Invitrogen #C36950) werepvortexed and added to each sample at 50 pL per tube to allow standardization of cell counts.

] Following cell staining, 2 mL of BD FACS Lysing Solution (BD ences, diluted 1:10 in dH20 according to the manufacturer’s instructions) were added to each sample in order to lyse the RBCs present. Samples were incubated at RT for 15—20 min in the dark, centrifuged at 400 X g for 3—5 min, and resuspended in 500 [1L of 1% formaldehyde in lx PBS. Samples were stored at 4°C in the dark until acquisition, which was performed within 4 days of sample preparation. Samples were acquired on a BD FACSCalibur. Compensation controls were run with each assay to confirm instrument settings. A total of 0 ungated events were acquired for each sample using BD CellQuest re (version 5.2).

TreeStar FlowJo software (version PC 7.5) was used for data analysis as described above.

For one donors tested, treatment of purified PBMC samples with huCD37-3, huCD37 SMCC—DMl, huCD37-50 or —50—SMCC-DM1 ed in approximately 40% depletion of B— cells (see Figure 4). There was less than 10% depletion of T cells, 'NK cells or monocytes. As seen for puréfied PBMCs, the in vitro depletion is restricted to s indicating that the activity is linked to the high CD37 expression on B-cells. In comparison, treatment with the anti—CD20 antibody rituximab or the anti-CD37 SMIPTM TRU—016 resulted in a less than 10% depletion of B-cells. Alemtuzumab treatment resulted in depletion of 40% of B-cells, 80% of T cells, 15% ofNK cells and 20% of monocytes.

Example 2D DovLQl‘OUNOfQEHVfflBLfilliltfiWflé—Lhfillfilfi..lllgfilél 3002283} To evaluate the dose—respense of the antibodies and conjugates? whole bleed from 2 dencrs was incubated with a lG—t‘eld sample on . Each sample dilution was added at it? at. per tube to 90 til, 0f purified cells in triplicate and incubated for '1 hr at 37°C in a humidified 5% C703 incubator. The final concentration ranged from 10 ngij t0 ill L The same amount cf 3 nen~hinding hulgG Ab was used as an isotype central. 300284} Fer two sinners tested, treatment of whole bleed samples with huCD37-3 or huCl'RS7—3— SMCGDMI resulted in a clear dese response for the B~cell ion activity {see Figure 5A and B). ln additlen, huCDBZLSlB was tested fer one (inner and also showed a similar dose se fer the B—cell depletion ty (see Figure SB}. lncnbatioh with huCl‘Bil—E, buCD373~SMCC~DMl er -v50 caused a maximum response at in vitro depletion 0f appreximately 304595 of B~cells with an ECSG of 40420 ng/mL.

In addition to the in vitro experiment described above, the capacity of CD37 antibodies to deplete B cells in vivo can be tested in huCD37 expressing mice (described in Example 3) and, for antibodies that crossreact with macaque CD37, in monkey. e 2E .m,Mm93;itchiinstsl..e«ase studlesusmwhumanPBMCs In vitro cytokine release was measured by ELISpot for IFN-y (Interferon), TNF—a (Tumor Necrosis Factor) and lL-6 leukin-6) using peripheral blood mononuclear cells (PBMCs) from healthy human donors incubated for 18-20 hours with compounds at a concentration of 2.5 ng/mL to 250 pg/mL. The ELISpot method is designed to measure the number of cells ing cytokine by capturing the cytokine onto the assay plate during the entire length of the incubation. In all assays the positive control anti—CD3 dy CD3-2, as well as a negative non—binding isotype hngG control antibody was included. Alemtuzumab (Campath®) and rituximab an®) were used in comparison, since both have been ed to induce cytokine release in patients (Wing. J Clin Invest. 98:2819 (1996) and Winkler, Blood 94:2217—2224 (1999)). The assay conditions were chosen to reflect conditions that are relevant for antibody therapeutics. The highest concentration of 250 pg/mL tested corresponds to the maximum —84- serum tration of an antibody, such as for example the CDZO-directed rituximab, in patient plasma after an on of 10 mg/kg of antibody.

As can be seen in Figures 6 and 7, the positive l anti—CD3 antibody induced release of very high levels of IFN—y, TNF-a and IL-6 with PBMCs from two different . In the same assays, alemtuzumab caused intermediate cytokine release, while rituximab caused moderate cytokine release with PBMCs from two ent donors. In contrast, huCD3 7-3, huCD37—50, huCD37SMCC-DM1 or huCD37SMCC—DM1 did not cause significant cytokine release in our assays.

This underscores the utility of the described CD37-targeting antibodies or conjugates as therapeutics as they combine potent activity, such as B-cell depletion, with a favorable safety profile with respect to cytokine release.

Example 3 1.11_yi129._m9d9.1§.19.§xa1natit11§agixi§:.913.9123]..di:§§t§d_antihgdi§§_9r.,aniuaates B—cell depletion is known to ameliorate autoimmune diseases. In fact, rituximab has been approved for rheumatoid arthritis treatment (Edwards JC et al. Nat Rev Immunol. 6: 119 (2006)). In animal models, B—cell depletion using antibodies against B—cell antigens such as CD20, CD19 and CD79 has been shown to inhibit or rate several autoimmune diseases including systemic lupus erythematosus (SLE), experimental autoimmune encephalomyelitis (EAE; mouse model of multiple sclerosis), type-l diabetes (TlD) and rheumatoid arthritis (RA). The CD37 antigen is expressed at high levels in human B-cells. Therefore, dies or immunoconjugates directed against the CD37 antigen could potentially deplete B-cells and be therefore useful to treat multiple autoimmune diseases.

] To test the utility of CD37 ing antibodies and immunoconjugates to treat human autoimmune es, the activity of such CD37 targeting antibodies and immunoconjugates can be studied in mice using several murine autoimmune disease models.

For example, anti-murine CD37 antibodies can be generated using CD37-knock-out mice or other species such as rat and hamster, and antibodies that deplete B-cell in viva effectively can be selected.

The therapeutic potential of anti-CD37 antibodies can be tested in mouse models representing human autoimmune es, for e, a spontaneous TlD model in NOD mice, a myelin oligodendrocyte glycoprotein (MOG) peptide induced EAE model in wild type C57/Bl6 mice, a en induced rheumatoid tis model in DBA/l mice or a spontaneous systemic lupus erythematosus (SLE) model in MRL/lpr mice. Examples of murine CD37 antibodies and their therapeutic efficacy in various animal models of autoimmune e are ed below. atively, the therapeutic potential of anti-human CD37 antibodies and immunoconjugates can also be tested in murine autoimmune disease models that have been engineered to express the human CD37 antigen. Such human CD37 (huCD37) expressing mice can be generated using standard knock in (Kl) or transgenic (Tg) approaches. For example, to generate huCD37 KI mice, human CD37 cDNA can be inserted into the murine CD37 locus in the C57/Bl6 embryonic stem (ES) cells. The homozygous huCD37 KI mice will express human CD37 cDNA under the regulation of the endogenous murine CD37 promoter, thus the expression pattern of the huCD37 would mimic that of the endogenous muCD37. The different approach es bacterial artificial chromosome (BAC) containing the human CD37 gene that can be randomly inserted into the mouse . This transgenic ch has been used successfully to te huCDZO Tg mice resulting in B-cell c high level expression of the antigen. {(302193} The resulting huCD37 expressing mice based on the C57/Bl6 background can be used to further develop several autoimmune disease model. For examples, MOG peptide immunization in the C57/Bl6 strain background can induces severe EAE in two weeks. In addition, introducing a FcyRIIB knock out phenotype by breeding huCD37 expressing mice with C57/Bl6 FcyRIIB knock out mice should yield a mouse model that spontaneously develop SLE and develop RA upon immunization with en II antigen. Alternatively, backcrossing of the huCD37 expressing C57/Bl6 mice into the NOE?) or MRL/lpr background for 10 generations can provide spontaneous TlD and SLE models, respectively.

Example 4A generatigugfamtianggugllflmgnmlgnal.an.tih9si§:;c_k>ne_2_§_2_:3_ To p proof of concept that CD37 ing dy and immunoconjugate can inhibit autoimmune disease, anti—murine CD37 (muCD37) monoclonal antibodies were generated by immunizing CD37-knock-out C57Bl/6 mice with 300—19, a murine pre-B cell line that endogenously expresses the muCD37 antigen. The immunogen was injected subcutaneously at the dose of 5x106 cells per mouse every 2 weeks for 5 times. Three days before being sacrificed for oma generation, the zed mice received intraperitoneal injection of another dose of antigen. The spleen cells were fused with murine myeloma P3X63Ag8.653 cells (P3 cells) (J. F. Kearney et al. 1979, JImmunol, 123: 550) at ratio of 1 P3 cells: 3 spleen cells according to standard procedure. The fused cells were ed in RPMI-1640 ion medium containing hypoxanthine-aminopterin—thymidine (HAT) (Sigma Aldrich) in % C02 incubator at 37°C until hybridoma clones were ready for antibody screening.

Screening was done using flow cytometric binding assay with spleen cells from wild type mice and CD37-knockout mice. Tie spleen cells were counterstained with anti—CD45R (8220) antibody to identify B cells that constitutively express CD37 antigen. The hybridomas producing antibody that bound the wild type, but not nock-out, B cells were subcloned by limiting dilution. One stable subclone (clone 252-3) was obtained. The 252—3 hybridoma was expanded in low IgG serum containing media and the antibody was purified using standard methods with protein A/G chromatography. -86— Example 4B The purified 252-3 monoclonal antibody was identified as a mouse IgG2a with lsoStrip mouse monoclonal antibody isotypirig kit (Roche Diagnostics Corporation, Indianapolis, IN). To determine the binding affinity to the muCD37 n, various trations of 252-3 antibody were incubated with 300-19 cells, a murine pre-B cell line that expresses the muCD37 n, for 30 minutes at 4°C. Cells were then washed and counterstained with anti mulgG—PE conjugate (Jackson research, West Grove, PA) for 30 minutes at 4°C. The cells were finally , fixed in formalin and analyzed by flow cytometry using a FACSarray (BD Bioscience, San Jose, CA). The flow cytometry data were analyzed using FlowJo (Tree Star Inc., Ashland, OR) and the geometric mean fluorescence intensity was d against the dy concentration in a semi-log plot (Figure 8). A dose-response curve was generated by non-linear regression and the ECSO value of the curve, which coréesponds to the apparent dissociation constant (Kd) of the antibody, was calculated using GraphPad Prism (GraphPad Software Inc., La Jolla, CA). It was found that the Kd of the 252—3 antibody was 14 nM.

In contrast, the 252-3 antibody did not bind to human tumor cells expressing the human CD37 n.

The 252-3 antibody was then used as a surrogate antibody in murine autoimmune disease models to demonstrate the therapeutic potential of a CD37—targeting antibody for the ent of autoimmune diseases (Examples 5-7).

Example 5 i ..»..<.‘....._ ] Experémental autoimmune encephalomyelitis (EAE) is an animal model of inflammatory demyelinating disease of the central nervous system (CNS), including le sclerosis in human.

Murine EAE is ly induced by zation of spinal cord homogenates, brain extracts, or CNS protein such as myelin protein or peptide, followed by injection of pertussis toxin to break down the blood-brain barrier and allow immune cells access to the CNS tissue. This immunization leads to multiple small disseminated lesions of demyelination in the brain and spinal cord, causing tail paralysis followed by limb paralysis.

To test the activity of anti-muCD37 antibody in the EAE model, we first studied the capacity of the 252—3 antibody to deplete B cells in viva. C57Bl/6 mice were injected intraperitoneally with 25 mg/kg of 252—3 antibody or onal muréne IgG (Jackson Immunoresearch, West Grove, PA) as a control. Peripheral blood was collected at different time points and analyzed for B and T cell levels by flow cytometry, Allophycocyanin (APC)—conjugated anti—mouse CD45R (B220) antibody (ebioscience, -37..

San Diego, CA) and fluorescein isothiocyanate —conjugated anti CD38 dy (ebioscience, San Diego, CA) were used to stain B and T cell populations, tively. B cell depletion was assessed by calculating the ratio of B to T cells for each sample and the B/T ratio was normalized by setting the average B/T ratio of murine IgG—treated s to 100%. The normalized B/T cell ratio was plotted mngG control mice and 252-3 dy treated mice (Figure 9A). The result show that the B cell level of the mice treated with 252-3 antibody was rapidly reduced within a few hours after the antibody injection.

The B cell depletion reached ~70% at 3h and peaked at day 3 (> 95%). After day 3, the B cell level slowly increased and reached ~60% of the normal level at day 14. This data ts that the 252—3 antibody can rapidly and efficiently deplete peripheral blood B cells, and this effect was sustained for at least 7 days after the antibody injection.

The second study tested the capacity of 252-3 antibody to inhibit EAE. In this study, EAE was induced in C57Bl/6 mice by subcutaneous immunization of MOG35_55 peptide emulsified in complete Freund’s adjuvant (EAE kit from Hooke Laboratories, Lawrence, MA) into the upper and lower back at day 0 and two eritoneal injections of pertussis toxin at 21": and 24h after antigen immunization. Mice were checked for EAE signs daily starting on day 7 after immunization. The disease severity was scored on a scale of 0 to 5 using the following criteria: Cl1nlcal Observations 0 ENO s changes1n motor functions of the mouse in comparison to non--immunized mice When picked up by the tail, the tail has n and1s erect. Hind legs are usually spread apartE Whenthe mouse is walklng, there1s no gait or head t1lt1ng l Limp tail.

When the mouse is picked up by tail, instead of being erect, the whole tail drapes over your 3 finger. 2 Limp tail and weakness of hind legs.

When the mouse is picked up by tail, legs are not spread apart, but held closer together. When the mouse is observed when walking, it has clearly apparent wobbly walk. i E 3 Limp tail and complete paralysis of hind legs (most common) E Limb tail with paralysis of one front and one hind leg.

OR, ALL of: *8 Severe head tilting 8 Walking only along the edges of the cage a Pushing against the cage wall 1* Spinnmg when p1ckedupbythe tail "88- 4 Limp tail, complete hind leg and partial front leg paralysis.

Mouse is minimally moving around the cage but appears alert and feeding. Usually, euthanasia is recommended after the mouse scores level 4 for 2 days. When the mouse is euthanized because of severe paralysis, a score of 5 is entered for that mouse for the rest of the experiment.

Complete hind and front leg paralysis, no movement around the cage. 3 OR, t Mouse is spontaneously rolling in the cage. I OR, : Mouse is found dead due to paralysis. i If mouse is alive, euthanize the mouse immediately if it scores 5. Once mouse scored 5, the same score is entered for all the days for the rest ofthe experiment. =iii‘dsiib‘‘1‘‘'1‘‘‘‘'"''Ali'iii{£5'é"é'i'e'fr'i'eifc'l'"iESEBB?‘v‘‘Ei;£,§ii"§ETf‘iiXiawwbethErT"13313"1‘3?£121};"é‘fiéFEiiiiii;EH"iiiiiiiii‘ri1221i}’cSiIKil the disease onset, mice were randomized and the 252-3 antibody or polyclonal mulgG was ed once intraperitoneally at a 25 mg/kg dose. A total of 10 mice were enrolled for each group. At the end of the study (18 days after the disease onset), the data were synchronized based on the day of disease onset for each mouse. The disease progression plot (Figure 9B) shows that mice from both groups had relapsing- remitting form of EAE. During the first wave of clinical symptoms, the control mice reached the mean of 3 while the mice treated with 252-3 dy had a mean of 2. The difference in disease severity between these two groups was sustained for more than 2 weeks after the disease onset. Taken er, this data suggests that the 252-2 antibody ent rapidly depletes the B cell population and alleviates EAE.

Example 6 Anti-muQQ§.Z_m9n9919nraLanti.b9d§i__irthitzit§Eritreidiabe esmNODmlce Type-l diabetes (TlD) or juvenile es or insulin-dependent diabetes millitus (IDDM) is caused by mmune reaction against n—producing pancreatic beta cells. Destruction of beta cells reduces insulin production and increases glucose level that produces various clinical symptoms. TlD incidence in Northern Europe and the US is between 8 and 17/ 100,000. Insulin supplement is the most common treatment of the disease.

Non-obese diabetic (NOD) mice spontaneously develop TlD and have been widely used to model the human disease. In NOD mice, the e starts with leukocytic infiltration of the pancreatic islets (called tis) as early as 4 weeks of age. The insulitis progresses rapidly, leading to destruction of pancreatic islets and diabetes ng at 12-15 weeks of age. B cell depletion using anti-CD20 antibody in the early stage of insulitis has been reported to delay the disease onset (Hu et al., J Clin lnves. 117, 3857 ), suggesting that B cells play a critical role in the e pathogenesis in NOD mice.

To test the activity of anti-muCD37 antibody, the 252—3 antibody was injected into six female NOD mice intraperitoneally at 25 mg/kg every 10 days for a total of 4 injections starting at 5 weeks of age (n=6). The control mice (n=6) were injected with polyclonal murine IgG (Jackson Immunoresearch, West Grove, PA). Three days after the last ion, the B and T cell levels in eral blood were examined by flow cytometry. Allophycocyanin (APC)—conjugated anti—mouse CD45R (B220) antibody (ebioscience, San Diego, CA) and fluorescein isothiocyanate (FITC)-conjugated anti CD38 antibody (ebioscience, San Diego, CA) were used to stain B and T cell tions, respectively. The B/T cell ratio was normalized to murine IgG control treated samples as described above and the normalized B/T cell ratio was plotted for mngG control mice and 252-3 antibody treated mice (Figure 10A). The results show that the B cell level of the mice treated with 252-3 antibody was significantly reduced as compared to the control mice, suggesting that the 252-3 antibody efficiently depletes peripheral blood B cells in NOD mice. To examine the effect of B cell depletion by anti-muCD37 antibody, blood e level was measured weekly starting at 12 weeks of age. Mice with blood glucose level 2 250 mg/dL in two consecutive weeks are ered diabetic. The data in Figure 10B shows that the control mice started to develop diabetes on week 15 and 83% of the mice had diabetes on week 22. In contrast, the mice treated with 252-3 antibody d to develop diabetes on week 17 and only 50% of the mice were diabetic on week 27. This data shows that treatment of 252—3 antibody efficiently depletes B cells in NOD mice, delays the onset of es and significantly reduces the e incidence.

Example 7 Collagen-induced arthritis (CIA) is an animal model of rheumatoid arthritis (RA) that is widely used to investigate disease enesis and to validate therapeutic targets. Arthritis is normally induced in mice or rats by zation with gous or heterologous type II collagen in adjuvant.

This immunization elicits a robust T- and B- cell response to the antigen leading to proliferative synovitis with infiltration of rphonuclear and mononuclear cells, pannus formation, cartilage ation, bone erosion and fibrosis.

Since different mouse strains have different susceptibility to antibody-mediated B cell depletion (Ahuja et al., J. Immunol, 179: 361 (2007)), to test the activity of anti—muCD37 antibody in CIA model, we first studied the capacity of the 252-3 antibody to deplete B cells in DBA/ 1 mice. Mice were injected intraperitoneally with 25 mg/kg of 252-3 antibody or polyclonal murine IgG (Jackson Immunoresearch, West Grove, PA) as control. Peripheral blood was collected at ent time points and analyzed for B and T cell levels by flow cytometry. Allophycocyanin (APC)—conjugated anti-mouse CD45R (B220) antibody (ebioscience, San Diego, CA) and fluorescein isothiocyanate (FITC)—conjugated anti CD38 antibody (ebioscience, San Diego, CA) were used to stain B and T cell populations, respectively. The normalized B/T cell ratio was was calculated as described above and compared between the mngG control mice and 252—3 antibody treated mice (Figure 11A). The result show that the 252—3 antibody significantly reduced the peripheral blood B cell level to ~20% and ~8% in l and 3 days after the antibody injection, and this low B cell level was ined at 7 days after the antibody injection. This data suggests that the 252-3 antibody can rapidly and efficiently deplete peripheral blood B cells, and this effect was sustained for at least 7 days after the antibody ion.

The second study tests the capacity of 252-3 antibody to inhibit CIA. In this study, CIA was induced in DBA/1 mice by subcutaneous immunization of chicken collagen/CFA ete Freund’s adjuvant) on day 0 and chicken en/IFA (incomplete Freund’s nt) on day 21 (Hooke Laboratories, Lawrence, MA). Mice were checked for CIA signs daily starting on day 21 after immunization. The CIA ty was scored on a scale of 0 to 16 (based on a score of 0 to 4 for each paw) using the following criteria: Clinics", servatlons Normalpaw,"_ fiuOneto__e__ 1nflamed and swollen More than one toe, butnot entirepaw, inflamed and swollen Mild swelhnv repaw\ __________ ntirepaw1nflamedand swollen 4 Very inflamedandswollen paw or ankylosed paw."Ifthepaw 1 isankylosedthemouse grip thewiretopofthepage..." ] At the onset of arthritis symptoms, mice were randomized into two groups and injected with the 252—3 antibody or polyclonal mulgG intraperitoneally at 10 mg/kg dose at three consecutive days. A total of 12 mice were enrolled for each group. At the end of the study (21 days after the disease onset), the data were synchronized based on the day of disease onset for each mouse. The disease progression plot (Figure 11B) shows that the disease severity in control mice sed rapidly from mean score of 2 at day 1 to 9.5 at day 7. In contrast, the disease in mice d with the 252—3 antibody progressed significantly slower with mean score of 4.4 at day 7. Altogether, this data suggests that the 252-2 antibody treatment significantly depletes the B cell population and alleviates CIA.

In conclusion, the above experiments using a surrogate anti—muCD37 antibody provide evidence that a CD37—targeting antibody, or an immunoconjugate that includes a CD37 dy, can t autoimmune diseases in animal models.

**** It is to be appreciated that the Detailed Description section, and not the Abstract section, is intended to be used to interpret the claims. The Abstract Abstract may set forth one or more but not all "9].. exemplary embodiments of the present invention as contemplated by the ors, and thus, is not intended to limit the t invention and the appended claims in any way.

The present ion has been described above with the aid of functional building blocks illustrating the implementation of specified functions and relationships thereof The boundaries of these functional building blocks have been arbitrarily defined herein for the convenience of the description.

Alternate boundaries can be defined so long as the specified functions and relationships f are riately performed.

The foregoing description of the specific embodiments will so fully reveal the l nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for varéous applications such specific embodiments, without undue experimentation, without departing from the l concept of the present invention. Therefore, such tions and modifications are intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented . It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.

The breadth and scope of the present invention should not be limited by any of the above- described exemplary embodiments, but should be defined only in accordance with the ing claims and their equivalents.

All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each independent publication, patent, or patent application was specifically and individually indicated to be orated by reference.

Other embodiments of the invention as described herein are defined in the following paragraphs: 1. A method for depleting a B-cell comprising contacting a population of cells comprising a noncancerous B-cell with an antibody or antigen binding fragment thereof that specifically binds to CD37, n said antibody or fragment thereof is capable of inducing apoptosis in vitro in the absence of a linking agent. 2. A method for treating a patient having an mune or inflammatory disease sing administering to said patient a eutically effective amount of an antibody or antigenbinding fragment thereof that specifically binds to CD37, wherein said antibody or fragment thereof is capable of inducing sis in vitro in the absence of a cross-linking agent. 3. The method of paragraph 1 or 2, wherein said antibody or antigen-binding fragment f is also capable of inducing complement dependent cytotoxicity (CDC). 4. The method of any of aphs 1-3, wherien said dy or antigen-binding fragment thereof is also capable of inducing antibody dependent cell mediated cytotoxicity (ADCC).

. A method for depleting a B-cell comprising contacting a population of cells comprising a noncancerous B-cell with an antibody or antigen g fragment f that specifically binds to the same CD37 epitope as an antibody selected from the group consisting of: (a) an antibody comprising the polypeptide of SEQ ID NO:55 and the polypeptide of SEQ ID NO:72; (b) an antibody comprising the polypeptide of SEQ ID NO:56 and the polypeptide of SEQ ID NO:73; (c) an dy comprising the polypeptide of SEQ ID NO:57 and the polypeptide of SEQ ID NO:74; (d) an antibody comprising the ptide of SEQ ID NO:58 and the polypeptide of SEQ ID NO:74; (e) an antibody comprising the polypeptide of SEQ ID NO:59 and the ptide of SEQ ID NO:75; (f) an antibody comprising the polypeptide of SEQ ID NO:60 and the polypeptide of SEQ ID NO:76; (g) an antibody comprising the polypeptide of SEQ ID NO:61 and the polypeptide of SEQ ID NO:77; (h) an antibody comprising the polypeptide of SEQ ID NO:62 and the polypeptide of SEQ ID NO:78; (i) an antibody comprising the polypeptide of SEQ ID NO:63 and the polypeptide of SEQ ID NO:79; (j) an antibody comprising the polypeptide of SEQ ID NO:64 and the polypeptide of SEQ ID NO:80; (k) an antibody comprising the polypeptide of SEQ ID NO:65 and the polypeptide of SEQ ID NO:81; (l) an antibody sing the polypeptide of SEQ ID NO:66 and the polypeptide of SEQ ID NO:82; (m) an antibody comprising the polypeptide of SEQ ID NO:67 and the polypeptide of SEQ ID NO:83; (n) an antibody comprising the polypeptide of SEQ ID NO:68 and the polypeptide of SEQ ID NO:84; (o) an antibody comprising the polypeptide of SEQ ID NO:69 and the polypeptide of SEQ ID NO:85; (p) an antibody comprising the polypeptide of SEQ ID NO:70 and the polypeptide of SEQ ID NO:86; (q) an antibody comprising the polypeptide of SEQ ID NO:71 and the polypeptide of SEQ ID NO:87; (r) an antibody comprising the polypeptide of SEQ ID NO:177 and the polypeptide of SEQ ID NO:178. 6. A method for treating a patient having an autoimmune or inflammatory disease comprising administering to said t a therapeutically effective amount of an antibody or antigen- binding fragment thereof that specifically binds to the same CD37 e as an dy selected from the group consisting of: (a) an antibody comprising the polypeptide of SEQ ID NO:55 and the polypeptide of SEQ ID NO:72; (b) an antibody comprising the polypeptide of SEQ ID NO:56 and the polypeptide of SEQ ID NO:73; (c) an antibody comprising the polypeptide of SEQ ID NO:57 and the polypeptide of SEQ ID NO:74; (d) an dy comprising the polypeptide of SEQ ID NO:58 and the polypeptide of SEQ ID NO:74; (e) an antibody comprising the polypeptide of SEQ ID NO:59 and the polypeptide of SEQ ID NO:75; (f) an antibody comprising the ptide of SEQ ID NO:60 and the polypeptide of SEQ ID NO:76; (g) an antibody comprising the polypeptide of SEQ ID NO:61 and the polypeptide of SEQ ID NO:77; (h) an antibody comprising the polypeptide of SEQ ID NO:62 and the polypeptide of SEQ ID NO:78; (i) an antibody comprising the polypeptide of SEQ ID NO:63 and the polypeptide of SEQ ID NO:79; (j) an antibody comprising the polypeptide of SEQ ID NO:64 and the polypeptide of SEQ ID NO:80; (k) an antibody comprising the polypeptide of SEQ ID NO:65 and the polypeptide of SEQ ID NO:81; (l) an antibody comprising the polypeptide of SEQ ID NO:66 and the polypeptide of SEQ ID NO:82; (m) an antibody sing the polypeptide of SEQ ID NO:67 and the polypeptide of SEQ ID NO:83; (n) an antibody comprising the polypeptide of SEQ ID NO:68 and the polypeptide of SEQ ID NO:84; (o) an dy comprising the polypeptide of SEQ ID NO:69 and the polypeptide of SEQ ID NO:85; (p) an antibody comprising the polypeptide of SEQ ID NO:70 and the polypeptide of SEQ ID NO:86; (q) an dy comprising the polypeptide of SEQ ID NO:71 and the polypeptide of SEQ ID NO:87; and (r) an antibody comprising the polypeptide of SEQ ID NO:177 and the polypeptide of SEQ ID NO:178. 7. The method of paragraph 5 or 6, wherein the antibody or antigen-binding fragment thereof itively inhibits an antibody selected from the group consisting of: (a) an antibody comprising the polypeptide of SEQ ID NO:55 and the polypeptide of SEQ ID NO:72; (b) an antibody comprising the ptide of SEQ ID NO:56 and the polypeptide of SEQ ID NO:73; (c) an antibody comprising the polypeptide of SEQ ID NO:57 and the polypeptide of SEQ ID NO:74; (d) an antibody comprising the polypeptide of SEQ ID NO:58 and the polypeptide of SEQ ID NO:74; (e) an antibody sing the polypeptide of SEQ ID NO:59 and the polypeptide of SEQ ID NO:75; (f) an antibody comprising the polypeptide of SEQ ID NO:60 and the polypeptide of SEQ ID NO:76; (g) an antibody comprising the polypeptide of SEQ ID NO:61 and the polypeptide of SEQ ID NO:77; (h) an antibody comprising the polypeptide of SEQ ID NO:62 and the polypeptide of SEQ ID NO:78; (i) an antibody comprising the polypeptide of SEQ ID NO:63 and the polypeptide of SEQ ID NO:79; (j) an antibody comprising the polypeptide of SEQ ID NO:64 and the ptide of SEQ ID NO:80; (k) an dy comprising the polypeptide of SEQ ID NO:65 and the polypeptide of SEQ ID NO:81; (l) an antibody comprising the ptide of SEQ ID NO:66 and the polypeptide of SEQ ID NO:82; (m) an antibody comprising the polypeptide of SEQ ID NO:67 and the polypeptide of SEQ ID NO:83; (n) an dy comprising the polypeptide of SEQ ID NO:68 and the polypeptide of SEQ ID NO:84; (o) an antibody comprising the polypeptide of SEQ ID NO:69 and the polypeptide of SEQ ID NO:85; (p) an antibody comprising the polypeptide of SEQ ID NO:70 and the polypeptide of SEQ ID NO:86; (q) an antibody comprising the polypeptide of SEQ ID NO:71 and the polypeptide of SEQ ID NO:87; (r) an antibody comprising the polypeptide of SEQ ID NO:177 and the polypeptide of SEQ ID NO:178. 8. A method for depleting a B-cell sing contacting a population of cells comprising noncancerous B-cell with an antibody or antigen-binding fragment thereof ed by hybridoma selected from the group ting of ATCC t Designation PTA-10664, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation 665, ted with the ATCC on February 18, 2010, ATCC Deposit Deisgnation PTA-10666, ted with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10667 deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10668, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10669, deposited with the ATCC on February 18, 2010, and ATCC Deposit Designation PTA-10670, deposited with the ATCC on February 18, 2010. 9. A method for treating a patient having an autoimmune or inflammatory disease comprising administering to said patient a therapeutically effective amount of an dy or antigenbinding fragment thereof ed by hybridoma selected from the group consisting of ATCC Deposit Designation PTA-10664, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10665, deposited with the ATCC on February 18, 2010, ATCC Deposit Deisgnation PTA-10666, deposited with the ATCC on February 18, 2010, ATCC Deposit ation PTA-10667 deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10668, deposited with the ATCC on February 18, 2010, ATCC Deposit Designation PTA-10669, deposited with the ATCC on February 18, 2010, and ATCC Deposit Designation PTA-10670, deposited with the ATCC on February 18, 2010.

. A method for depleting a B-cell comprising contacting a population of cells sing a noncancerous B-cell with an antibody or antigen-binding nt thereof that specifically binds to CD37, wherein said antibody ses polypeptide sequences selected from the group consisting of: (a) SEQ ID NOs: 4, 5, and 6 and SEQ ID NOs: 28, 29, and 30; (b) SEQ ID NOs: 7, 8, and 9 and SEQ ID NOs: 31, 32, and 33; (c) SEQ ID NOs: 10, 11, and 12 and SEQ ID NOs: 34, 35, and 36; (d) SEQ ID NOs: 13, 14, and 15 and SEQ ID NOs: 37, 38, and 39; (e) SEQ ID NOs: 13, 14, and 15 and SEQ ID NOs: 37, 40, and 39; (f) SEQ ID NOs: 16, 17, and 18 and SEQ ID NOs: 41, 42, and 43; (g) SEQ ID NOs: 19, 20, and 21 and SEQ ID NOs: 44, 45, and 46; (h) SEQ ID NOs: 19, 20, and 21 and SEQ ID NOs: 44, 47, and 46; (i) SEQ ID NOs: 22, 23, and 24 and SEQ ID NOs: 48, 49, and 50; (j) SEQ ID NOs: 22, 23, and 24 and SEQ ID NOs: 48, 51, and 50; (k) SEQ ID NOs: 25, 26, and 27 and SEQ ID NOs: 52, 53, and 54; (l) SEQ ID NOs: 171, 172, and 173 and SEQ ID NOs: 174, 175, and 176; (m) SEQ ID NOs: 171, 181, and 173 and SEQ ID NOs: 174, 175, and 176; and (n) variants of (a) to (m) comprising 1, 2, 3, or 4 conservative amino acid substitutions. 11. A method for treating a patient having an autoimmune or inflammatory disease comprising administering to said t a therapeutically effective amount of an antibody or antigenbinding fragment thereof with an antibody or antigen-binding fragment thereof that specifically binds to CD37, wherein said antibody comprises polypeptide ces selected from the group consisting of: (a) SEQ ID NOs: 4, 5, and 6 and SEQ ID NOs: 28, 29, and 30; (b) SEQ ID NOs: 7, 8, and 9 and SEQ ID NOs: 31, 32, and 33; (c) SEQ ID NOs: 10, 11, and 12 and SEQ ID NOs: 34, 35, and 36; (d) SEQ ID NOs: 13, 14, and 15 and SEQ ID NOs: 37, 38, and 39; (e) SEQ ID NOs: 13, 14, and 15 and SEQ ID NOs: 37, 40, and 39; (f) SEQ ID NOs: 16, 17, and 18 and SEQ ID NOs: 41, 42, and 43; (g) SEQ ID NOs: 19, 20, and 21 and SEQ ID NOs: 44, 45, and 46; (h) SEQ ID NOs: 19, 20, and 21 and SEQ ID NOs: 44, 47, and 46; (i) SEQ ID NOs: 22, 23, and 24 and SEQ ID NOs: 48, 49, and 50; (j) SEQ ID NOs: 22, 23, and 24 and SEQ ID NOs: 48, 51, and 50; (k) SEQ ID NOs: 25, 26, and 27 and SEQ ID NOs: 52, 53, and 54; and (l) variants of (a) to (k) comprising 1, 2, 3, or 4 vative amino acid substitutions. 12. The method of paragraph 10 or 11, n the antibody or antigen-binding fragment thereof comprises polypeptide sequences that are at least 90% identical to polypeptide sequences selected from the group consisting of: (a) SEQ ID NO:55 and SEQ ID NO:72; (b) SEQ ID NO:56 and SEQ ID NO:73; (c) SEQ ID NO:57 and SEQ ID NO:74; (d) SEQ ID NO:58 and SEQ ID NO:74; (e) SEQ ID NO:59 and SEQ ID NO:75; (f) SEQ ID NO:60 and SEQ ID NO:76; (g) SEQ ID NO:61 and SEQ ID NO:77; (h) SEQ ID NO:62 and SEQ ID NO:78; (i) SEQ ID NO:63 and SEQ ID NO:79; (j) SEQ ID NO:64 and SEQ ID NO:80; (k) SEQ ID NO:65 and SEQ ID NO:81; (l) SEQ ID NO:66 and SEQ ID NO:82; (m) SEQ ID NO:67 and SEQ ID NO:83; (n) SEQ ID NO:68 and SEQ ID NO:84; (o) SEQ ID NO:69 and SEQ ID NO:85; (p) SEQ ID NO:70 and SEQ ID NO:86; (q) SEQ ID NO:71 and SEQ ID NO:87; and (r) SEQ ID NO:177 and SEQ ID NO:178. 13. The method of paragraph 12, wherein the polypeptide sequences are at least 95% identical to polypeptide sequences selected from the group consisting of: (a) SEQ ID NO:55 and SEQ ID NO:72; (b) SEQ ID NO:56 and SEQ ID NO:73; (c) SEQ ID NO:57 and SEQ ID NO:74; (d) SEQ ID NO:58 and SEQ ID NO:74; (e) SEQ ID NO:59 and SEQ ID NO:75; (f) SEQ ID NO:60 and SEQ ID NO:76; (g) SEQ ID NO:61 and SEQ ID NO:77; (h) SEQ ID NO:62 and SEQ ID NO:78; (i) SEQ ID NO:63 and SEQ ID NO:79; (j) SEQ ID NO:64 and SEQ ID NO:80; (k) SEQ ID NO:65 and SEQ ID NO:81; (l) SEQ ID NO:66 and SEQ ID NO:82; (m) SEQ ID NO:67 and SEQ ID NO:83; (n) SEQ ID NO:68 and SEQ ID NO:84; (o) SEQ ID NO:69 and SEQ ID NO:85; p) SEQ ID NO:70 and SEQ ID NO:86; (q) SEQ ID NO:71 and SEQ ID NO:87; and (r) SEQ ID NO:177 and SEQ ID NO:178. 14. The method of paragraph 13, wherein the polypeptide sequences are at least 99% identical to ptide sequences selected from the group consisting of: (a) SEQ ID NO:55 and SEQ ID NO:72; (b) SEQ ID NO:56 and SEQ ID NO:73; (c) SEQ ID NO:57 and SEQ ID NO:74; (d) SEQ ID NO:58 and SEQ ID NO:74; (e) SEQ ID NO:59 and SEQ ID NO:75; (f) SEQ ID NO:60 and SEQ ID NO:76; (g) SEQ ID NO:61 and SEQ ID NO:77; (h) SEQ ID NO:62 and SEQ ID NO:78; (i) SEQ ID NO:63 and SEQ ID NO:79; (j) SEQ ID NO:64 and SEQ ID NO:80; (k) SEQ ID NO:65 and SEQ ID NO:81; (l) SEQ ID NO:66 and SEQ ID NO:82; (m) SEQ ID NO:67 and SEQ ID NO:83; (n) SEQ ID NO:68 and SEQ ID NO:84; (o) SEQ ID NO:69 and SEQ ID NO:85; p) SEQ ID NO:70 and SEQ ID NO:86; (q) SEQ ID NO:71 and SEQ ID NO:87; and (r) SEQ ID NO:177 and SEQ ID NO:178.

. The method of any one of paragraphs 1-14, wherein said antibody or antigen binding fragment thereof is murine, man, humanized, ic, resurfaced, or human. 16. The method of any one of paragraphs 5-15, wherein said antibody or antibody fragment is capable of inducing apoptosis of a cell sing CD37 in vitro in the absence of cross-linking agents. 17. The method of any one of paragraphs 5-15, wherein said antibody or antigen g fragment is capable of inducing complement dependent cytotoxicity (CDC). 18. The method of any one of paragraphs 5-15, wherein said antibody is capable of inducing antibody dependent cell mediated cytotoxicity (ADCC). 19. A method for depleting a B-cell comprising ting a population of cells comprising a noncancerous B-cell with a human or humanized antibody or antigen binding fragment thereof that specifically binds to CD37, wherein said antibody or fragment thereof is capable of inducing ment dependent cytotoxicity (CDC).

. A method for ng a t having an autoimmune or inflammatory disease comprising administering to said patient a therapeutically effective amount of a human or humanized antibody or antigen binding fragment f that specifically binds to CD37, wherein said antibody or fragment thereof is capable of inducing complement dependent cytotoxicity (CDC). 21. The method of paragraph 19 or 20, wherein said human or humanized dy or antigen binding fragment thereof is also e of inducing inducing apoptosis in vitro in the absence of a cross-linking agent. 22. The method of any one of paragraphs 20-21, wherein said human or humanized antibody or antigen binding fragment thereof is also capable of inducing antibody dependent cell ed cytotoxicity (ADCC). 23. The method of any one of paragraphs 1-22 wherein said antibody binds to human CD37 and macaque CD37. 24. The method of any one of paragraphs 1-22, which is a full length antibody.

. The method of any one of paragraphs 1-22, which is an antigen-binding fragment. 26. The method of any one of paragraphs 1-22, wherien said antibody or antigen-binding fragment thereof comprises a Fab, Fab', F(ab')2, Fd, single chain Fv or scFv, disulfide linked Fv, V-NAR domain, IgNar, intrabody, IgGΔCH2, minibody, F(ab')3,tetrabody, triabody, diabody, singledomain antibody, DVD-Ig, Fcab, mAb2, (scFv)2, or scFv-Fc. 27. The method of any one of paragraphs 1-26, wherein the antibody or antigen-binding fragment thereof is linked via a linker (L) to a cytotoxic agent (C) to form an conjugate. 28. A method for depleting a B-cell comprising ting a population of cells comprising a noncancerous B-cell with a composition comprising an immunoconjugate having the formula (A) - (L) - (C), n: (A) is an antibody or antigen binding fragment that specifically binds to CD37; (L) is a non-cleavable linker; and (C) is a cytotoxic agent; and wherein said linker (L) links (A) to (C). 29. A method for treating a patient having an autoimmune or inflammatory disease comprising stering to said patient a therapeutically effective amount of a composition comprising an immunoconjugate having the formula (A) - (L) - (C), wherein: (A) is an antibody or antigen binding nt that specifically binds to CD37; (L) is a non-cleavable linker; and (C) is a xic agent; and wherein said linker (L) links (A) to (C).

. A method for depleting a B-cell comprising contacting a population of cells comprising a noncancerous B-cell with a composition comprising an immunoconjugate having the formula (A) - (L) - (C), wherein: (A) is an antibody or antigen binding fragment that specifically binds to CD37; (L) is a linker; and (C) is a maytansinoid; and wherein said linker (L) links (A) to (C). 31. A method for treating a patient having an autoimmune or matory disease comprising administering to said t a eutically effective amount of a composition comprising an immunoconjugate having the formula (A) - (L) - (C), wherein: (A) is an antibody or antigen binding fragment that specifically binds to CD37; (L) is a linker; and (C) is a maytansinoid; and wherein said linker (L) links (A) to (C). 32. The method of paragraph 30 or 31, n the linker is a non-cleavable linker. 33. The method of any one of aphs 27-32, wherein the immunoconjugate further comprises a second (C). 34. The method of paragraph 33, n the immunoconjugate further comprises a third (C).

. The method of paragraph 34, wherein the immunoconjugate further comprises a fourth (C). 36. The method of one of paragraphs 27-32, wherein the immunoconjugate comprises 2-6 (C). 37. The method of paragraph 36, n the immunoconjugate comprises 3-4 (C). 38. The method of anyone of paragraphs 27-37, wherein said linker is selected from the group consisting of a cleavable linker, a non-cleavable linker, a hydrophilic linker, and a dicarboxylic acid based linker. 39. The method of any one of paragraphs 27-38, wherein said linker is selected from the group ting of: N-succinimidyl 4-(2-pyridyldithio)pentanoate (SPP); N-succinimidyl 4-(2- pyridyldithio)butanoate (SPDB) or N-succinimidyl 4-(2-pyridyldithio)sulfobutanoate (sulfo- SPDB); N-succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC); N- sulfosuccinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (sulfoSMCC); N-succinimidyl- 4-(iodoacetyl)-aminobenzoate (SIAB); and N-succinimidyl-[(N-maleimidopropionamido)- tetraethyleneglycol] ester (NHS-PEG4-maleimide). 40. The method of paragraph 39, wherein said linker is N-succinimidyl-[(N- maleimidopropionamido)-tetraethyleneglycol] ester (NHS-PEG4-maleimide). 41. The method of any one of aphs 27-29 and 33-40, wherein said cytotoxic agent is selected from the group consisting of a maytansinoid, maytansinoid analog, doxorubicin, a ed doxorubicin, benzodiazepine, taxoid, CC-1065, CC-1065 , duocarmycin, duocarmycin analog, calicheamicin, dolastatin, dolastatin analog, aristatin, tomaymycin tive, and ycin derivative or a prodrug of the agent. 42. The method of paragraph 41, wherein said cytotoxic agent is a maytansinoid. 43. The method of any one of paragraphs 30-32 or 42, wherein said cytotoxic agent is N(2')- deacetyl-N(2')-(3-mercaptooxopropyl)-maytansine (DM1) or N(2')-deacetyl-N2-(4-mercapto- 4-methyloxopentyl)-maytansine (DM4). 44. The method of any one of paragraphs 27-43, wherein the composition comprising an immunoconjugate comprises multiple cytotoxic agents (C) with an e of about 3 to about 4 (C) per (A). 45. The method of paragraph 44, n the immunoconjugates have an average of about 3.5 ± 0.5 (C) per (A). 46. The method of paragraph 45, wherein the immunoconjugates have an average of about 3.5 (C) per (A). 47. The method of any one of 1-46, wherein said antibody or antigen-binding fragment is capable of depleting B-cells. 48. The method of any one of aphs 1-47, wherein said dy or antigen-binding fragment is capable of ting T-cell responses. 49. The method of any one of paragraphs 1, 3-5, 7, 8, 10, 12-19, 21-28, 30, or 32-48, wherein, the population of cells comprises a T-cell. 50. The method of any one of paragraphs 1, 3-5, 7, 8, 10, 12-19, 21-28, 30, or 32-49, wherein the population of cells comprises peripheral blood mononuclear cells. 51. The method of paragraph 50, wherein the peripheral blood mononuclear cells were ed from a human. 52. The method of any one of paragraphs 1, 3-5, 7, 8, 10, 12-19, 21-28, 30, or 32-49, wherein the population of cells is in whole blood. 53. The method of paragraph 52, wherein the whole blood was obtained from a human. 54. The method of any one of paragraphs 1, 3-5, 7, 8, 10, 12-19, 21-28, 30, or 32-49, wherein the population of cells is in an organism. 55. The method of paragraph 54, wherein the population of cells is in a patient having an autoimmune or inflammatory disease. 56. The method of any one of paragraphs 1, 3-5, 7, 8, 10, 12-19, 21-28, 30, or 32-55, n the B- cell is an active B-cell. 57. The method of any one of paragraphs 1, 3-5, 7, 8, 10, 12-19, 21-28, 30, or 32-56, wherein at least about 30% of s in the population of cells are depleted. 58. The method of any one of paragraphs 49-57, wherein less than about 5% of T-cells are depleted. 59. The method of any one of paragraphs 2-4, 6, 7, 9, 11-18, 20-27, 29, or 31-48, further sing administering a second therapeutic agent. 60. The method of paragraph 59, wherein the second therapeutic is ed from the group ting of methotrexate, an anti-CD20 therapeutic, an anti-IL-6 receptor therapeutic, an anti- IL-12/23p40 eutic, a chemotherapeutic, an immunosuppressant, an anti-Interferon beta-1a therapeutic, glatiramer acetate, an anti-α4-integrin therapeutic, imod, an anti-BLys therapeutic, CTLA-Fc, or an anti-TNF therapeutic. 61. The method of paragraph 59, wherein the second eutic is an antibody directed against an antigen selected from a group consisting of CD3, CD14, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD36, CD38, CD40, CD44, CD52, CD55, CD59, CD56, CD70, CD79, CD80, CD103, CD134, CD137, CD138, and CD152. 62. The method of paragraph 59, wherein the second therapeutic is an antibody directed against an antigen selected from the group consisting of IL-2, IL-6, IL-12, IL-23, IL-12/23 p40, IL-17, IFN, TNF, IFN, IL-15, IL-21, IL-1a, IL-1b, IL-18, IL-8, IL-4, GM-CSF, IL-3, and IL-5. 63. The method of any one of 2-4, 6, 7, 9, 11-18, 20-28, 30, 32-47, or 58-62, wherein said autoimmune or inflammatory disease is selected from the group consisting of rheumatoid arthritis, multiple sclerosis, type I diabetes mellitus, idiopathic inflammatory myopathy, systemic lupus erythematosus (SLE), myasthenia gravis, s disease, dermatomyositis, polymyositis, Crohn’s diasease, ulcerative colitis, gastritis, Hashimoto’s thyroiditis, asthma, psoriasis, psoriatic arthritis, dertmatitis, systemic sclerodema and sclerosis, inflammatory bowel disease (IBD), respiratory distress syndrome, meningitis, encephalitis, uveitis, glmerulonephritis, eczema, atherosclerosis, leukocyte adhesion deficiency, Raynaud’s syndrome, Sjögen’s syndrome, Reiter’s disease, Beheet’s diasease, complex nephritis, IgA enphropathy, IgM polyneuropathies, immune-mediated thrombocytopenias, acute thic thrombocytopenic purpura, chronic idiopathic thembocytopenic purpura, tic anemia, myasthenia gravis, lupus nephritis, atopic dermatitis, pemphigus vulgaris, onusmyoclonus syndrome, pure red cell aplasia, mixed cryoglobulinermia, ankylosing spondylitis, hepatitis C-associated cryoglobulinemic vasculitis, chronic focal encephalitis, bullous pemphigoid, hemophilia A, membranoproliferative glomerulonephritis, adult and juvenile omyositis, adult polymyositis, c urticaria, y biliary cirrhosis, neuromyelitis optica, Graves’ roid disease, bullous pemphigoid, membranoproliferative glomerulonephritis, Churg-Strauss syndrome, le onset diabetes, hemolytic , atopic dermatitis, systemic sclerosis, Sjögen’s syndrome and glomerulonephritis, dermatomyositis, ANCA, aplastic anemia, autoimmune hemolytic anemia (AIHA), factor VIII ency, hemophilia A, autoimmune penia, Castleman's syndrome, Goodpasture's syndrome, solid organ transplant rejection, graft versus host disease , autoimmune hepatitis, lymphoid interstitial pneumonitis, HIV, bronchiolitis obliterans (non-transplant), Guillain-Barre Syndrome, large vessel vasculitis, giant cell asu's) arteritis, medium vessel vasculitis, Kawasaki's Disease, polyarteritis nodosa. Wegener’s granulomatosis, microscopic polyangiitis (MPA), Omenn’s syndrome, chronic renal failure, acute infectious mononucleosis,HIV and herpes virus associated diseases.

Still further embodiments are within the scope of the following claims.

Claims (21)

What is claimed

1. An isolated antibody or antigen binding fragment thereof that specifically binds to CD37, wherein said antibody or fragment thereof comprises the heavy chain and light chain variable region CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 171, 172 or 181, and 173 and SEQ ID NOs: 174, 175, and 176, respectively.

2. The isolated antibody or antigen binding fragment thereof of claim 1, wherein said antibody or fragment thereof ses the polypeptide sequences of SEQ ID NO: 177 and SEQ ID NO: 178.

3. The isolated antibody or antigen g fragment thereof of claim 1 or claim 2, wherein said antibody or antigen binding fragment f is murine.

4. The isolated antibody or antigen binding fragment f of any one of claims 1-3, which is a full length antibody.

5. The isolated antibody or antigen binding fragment thereof of any one of claims 1-3, which is an antigen-binding fragment.

6. The isolated dy or n binding fragment thereof of any one of claims 1-5, wherien said antibody or antigen-binding fragment thereof ses a Fab, Fab', F(ab')2, single chain Fv or scFv, disulfide linked Fv, intrabody, IgGACH2, dy, F(ab), tetrabody, triabody, diabody, DVD-Ig, mAb2, (scFv)2, or scFv-Fc.

7. An isolated cell producing the antibody or antigen g fragment thereof of any one of claims 1-6.

8. A method of making an anti-CD37 antibody or antigen-binding fragment thereof comprising (a) culturing the cell of claim 7; and (b) isolating said antibody, or antigen binding nt thereof from said ed cell.

9. An immunoconjugate having the formula (A) - (L) - (C), wherein: (A) is the antibody or antigen binding fragment thereof as defined in any one of claims 1- (L) is a linker; and (C) is a cytotoxic agent; and wherein said linker (L) links (A) to (C).

10. The immunoconjugate of claim 9, wherein said linker is selected from the group consisting of a cleavable , a hydrophilic linker, and a dicarboxylic acid based linker.

11. The immunoconjugate of claim 9, wherein said linker is a non-cleavable linker.

12. The immunoconjugate of any one of claims 9-11, wherein said linker is selected from the group consisting of: N-succinimidyl 4-(2-pyridyldithio)pentanoate (SPP); N-succinimidyl 4-(2-pyridyldithio)butanoate (SPDB) or N-succinimidyl yridyldithio) sulfobutanoate (sulfo-SPDB); N-succinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (SMCC); N-sulfosuccinimidyl 4-(maleimidomethyl) cyclohexanecarboxylate (sulfoSMCC); N-succinimidyl(iodoacetyl)-aminobenzoate (SIAB); and N-succinimidyl-[(N-maleimidopropionamido)-tetraethyleneglycol] ester (NHS-PEG4-maleimide).

13. The immunoconjugate of claim 12, wherein said linker is SMCC or SPDB.

14. The immunoconjugate of any one of claims 9-13, wherein said cytotoxic agent (C) is selected from the group consisting of a maytansinoid, sinoid analog, doxorubicin, a modified doxorubicin, benzodiazepine, taxoid, CC-1065, CC-1065 analog, duocarmycin, duocarmycin , calicheamicin, dolastatin, dolastatin , auristatin, tomaymycin derivative, and leptomycin derivative or a prodrug of the cytotoxic agent.

15. The immunoconjugate of claim 14, wherein said cytotoxic agent (C) is a sinoid.

16. The conjugate of claim 14 or claim 15, wherein said cytotoxic agent (C) is N(2')- deacetyl-N(2')-(3-mercaptooxopropyl)-maytansine (DM1) or N(2')-deacetyl-N(2')-(4- mercaptomethyloxopentyl)-maytansine (DM4).

17. The immunoconjugate of claim 16, wherein said cytotoxic agent (C) is N(2')-deacetyl- N(2')-(3-mercaptooxopropyl)-maytansine (DM1).

18. The immunoconjugate of any one of claims 9-17, wherein the immunoconjugate comprises multiple cytotoxic agents (C) with 3 to 4 (C) per (A).

19. The immunoconjugate of any one of claims 9-18, wherein (L) is SMCC, and n (C) is DM1.

20. The immunoconjugate of any one of claims 9-18, wherein (A) comprises the ptide of SEQ ID NO:177 and the polypeptide of SEQ ID NO:178, wherein (L) is SMCC, and wherein (C) is DM1.

21. An in vitro or ex vivo method for depleting a B-cell comprising ting a population of cells comprising a B-cell with the antibody or antigen binding fragment of any one of claims 1-6 or the immunoconjugate of any one of claims 9-20. ImmunoGen, Inc.