WO2022197699A1 - Human tspan33 antibodies - Google Patents

Abstract

The present disclosure relates to antibodies that specifically bind human TSPAN33, compositions comprising such TSPAN33 antibodies, and methods of using such TSPAN33 antibodies.

Description

HUMAN TSPAN33 ANTIBODIES

CROSS REFERENCE TO RELATED APPLICATIONS

The present application claims the benefit of priority to U.S. Provisional Patent Application No. 63/162,197, filed on March 17, 2021, the entire disclosure of which is incorporated by reference herein.

SEQUENCE LISTING

A Sequence Listing accompanies this application and is submitted as an ASCII text file of the sequence listing named “083389_01622_X30022_SequenceListing.txt” which is 95,709 bytes in size and was created on March 15, 2022. The sequence listing is electronically submitted via EFS-Web with the application and is incorporated herein by reference in its entirety.

The present disclosure is in the field of medicine. Particularly the present disclosure relates to antibodies that specifically bind human TSPAN33, compositions comprising such TSPAN33 antibodies, and methods of using such TSPAN33 antibodies.

BACKGROUND

B cells can produce antibodies against various antigens and thus play an important role in the humoral immune response. Additionally, B cells also function as antigen presenting cells (APC) and secrete cytokines. B cells express B cell receptor (BCR) on their cell membranes; and BCR allows the B cell to bind a specific antigen, against which it will initiate an antibody response. Dysregulation of B cells is associated with a variety of disorders.

Human TSPAN33 (also known as Tetraspanin-33, Penumbra (hPen), Proerythroblast new membrane, BAAM antigen) is a member of the transmembrane 4 superfamily (TM4SF) of proteins that associate with other molecules such as integrins or other tetraspanins to regulate the function of the associated molecules mediating diverse biological processes, including cell activation, survival, proliferation, adhesion, and migration. Human TSPAN33 has been found to associate with other Tetraspanin family members as well as with a repertoire of other transmembrane and intracellular molecules, playing an active role during B-lymphocyte cytoskeleton and plasma membrane-related phenomena, including protrusion formation, adhesion, activation, phagocytosis, and cell motility including metastasis. TSPAN33 has been reported as being expressed by erythroid cells, kidney cells and activated B cells (Luu et al., Clin Immunol., 2013; 149:388-99). Human TSPAN33 has been reported as being upregulated in certain cancers, and as being constitutively expressed in some B cell lymphomas. (Id.; Perez-Martinez, et al., Scandinavian J. of Immunol., 2017; 86(1): 23-30).

Antibodies to B-cell receptors such as CD 19 and CD20, which are expressed on activated and resting human B cells, are known and approved for therapeutic use or are in clinical development for treatment of B cell associated disorders. However, to date, no known antibody that specifically binds human TSPAN33 has been approved for therapeutic use, nor is there an antibody that specifically binds human TSPAN33 in clinical development. Therefore, there remains a need for antibodies that specifically bind human TSPAN33 for use in treatment of B cell associated disorders, particularly disorders associated with activated B cells.

DETAILED DESCRIPTION

The present disclosure provides antibodies that specifically bind human TSPAN33 and inhibit B cell responses (e.g., proliferation, adhesion, activation, motility, survival, and / or migration), compositions comprising such TSPAN33 antibodies, and methods of using such TSPAN33 antibodies. Particularly the present disclosure provides human TSPAN33 antibodies that specifically deplete activated B cells and / or that specifically target disorders associated with activated B cells. Such human TSPAN33 antibodies can be used to treat B cell associated disorders, such as autoimmune diseases, particularly those associated with activated B cells, without destroying important immune cells including resting B cells, and thus avoid problematic concurrent immunocompromise, long-term immune suppression, and other complications resulting from targeting resting B cells. Such human TSPAN33 antibodies can be further used to treat B cell associated cancer, for example, B cell lymphomas or cancers expressing TSPAN33. The human TSPAN33 antibodies provided herein have one or more of the following properties: 1) bind human TSPAN33 with desirable binding affinities and / or association and dissociation rates, 2) bind human TSPAN33 on activated human B cells and inhibit proliferation, adhesion, activation, motility, survival and / or differentiation, 3) deplete activated B cells, 4) induce effector function mediated killing, 5) do not significantly bind resting B cells, and / or 6) low immunogenicity risk.

In some embodiments, the antibodies of the present disclosure that specifically bind human TSPAN33 are fully human antibodies. In some embodiments, the antibodies of the present disclosure specifically bind human TSPAN33 on activated B cells. In further embodiments, antibodies of the present disclosure do not deplete resting (non-activated) B cells. In further embodiments, the antibodies of the present disclosure bind human TSPAN33 on activated B cells and deplete the activated B cells. In yet further embodiments, the antibodies of the present disclosure induce effector function mediated killing.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, wherein the antibody or antibody binding fragment thereof comprise a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDR) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDR) LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 1, the HCDR2 comprises SEQ ID NO: 2, the HCDR3 comprises SEQ ID NO: 3, the LCDR1 comprises SEQ ID NO: 4, the LCDR2 comprises SEQ ID NO: 5, and the LCDR3 comprises SEQ ID NO: 6. In some embodiments, the human TSPAN33 antibodies comprise a VH comprising SEQ ID NO: 7 and a VL comprising SEQ ID NO: 8. In some embodiments, the antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, comprises a heavy chain (HC) comprising SEQ ID NO: 9 and a light chain (LC) comprising SEQ ID NO: 10.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, and comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 13, the HCDR2 comprises SEQ ID NO: 14, the HCDR3 comprises SEQ ID NO: 15, the LCDR1 comprises SEQ ID NO: 16, the LCDR2 comprises SEQ ID NO: 17, and the LCDR3 comprises SEQ ID NO: 18. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 19 and a VL comprising SEQ ID NO: 20. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 21 and a LC comprising SEQ ID NO: 22.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, and comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 25, the HCDR2 comprises SEQ ID NO: 26, the HCDR3 comprises SEQ ID NO: 27, the LCDR1 comprises SEQ ID NO: 16, the LCDR2 comprises SEQ ID NO: 28, and the LCDR3 comprises SEQ ID NO: 29. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 30 and a VL comprising SEQ ID NO: 31. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 32 and a LC comprising SEQ ID NO: 33.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, and comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 36, the HCDR2 comprises SEQ ID NO: 37, the HCDR3 comprises SEQ ID NO: 38, the LCDR1 comprises SEQ ID NO: 39, the LCDR2 comprises SEQ ID NO: 40, and the LCDR3 comprises SEQ ID NO: 41. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 42 and a VL comprising SEQ ID NO: 43. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 44 and a LC comprising SEQ ID NO: 45. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 74 and a VL comprising SEQ ID NO: 75. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 76 and a LC comprising SEQ ID NO: 77.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, and comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 36, the HCDR2 comprises SEQ ID NO: 37, the HCDR3 comprises SEQ ID NO: 48, the LCDR1 comprises SEQ ID NO: 39, the LCDR2 comprises SEQ ID NO: 40, and the LCDR3 comprises SEQ ID NO: 49. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 50 and a VL comprising SEQ ID NO: 51. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 52 and a LC comprising SEQ ID NO: 53.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, and comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 56, the HCDR2 comprises SEQ ID NO: 57, the HCDR3 comprises SEQ ID NO: 58, the LCDR1 comprises SEQ ID NO: 59, the LCDR2 comprises SEQ ID NO: 60, and the LCDR3 comprises SEQ ID NO: 61. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 62 and a VL comprising SEQ ID NO: 63. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 64 and a LC comprising SEQ ID NO: 65.

In some embodiments, the present disclosure provides an antibody or antibody binding fragment thereof, that specifically binds human TSPAN33, and comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein the HCDR1 comprises SEQ ID NO: 56, the HCDR2 comprises SEQ ID NO: 57, the HCDR3 comprises SEQ ID NO: 58, the LCDR1 comprises SEQ ID NO: 59, the LCDR2 comprises SEQ ID NO: 60, and the LCDR3 comprises SEQ ID NO: 68. In some embodiments, the antibody or antibody binding fragment thereof that specifically bind TSPAN33 comprise a VH comprising SEQ ID NO: 62 and a VL comprising SEQ ID NO: 69. In some embodiments, the antibody comprises a HC comprising SEQ ID NO: 70 and a LC comprising SEQ ID NO: 71.

In some embodiments of the present disclosure, the antibody specifically binding human TSPAN33 has a human IgGl isotype. In some embodiments, the antibody specifically binding human TSPAN33 has a modified human IgGl Fc region which enhances the IgGl Fc effector function of the antibody, such as antibody-dependent cell cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC). In further embodiments, the enhanced effector function of the modified antibodies enhances B cell depletion. In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising one or more of the following amino acid substitutions compared to the wild-type human IgGl Fc region: P247I, I332E, A339Q (all positions numbered according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising a P247I substitution (according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising a I332E substitution (according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising a A339Q substitution (according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising P247I and I332E substitutions (all positions numbered according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising P247I and A339Q substitutions (all positions numbered according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising I332E and A339Q substitutions (all positions numbered according to the EU Index Numbering). In some embodiments, the human TSPAN33 antibody has a modified human IgGl Fc region comprising P247I, I332E, and A339Q substitutions (all positions numbered according to the EU Index Numbering). In some embodiments of the present disclosure, the human TSPAN33 antibody has a human IgG4 isotype. In some embodiments, the human TSPAN33 antibody has a modified human IgG4 Fc Region.

In some embodiments, the antibody specifically binding human TSPAN33 has a modified human IgGl framework region. In some embodiments, the modifications are in the framework region of VH. In some embodiments, the modifications are in the framework region of VL. In some embodiments, the modifications are in the framework regions of VH and the VL. In further embodiments, the modified human IgGl framework region lowers the immunogenicity risk of the antibody.

Some embodiments of the present disclosure provide nucleic acids encoding a heavy chain or light chain, or a VH or VL, of the novel antibodies that specifically bind human TSPAN33, or vectors comprising such nucleic acids .

In some embodiments, the present disclosure provides a nucleic acid comprising a sequence of SEQ ID NO: 11, 12, 23, 24, 34, 35, 46, 47, 54, 55, 66, 67, 72, 73, 78, or 79 . In some embodiments, nucleic acids encoding a heavy chain or light chain of the antibodies specifically binding human TSPAN33 are provided. In some embodiments nucleic acids comprising a sequence encoding SEQ ID NO: 9, 10, 21, 22, 32, 33, 44, 45, 52, 53, 64, 65, 70, 71, 76, or 77 are provided. In some embodiments, nucleic acids comprising a sequence encoding an antibody heavy chain that comprises SEQ ID NO: 9, 21, 32, 44, 52, 64, 70, or 76 are provided. For example, the nucleic acid can comprise a sequence selected from SEQ ID NO: 11, 23, 34, 46, 54, 66, 72, or 78. In some embodiments, nucleic acids comprising a sequence encoding an antibody light chain that comprises SEQ ID NO: 10, 22, 33, 45, 53, 65, 71, or 77 is provided. For example, the nucleic acid can comprise a sequence selected from SEQ ID NO: 12, 24, 35, 47, 55, 67, 73, or 79.

In some embodiments of the present disclosure, nucleic acids encoding a VH or VL of the antibodies specifically binding human TSPAN33 are provided. In some embodiments, nucleic acids comprising a sequence encoding SEQ ID NO: 7, 8, 19, 20, 30, 31, 42, 43, 50, 51, 62, 63, 69, 74, or 75 are provided. In some embodiments, nucleic acids comprising a sequence encoding an antibody VH that comprises SEQ ID NO: 7, 19, 30, 42, 50, 62, or 74 are provided. In some embodiments, nucleic acids comprising a sequence encoding an antibody VL that comprises SEQ ID NO: 8, 20, 31, 43, 51, 63, 69, or 75 is provided.

Some embodiments of the present disclosure provide vectors comprising a nucleic acid sequence encoding an antibody heavy chain or light chain. For example, such vectors can comprise a nucleic acid sequence encoding SEQ ID NO: 9, 21, 32, 44, 52, 64, 70, or 76. In some embodiments, the vector comprises a nucleic acid sequence encoding SEQ ID NO: 10, 22, 33, 45, 53, 65, 71, or 77.

Provided herein are also vectors comprising a nucleic acid sequence encoding an antibody VH or VL. For example, such vectors can comprise a nucleic acid sequence encoding SEQ ID NO: 7, 19, 30, 42, 50, 62, or 74. In some embodiments, the vector comprises SEQ ID NO: 8, 20, 31, 43, 51, 63, 69, or 75.

Provided herein are also vectors comprising a first nucleic acid sequence encoding an antibody heavy chain and a second nucleic acid sequence encoding an antibody light chain. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 9, 21, 32, 44, 52, 64, 70, or 76, and a second nucleic acid sequence encoding SEQ ID NO: 10, 22, 33, 45, 53, 65, 71, or 77.

In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 9 and a second nucleic acid sequence encoding SEQ ID NO: 10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 21 and a second nucleic acid sequence encoding SEQ ID NO: 22. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 32 and a second nucleic acid sequence encoding SEQ ID NO: 33. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 44 and a second nucleic acid sequence encoding SEQ ID NO: 45. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 52 and a second nucleic acid sequence encoding SEQ ID NO: 53. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 64 and a second nucleic acid sequence encoding SEQ ID NO: 65. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 70 and a second nucleic acid sequence encoding SEQ ID NO: 71. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 76 and a second nucleic acid sequence encoding SEQ ID NO: 77.

Also provided are compositions comprising a first vector comprising a nucleic acid sequence encoding an antibody heavy chain, and a second vector comprising a nucleic acid sequence encoding an antibody light chain. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, 21,

32, 44, 52, 64, 70, or 76, and a second nucleic acid sequence encoding SEQ ID NO: 10, 22,

33, 45, 53, 65, 71, or 77.

In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 21 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 22. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 32 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 33. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 44 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 45. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 52 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 53. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 64 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 65. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 70 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 71. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 76 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 77.

Nucleic acids of the present disclosure may be expressed in a host cell, for example, after the nucleic acids have been operably linked to an expression control sequence. Expression control sequences capable of expression of nucleic acids to which they are operably linked are well known in the art. An expression vector may include a sequence that encodes one or more signal peptides that facilitate secretion of the polypeptide(s) from a host cell. Expression vectors containing a nucleic acid of interest (e.g., a nucleic acid encoding a heavy chain or light chain of an antibody) may be transferred into a host cell by well-known methods, e.g., stable or transient transfection, transformation, transduction or infection. Additionally, expression vectors may contain one or more selection markers, e.g., tetracycline, neomycin, and dihydrofolate reductase, to aide in detection of host cells transformed with the desired nucleic acid sequences.

In another aspect, provided herein are cells, e.g., host cells, comprising the nucleic acids, vectors, or nucleic acid compositions described herein. A host cell may be a cell stably or transiently transfected, transformed, transduced or infected with one or more expression vectors expressing all or a portion of an antibody described herein. In some embodiments, a host cell may be stably or transiently transfected, transformed, transduced or infected with an expression vector expressing HC and LC polypeptides of an antibody of the present disclosure. In some embodiments, a host cell may be stably or transiently transfected, transformed, transduced or infected with a first vector expressing HC polypeptides and a second vector expressing LC polypeptides of an antibody described herein. Such host cells, e.g., mammalian host cells, can express the antibodies that specifically bind human TSPAN33 as described herein. Mammalian host cells known to be capable of expressing antibodies include CHO cells, HEK293 cells, COS cells, andNSO cells.

In some embodiments, the cell, e.g., host cell, comprises a vector comprising a first nucleic acid sequence encoding SEQ ID NO: 9, 21, 32, 44, 52, 64, 70, or 76 and a second nucleic acid sequence encoding SEQ ID NO: 10, 22, 33, 45, 53, 65, 71, or 77.

In some embodiments, the cell, e.g., host cell, comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, 21, 32, 44, 52, 64, 70, or 76 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10, 22, 33, 45, 53, 65, 71, or 77.

The present disclosure further provides a process for producing an antibody or antibody binding fragments thereof that specifically binds human TSPAN33 described herein by culturing the host cell described above, e.g., a mammalian host cell, under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium. The culture medium, into which an antibody has been secreted, may be purified by conventional techniques. Various methods of protein purification may be employed, and such methods are known in the art and described, for example, in Deutscher, Methods in Enzymology 182: 83-89 (1990) and Scopes, Protein Purification: Principles and Practice, 3rd Edition, Springer, NY (1994).

The present disclosure further provides antibodies or antibody binding fragments thereof produced by any of the processes described herein.

In another aspect, provided herein are pharmaceutical compositions comprising an antibody, nucleic acid, or vector described herein. Such pharmaceutical compositions can also comprise one or more pharmaceutically acceptable excipient, diluent or carrier. Pharmaceutical compositions can be prepared by methods well known in the art (e.g., Remington: The Science and Practice of Pharmacy , 22nd ed. (2012), A. Loyd et ah, Pharmaceutical Press).

The antibodies or antibody binding fragments thereof that specifically bind human TSPAN33, nucleic acids, vectors, or pharmaceutical compositions described herein can be used for treating B cell associated disorders. Because of their critical role in regulating the immune system, dysregulation of B cells is associated with a variety of disorders. B cell associated disorders include autoimmune diseases, which are caused by activation of self reactive B cells and T cells, and lymphomas and leukemias, which are caused by excessive and/or uncontrolled B cell proliferation. Such autoimmune diseases include, but are not limited to systemic lupus erythematosus, granulomatosis with polyangiitis (GPA) (Wegener’s Granulomatosis), microscopic polyangiitis (MPA), pemphigus vulgaris (PV), Lupus nephritis, multiple sclerosis, rheumatoid arthritis, Sjogren’s syndrome, myasthenia gravis. In some embodiments, the B cell associated disorder is characterized by activated B cells. In some embodiments, the B cell associated disorder is cancer, e.g., B cell lymphomas, such as Non-Hodgkin’s Lymphoma (NHL), Burkitt lymphoma, diffuse large B-cell lymphoma; other cancers expressing TSPAN33, including, but not limited to, Chronic Lymphocytic Leukemia (CLL).

In some embodiments, the antibodies or antibody binding fragments thereof described herein bind human TSPAN33 expressed on activated B cells, and offer advantages over other B cell binding antibodies that bind both activated and/or resting B cells for treating B cell associated diseases such as autoimmune diseases and cancer. Such advantages include avoidance of problematic concurrent immunocompromise, long-term immune suppression, compromised vaccination and other complications resulting from binding and / or depleting resting B cells.

In some embodiments, provided herein are methods of treating a B cell associated disorder, e.g., an autoimmune disease or cancer, in a subject (e.g., a human patient) in need thereof, by administering to the subject a therapeutically effective amount of an antibody or antibody binding fragment thereof that specifically binds human TSPAN33, a nucleic acid encoding such an antibody that specifically binds human TSPAN33, a vector comprising such a nucleic acid, or a pharmaceutical composition comprising such an antibody that specifically binds human TSPAN33, nucleic acid or vector, as described herein. The antibodies, nucleic acids, vectors, or pharmaceutical compositions described herein may be administered by parenteral routes (e.g., subcutaneous and intravenous). In some embodiments, the B cell associated disorder is an autoimmune disease. Such an autoimmune disease includes, but is not limited to, systemic lupus erythematosus, granulomatosis with polyangiitis (GPA) (Wegener’s Granulomatosis), microscopic polyangiitis (MPA), pemphigus vulgaris (PV), lupus nephritis, multiple sclerosis, rheumatoid arthritis, Sjogren’s syndrome, myasthenia gravis. In some embodiments, the B cell associated disorders is characterized by activated B cells. In some embodiments, the B cell associated disorder is cancer. Examples of cancer include, but are not limited to, B cell lymphomas, such as to Non-Hodgkin’s Lymphoma (NHL), Burkitt lymphoma, diffuse large B-cell lymphoma; other cancers expressing TSPAN33, including but not limited to Chronic Lymphocytic Leukemia (CLL). Such methods can further include administering one or more chemotherapeutic agents to the subject. In some embodiments, the chemotherapeutic agent comprises fludarabine or cyclophosphamide. In some embodiments, the chemotherapeutic agent is administered in simultaneous, separate, or sequential combination with an antibody or antibody binding fragment thereof that specifically binds human TSPAN33 described herein or pharmaceutical composition thereof. Embodiments of the present disclosure further provide methods of treating cancer with an antibody or antibody binding fragment thereof that specifically binds human TSPAN33 or pharmaceutical composition thereof, in simultaneous, separate, or sequential combination with ionizing radiation.

Also provided herein are, antibodies or antibody binding fragments thereof that specifically bind human TSPAN33, nucleic acids, vectors, or pharmaceutical compositions described herein for use in therapy. Furthermore, the present disclosure also provides, antibodies or antibody binding fragments thereof that specifically bind human TSPAN33, nucleic acids, vectors, or pharmaceutical compositions described herein for use in the treatment of a B cell associated disorder, e.g., an autoimmune disease or cancer. In some embodiments, the B cell associated disorder is autoimmune disease. Such autoimmune diseases include, but are not limited to, systemic lupus erythematosus, granulomatosis with polyangiitis (GPA) (Wegener’s Granulomatosis), microscopic polyangiitis (MPA), pemphigus vulgaris (PV), lupus nephritis, multiple sclerosis, rheumatoid arthritis, Sjogren’s syndrome, myasthenia gravis. In some embodiments, the B cell associated disorders is characterized by activated B cells. In some embodiments, the B cell associated disorder is cancer. Examples of cancer include, but are not limited to, B cell lymphomas, such as to Non-Hodgkin’s Lymphoma (NHL), Burkitt lymphoma, diffuse large B-cell lymphoma; other cancers expressing TSPAN33, including but not limited to Chronic Lymphocytic Leukemia (CLL).

Provided herein are use of the antibodies or antibody binding fragments thereof that specifically bind human TSPAN33, nucleic acids, vectors, or pharmaceutical compositions described herein in the manufacture of a medicament for the treatment of a B cell associated disorder, e.g., an autoimmune disease or cancer. In some embodiments, the B cell associated disorder is autoimmune disease. Such autoimmune disease includes, but is not limited to systemic lupus erythematosus, granulomatosis with polyangiitis (GPA) (Wegener’s Granulomatosis), microscopic polyangiitis (MPA), pemphigus vulgaris (PV), Lupus nephritis, multiple sclerosis, rheumatoid arthritis, Sjogren’s syndrome, myasthenia gravis. In some embodiments the B cell associated disorders is characterized by activated B cells. In some embodiments, the B cell associated disorder is cancer. Examples of cancer include but are not limited to B cell lymphomas, such as to Non-Hodgkin’s Lymphoma (NHL), Burkitt lymphoma, diffuse large B-cell lymphoma; other cancers expressing TSPAN33, including but not limited to Chronic Lymphocytic Leukemia (CLL).

“Chemotherapeutic agent” or “chemotherapeutic” as used interchangeably herein, is a chemical agent or drug that is selectively destructive to cancer cells and tissues. Chemotherapeutic agent may include but is not limited to compounds such as, taxane compounds, compounds that act via taxane mechanisms, platinum compounds, anthracycline compounds, antimetabolites, alkylating agents, epipodophyllotoxin compounds, camptothecin compounds, or any combination thereof. Chemotherapeutic agents can be administered alone or in combination with other therapeutic agents. In some embodiments, a chemotherapeutic agent comprises fludarabine or cyclophosphamide.

“Ionizing radiation” is radiation of certain wavelengths used to destroy or damage cancer cells. Ionizing radiation includes radon, x-rays, gamma rays, and other forms of high-energy radiation. Ionizing radiation may include external radiation (or external beam radiation), internal radiation (or brachytherapy) or systemic radiation.

As used herein, the term “a,” “an,” “the” and similar terms used in the context of the present disclosure (especially in the context of the claims) are to be construed to cover both the singular and plural unless otherwise indicated herein or clearly contradicted by the context.

The term “antibody,” as used herein, refers to an immunoglobulin molecule that binds an antigen. Embodiments of an antibody include a monoclonal antibody, polyclonal antibody, human antibody, humanized antibody, chimeric antibody, or conjugated antibody. The antibodies can be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any subclass (e.g., IgGl, IgG2, IgG3, IgG4). An exemplary antibody is an immunoglobulin G (IgG) type antibody comprised of four polypeptide chains: two heavy chains (HC) and two light chains (LC) that are cross- linked via inter-chain disulfide bonds. The amino-terminal portion of each of the four polypeptide chains includes a variable region of about 100-125 or more amino acids primarily responsible for antigen recognition. The carboxyl-terminal portion of each of the four polypeptide chains contains a constant region primarily responsible for effector function. Each heavy chain is comprised of a heavy chain variable region (VH) and a heavy chain constant region. Each light chain is comprised of a light chain variable region (VL) and a light chain constant region. The IgG isotype may be further divided into subclasses (e.g., IgGl, IgG2, IgG3, and IgG4).

The VH and VL regions can be further subdivided into regions of hyper-variability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). The CDRs are exposed on the surface of the protein and are important regions of the antibody for antigen binding specificity. Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Herein, the three CDRs of the heavy chain are referred to as “HCDRl, HCDR2, and HCDR3” and the three CDRs of the light chain are referred to as “LCDR1, LCDR2 and LCDR3”. The CDRs contain most of the residues that form specific interactions with the antigen. Assignment of amino acid residues to the CDRs may be done according to the well-known schemes, including those described in Rabat (Rabat et ak, “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md. (1991)), Chothia (Chothia et ak, “Canonical structures for the hypervariable regions of immunoglobulins”, Journal of Molecular Biology, 196, 901-917 (1987); Al-Lazikani et ak, “Standard conformations for the canonical structures of immunoglobulins”, Journal of Molecular Biology, 273, 927-948 (1997)), North (North et al., “A New Clustering of Antibody CDR Loop Conformations”, Journal of Molecular Biology, 406, 228-256 (2011)), or IMGT (the international ImMunoGeneTics database available on at www.imgt.org; see Lefranc et ak, Nucleic Acids Res. 1999; 27:209-212). The North CDR definitions are used for the antibodies that specifically bind human TSPAN33 described herein. Exemplary embodiments of antibodies of the present disclosure also include antibody fragments or antigen-binding fragments, which comprise at least a portion of an antibody retaining the ability to specifically interact with an antigen such as Fab, Fab’, F(ab’)2, Fv fragments, scFv, scFab, disulfide-linked Fvs (sdFv), a Fd fragment and linear antibodies.

The terms “bind” and “binds” as used herein are intended to mean, unless indicated otherwise, the ability of a protein or molecule to form a chemical bond or attractive interaction with another protein or molecule, which results in proximity of the two proteins or molecules as determined by common methods known in the art.

The term “TSPAN33” as used herein, unless stated otherwise, refers to any native, mature TSPAN33 that results from processing of a TSPAN33 precursor protein in a cell. The term includes TSPAN33 from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus or rhesus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring variants of TSPAN33, e.g., splice variants or allelic variants. The amino acid sequence of an example of human TSPAN33 is known in the art, e.g., NCBI reference sequence XP_006716023.1 (SEQ ID NO: 80). The term “TSPAN33” is used herein to refer collectively to all known human TSPAN33 isoforms and polymorphic forms.

The term “activated B cells” refers to non-resting B cells. B cell activation is triggered by binding of specific antigens to the B cell receptor (BCR). Various B cell associated disorders have been identified as having an activated B cell phenotype, and TSPAN33 has been reported as being expressed on activated B cells and not on resting B cells (Luu et ak, Clin Immunol., 2013; 149:388-99).

The term “Fc region” as used herein refers to a region of an antibody, which comprises the CH2 and CH3 domains of the antibody heavy chain. Optionally, the Fc region may include a portion of the hinge region or the entire hinge region of the antibody heavy chain. Biological activities such as effector function are attributable to the Fc region, which vary with the antibody isotype. Examples of antibody effector functions include, Fc receptor binding, antibody-dependent cell mediated cytotoxicity (ADCC), antibody- dependent cell mediated phagocytosis (ADCP), Clq binding, complement dependent cytotoxicity (CDC), phagocytosis, down regulation of cell surface receptors (e.g. B cell receptor); and B cell activation. The terms “nucleic acid” or “polynucleotide”, as used interchangeably herein, refer to polymers of nucleotides, including single-stranded and / or double-stranded nucleotide- containing molecules, such as DNA, cDNA and RNA molecules, incorporating native, modified, and / or analogs of, nucleotides. Polynucleotides of the present disclosure may also include substrates incorporated therein, for example, by DNA or RNA polymerase or a synthetic reaction.

The term “subject”, as used herein, refers to a mammal, including, but are not limited to, a human, chimpanzee, ape, monkey, cattle, horse, sheep, goat, swine, rabbit, dog, cat, rat, mouse, guinea pig, and the like. Preferably, the subject is a human.

The term “therapeutically effective amount,” as used herein, refers to an amount of a protein or nucleic acid or vector or composition that will elicit the biological or medical response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc. In a non-limiting embodiment, the term “a therapeutically effective amount” refers to the amount of a protein or nucleic acid or vector or composition that, when administered to a subject, is effective to at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease.

As used herein, “treatment” or “treating” refers to all processes wherein there may be a slowing, controlling, delaying or stopping of the progression of the disorders or disease disclosed herein, or ameliorating disorder or disease symptoms, but does not necessarily indicate a total elimination of all disorder or disease symptoms. Treatment includes administration of a protein or nucleic acid or vector or composition for treatment of a disease or condition in a patient, particularly in a human.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows the binding specificity of human TSPAN33 antibodies to human TSPAN33 in an antibody down ELISA assay format.

FIG. 2 shows the binding specificity of human TSPAN33 antibodies to human TSPAN33 in a capture ELISA assay format.

FIGs. 3A-3F show the binding specificity of human TSPAN33 antibodies to CHO cells expressing TSPAN33.

FIG. 4 shows the binding specificity of human TSPAN33 antibodies to Daudi cells. FIG. 5 shows the binding specificity of human TSPAN33 antibodies to activated B cells. FIGs. 6A-6C show the human TSPAN33 antibodies induce ADCC activity on unactivated Daudi cells.

FIGs. 7A-7B show the human TSPAN33 antibodies induce ADCC activity on activated Daudi cells.

FIG. 8 shows the human TSPAN33 antibodies may induce CDC activity on CHO Cells expressing TSPAN33.

EXAMPLES

The following examples are offered to illustrate, but not to limit, the claimed invention.

Example 1. Generation of human TSPAN33 antibodies

To develop antibodies specific to human TSPAN33, transgenic mice with human immunoglobulin variable regions are immunized with His-tagged human TSPAN33 large extracellular loop (LEL) fused to monovalent human Fc. Mouse B cells positive for human TSPAN33 binding are sorted by fluorescence-activated cell sorting (FACS) and antibodies are cloned by single cell polymerase chain reaction (PCR). A panel of antibodies are cloned and expressed and screened by ELISA for binding to human TSPAN33 LEL VHH fusion. Positive clones are subsequently screened for binding to TSPAN33 expressed on CHO cells by FACS.

The antibodies of the invention can be synthesized and purified by well-known methods. An appropriate host cell, such as Chinese hamster ovarian cells (CHO), can be either transiently or stably transfected with an expression system for secreting antibodies using a predetermined HC:LC vector ratio if two vectors are used, or a single vector system encoding both heavy chain and light chain. Clarified media, into which the antibody has been secreted, can be purified using the commonly used techniques.

Example 2. Binding Activity of the human TSPAN33 Antibodies

Binding affinity and kinetics: The binding affinity and binding kinetics of exemplified human TSPAN33 antibodies to the TSPAN33-LEL VHH fusion protein is determined by Surface plasmon resonance (SPR) using Biacore™ 8K (GE Healthcare). The procedure generally follows the “Instrument Handbook”. Briefly, polyclonal goat anti-human IgG- Fc (Jackson Immunology cat#109005098) is immobilized onto Series S CM5 sensor chips (GE Healthcare Cat.: #29104988) using standard amine coupling

(NHS/EDC/ethanolamine) protocols provided by the manufacturer’s protocols and a 10 mM Acetate pH 4.5 binding buffer/HBS-EP running buffer. Immobilization levels are between 10-12K resonance units (RUs). The antibody is first captured on Biacore™ goat anti-human IgG-Fc Chip followed by flowing a serial concentration of TSPAN33-LEL VHH fusion protein from 1000 nM down to 3.91 nM in 2-fold serial dilution in PBS-P20- BSA (0.005% surfactant P20, O.lmg/mL BSA).

All binding kinetic experiments are performed at 37 °C. Multi-cycle kinetics setting that runs each analyte concentration in a separate cycle regenerating the surface after each sample injection is used. To measure binding kinetics, the test antibodies are diluted to 10 pg/mL in PBS pH 7.4 and captured onto these chip surfaces to a level of 49-63 RU by injection at 10 pL/min using an HBS-EP running buffer. Next, TSPAN33-LEL fusion protein is diluted into HBS-EP at 1000, 500, 250, 125, 62.5, 31.25, 15.625, 7.8125, 3.91 and 0 nM and injected (180 s at 50 pL/min) in succession over the sensor chip surfaces containing the captured mAh variants. Dissociation of the antigen is allowed to occur for 600 s. Between each antigen injection, mAh and non-dissociated antigen are removed from the sensor chip surface with injection of 10 mM glycine, pH 1.5, buffer (regeneration). The regeneration is optimized to maintain consistent surface properties from cycle to cycle. The data is fit to a 1:1 binding model to derive ka and kd, and to calculate KD (Table Below). The results as demonstrated in Table 1, show exemplary SPR data of the exemplified human TSPAN33 antibodies.

Table 1. Exemplary Binding Affinities and kinetics of human TSPAN33 antibodies

Antigen Down Elisa Binding. Binding affinity of the exemplified human TSPAN33 antibodies to human TSPAN33 fusion protein are tested in an ELISA assay. A 96 well microtiter plate is coated with the recombinant human TSPAN33-LEL fused to a human VHH domain at 2 pg/mL(50 pL/well) in phosphate buffered saline (PBS) overnight. Next day, plate is washed and blocked for lhr at room temperature with 100 pL/well of casein buffer. The plate is washed three times with PBST (PBS with 0.1% Tween) and the TSPAN33 antibodies (4D4, 11E8, S3, and S24) and IgGl negative control antibody (Neg IgGl) are serially diluted 3 -fold from 2 pg/mL in blocking buffer and added to the TSPAN33-LEL fusion protein coated plate at 50 uL/well, and incubated for 1 hour at 37 °C. The plate is washed three times with PBST and anti -human kappa alkaline phosphatase diluted 1 : 1000 in PBS Is added at 50 ul/well and incubated for 1 hour at 37 °C. The plate is washed three times with PBST, and 50 pL of AMP-PMP substrate diluted 1:30 in deionized water is added. Absorbance is read at 560 nm on a Spectramax (Molecular Devices) plate reader.

Capture Elisa Binding: Binding affinity of the exemplified human TSPAN33 antibodies to human TSPAN33-LEL VHH fusion protein are tested in an ELISA assay. 96 well microtiter plates are coated overnight at 4 °C at 2 pg/mL (50 pL/well) of goat anti-human Fc specific polyclonal antibody in PBS. Following overnight incubation, plates are washed and blocked with 100 pL of casein buffer for 1 hour at room temperature. Plates are then washed three times using PBST (PBS with 0.1% Tween) and TSPAN33 antibodies (4D4, 11E8, S3, and S24) and an IgGl negative control antibody (Neg IgGl) diluted at 2 pg/mL in PBS casein are added at 50 pL/well and incubated for 1 hour at 37 °C. Plates are washed three times using PBST and biotinylated TSPAN33-LEL VHH-fusion protein is serially diluted 3-fold from 10 pg/mL and added at 50 pL/well to the plates, and plates are further incubated for 1 hour at 37 °C. Plates are washed three times using PBST, and 200 pL/well of PBS casein is added and incubated for 2 hours at 37 °C to allow for differential dissociation of the antibodies. The plates are then washed, and 50 pL of neutravidin alkaline phosphatase-conjugate diluted 1 : 1000 in PBS casein is added and incubated for 30 min at 37 °C. The plates are washed three times and 50 pL of AMP-PMP substrate diluted 1:30 in deionized water is added and the absorbance is read at 560 nm on a Spectramax (Molecular Devices) plate reader. The results as demonstrated in Figure 1 and Figure 2, show that human TSPAN33 antibodies 4D4, 11E8, S3, and S24 specifically bind the recombinant human TSPAN33- LEL VHH fusion protein.

Cell surface binding: Binding of the exemplified human TSPAN33 antibodies to CHO cells stably expressing TSPAN33 and Daudi B lymphoblast cells is tested in a Fluorescence Activated Cell Sorting (FACS) assay. CHO cells stably expressing different membrane- bound human tetraspanin family members (TSPAN33, TSPAN5, TSPAN14, TSPAN17) and Daudi cells are added to 96-well cell culture plates (Falcon 353077) at 1 xlO⁵ cells/well in FACS buffer (PBS, 2.5% FBS, 20 mM HEPES, 2 mM EDTA). Cells are placed on ice for 10 minutes, and 50 pL of human TSPAN33 antibodies (11E8, S24, and S3) and an IgGl isotype control antibody are added to the wells at a final concentration of 1 pg/ml, and incubated for 1 hour at 4 °C. Cells are then washed 3 times by centrifuging for 5 min at 1000 RPM at 4 °C, and resuspended in 100 pL secondary antibody (PE-labelled goat anti human IgG Fc (Jackson ImmunoResearch cat#109-l 15-098)) diluted 1:800 in FACS buffer, and incubated for 1 hour at 4 °C. For positive controls, commercially available antibodies from BioLegend^® and R&D are used, along with secondary antibody Alexa 488 labelled goat anti-mouse IgG,Fc (Jackson ImmunoResearch cat#l 15-545-071). Cells are then washed as previously described and then resuspended in 100 pL FACS buffer with sytox blue (Thermofisher S34857) diluted at a 1:10,000. Samples are analyzed by flow cytometry on a Propel Labs’ ZE5 cytometer using Everest software. Analysis is performed using FlowJo vlO.7.1, and median fluorescent intensities (MFI) are reported for either PE or Alexa Fluor 488.

The results as demonstrated in Table 2, and Figures 3 A-3F, and Figure 4 show that the human TSPAN33 antibodies 11E8, S24, and S3 specifically recognize and bind human TSPAN33 receptor expressed on the surface of CHO cells and Daudi cells. The results in Table 2 and Figures 3 A-3F further demonstrate, that the human TSPAN33 antibodies 11E8, S24, and S3 do not bind other TSPAN family members TSPAN5, TSPAN14 or TSPAN17. Table 2. Cell surface binding of human TSPAN33 antibodies

Binding to activated B cells: In vitro binding activity of the exemplified human TSPAN33 antibodies to human TSPAN33 on activated PBMCs from healthy human volunteers is evaluated by flow cytometry. PBMCs are isolated by standard density centrifugation (Ficoll-Paque) methods, resuspended at 6 x 10⁷ in 45 mL media, and stimulated overnight with recombinant CD40L (20 ng/mL) and recombinant IL-4 (20 ng/mL) in a 37 °C, 5% CO2 incubator. The next day, PBMCs are washed, incubated with Fc blocker at 1:500 (Thermo Fisher 14-9161-73) for 10 min, and plated at 3 x 10⁵ PBMCs/well in a 96-well round bottom plate. Plates are then spun down, media is gently removed, and cells are stained with human TSPAN33 antibodies (fluorescently labeled with Alexa Fluor 647 Thermo Fisher A20186) in 1:500 Fc blocker, 50ul of DPBS + 2% FCS for 30min at 4C. As controls, activated cells are stained with hlgGl and PE anti-human TSPAN33 (BAAM) antibody (Biolegend 395403). Stained cells are washed and analyzed on a flow cytometer. The binding of human TSPAN33 antibodies is represented as the percentage of antibody that bound the CD20⁺ B cells human and percentage of cells that bound CD4⁺ T cells in the flow cytometry analysis.

The results as demonstrated in Figure 5 show that the Fc and CDR modified human

TSPAN33 antibodies S3-C2-XEQ and S24-C2-IEQ specifically bind to activated CD20⁺ B ceils and not to CD4⁺ T cells.

Example 3. Effector Function Activity of the human TSPAN33 antibodies

Antibody dependent cellular cytotoxicity (ADCC) In vitro ADCC assays of the exemplified human TSPAN33 antibodies are conducted using Daudi cells (ATCC #CCL-213) expressing human TSPAN33 and human CD20, and with Jurkat cells expressing functional FcyRIIIa (V158)-NFAT-Luc (Eli Lilly and Company) as the effector cell line. Daudi target cells either activated (activated overnight with IL-4 and anti-CD40L) or unactivated (resting) are plated at 40,000 celis/welL with the exemplified human TSPAN33 antibodies, and CD20 positive control antibody (such as described in U.S. Pat. No. 8,153,125), ail of which are serially diluted down 2~fold from 3.3 pg/mL to 26 ng/mL, and incubated for 1 hour at 37 °C. Next, Jurkat VI 58 cells are added to the wells at 200,000 cells/well and incubated for 3 hours at 37 °C, followed by addition of 150 pL/well of Bright Glo Luciferase substrate (Promega, p/n. E2610). The contents of the plates are mixed using a plate shaker at low speed, for 10 minutes and the luminescence signal is read on a Spec traM ax L microplate reader (Molecular Devices). The data are exported into Sigmaplot and the relative luminescence units (RLU) for each antibody concentration are plotted in a scatter format of antibody concentration versus RLU.

The results in Figures 6A-6C and Figures 7A-7B show that the exemplified human TSPAN33 antibodies can induce ADCC activity in both unactivated (resting) and activated Daudi cells expressing TSPAN33. Specifically, Figures 6A-6C (unactivated Daudi cells) and 7A-7B (activated Daudi cells) show that the human TSPAN33 parental antibodies 11E8, S3, S24 and Fc and CDR modified antibodies 11E8-Y92H-IEQ and S24-C2-IEQ induce an ADCC response in both the unactivated and activated Daudi cells when compared to the control IgG. Furthermore, the results as demonstrated in Figures 6B-6C and 7A-7B, show that the Fc and CDR modified human TSPAN33 antibodies 11E8-Y92H- IEQ and S24-C2-IEQ induce an enhanced ADCC response when compared to the parental 11E8 and S24 antibodies respectively with both the unactivated and activated Daudi cells. Complement dependent cellular cytotoxicity ( CDC) In vitro CDC assays of the exemplified human TSPAN33 antibodies are conducted using human or mouse full-length TSPAN33 transfected CHO cells, or human Daudi cells. Cells to be assayed are collected, washed and resuspended to a final concentration of 1 ^c 10⁶ cells/mL in CDC assay buffer (RPMI-1640 with no phenol red (Coming 17-105-CV) supplemented with 0.1% BSA (Sigma A7906)), and plated at 5 ^c 10⁴ cells at 50 pL/well in a 96 well clear bottom black tissue culture treated plate (BD Falcon # 353219), Plates are then incubated at 37 °C 5% CCh in a cell culture incubator. The exemplified human TSPAN33 antibodies and CD20 antibody (positive control) are serially diluted from 30 pg/mL in CDC assay buffer and added at 50 pL/well to the plated cells in the appropriate wells and incubated at 37 °C for 30 minutes. Controls are included in duplicate: cells alone, cells plus complement, and cells plus CD20 antibody. Next, 100 pL of CDC assay buffer is added to the cells alone control and 50 pL is added to the other controls. Human serum complement (Quidel, #A133) quickly thawed in a 37 °C water bath is diluted 1 :5 in CDC assay media and added at 50 pL/well to the assay plate. The plate is gently agitated and incubated for 3 hours at 37 °C 5% CO2 cell culture incubator. Next, 20 pL/well of Cell Titer Aqueous Solution (MTS) (Promega G3582) is added and the plate is incubated overnight at 37 °C. The following day, fluorescence (RFU) at 490 nM is read using a Molecular Devices Spectra Max Gemini EM plate reader.

The results as demonstrated in Figure 8, show that the human TSPAN33 antibodies may induce CDC in the human TSPAN33 transfected CHO cell line in a concentration- dependent manner.

SEQUENCE LISTING

4D4

SEQ ID NO: 1 4D4 HCDR1 (North)

KASGYTFTSHFIH

SEQ ID NO: 2 4D4 HCDR2 (North)

IIRPGGGSSRHAQKFLD

SEQ ID NO: 3 4D4 HCDR3 (North)

AREVIGTEYPFDY

SEQ ID NO: 4 4D4 LCDR1 (North)

RASQSISNWLA

SEQ ID NO: 5 4D4 LCDR2 (North)

YKASSLES

SEQ ID NO: 6 4D4 LCDR3 (North)

QQYNSFYT

SEQ ID NO: 7 4D4 HC Variable Region (VH)

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSHFIHWVRQAPGQGLEWMGIIRPG GGS SRH AQKFLDR VTMTRDT S T S T VYMEL S SLRSEDT A V Y Y CARE VIGTE YPFD Y WGQGTLVTVSS

SEQ ID NO: 8 4D4 LC Variable Region (VL)

DIQMTQSPSTLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYKASSLES

GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSFYTFGQGTKLEIK

SEQ ID NO: 9 4D4 HC

QVQLVQSGAEVKKPGASVKVSCKASGYTFTSHFIHWVRQAPGQGLEWMGIIRPG GGS SRH AQKFLDR VTMTRDT S T S T VYMEL S SLRSEDT A V Y Y C ARE VIGTE YPFD Y W GQGTL VT V S S AS TKGP S VFPL AP S SK S T S GGT A ALGCL VKD YFPEP VT V S WN S G ALTSGVHTFP AVLQ S SGL Y SL S S VVTVP S S SLGTQT YICNVNHKP SNTKVDKRVEP KSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCKV SN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK

SEQ ID NO: 10 4D4 LC

DIQMTQSPSTLSASVGDRVTITCRASQSISNWLAWYQQKPGKAPKLLIYKASSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSFYTFGQGTKLEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YSL S S TLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 11 4D4 HC DNA

CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG

TGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCCACTTCATACAC

T GGGT GC GAC AGGCCC CTGGAC A AGGGC TT GAGT GG AT GGG A AT A AT C AGGC

CTGGTGGTGGTAGTTCAAGGCACGCACAGAAGTTCCTGGACAGAGTCACCAT

GACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGA

T C T G AGG AC AC GGC C GT GT AT T AC T GT GC G AG AG A AGT A ATT GGG AC C G A AT

ACCCTTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCC

ACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGG

GGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTG

ACGGTGTCGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGG

CTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCC

TCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCA

GCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCA

CACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCC

TCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTC

ACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACT

GGT ATGT GGACGGCGTGGAGGT GC AT AAT GCC AAGAC AAAGCCGCGGGAGG AGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCA

AG AC T GGC T G A AT GGC A AGG AGT AC A AGT GC A AGGT C TC C A AC A A AGC C C TC

CCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAAC

CACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGT

CAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGT

GGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCT

GGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCA

GGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCA

CAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA

SEQ ID NO: 12 4D4 LC DNA

GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAG

AGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAACTGGTTGGCCTGGT

ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCGTCTAG

TTT AGAAAGT GGGGTCCC ATC AAGGTT C AGC GGC AGT GGATCTGGGAC AGAA

TTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTG

CCAACAATATAATAGTTTTTACACTTTTGGCCAGGGGACCAAGCTGGAGATCA

AAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAG

TTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAG

AGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCC

CAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGC

AGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCT

GCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAG

GGGAGAGT GC

S3

SEQ ID NO: 13 S3 HCDR1 (North)

KASGYTFTSYFMH

SEQ ID NO: 14 S3 HCDR2 (North)

IIKL Y GGSTRNAQKF QG SEQ ID NO: 15 S3 HCDR3 (North)

ARESNW GRAFDI

SEQ ID NO: 16 S3 / S3-C2-IEQ LCDR1 (North)

RASQSISTWLA

SEQ ID NO: 17 S3 LCDR2 (North)

YKASSLES

SEQ ID NO: 18 S3 LCDR3 (North)

QQYNSFYT

SEQ ID NO: 19 S3 VH

EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYFMHWVRQAPGQGLEWMGIIKL

YGGSTRNAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARESNWGRAF

DIWGQGTMVTVSS

SEQ ID NO: 20 S3 VL

DIVMTQSPSTLSASVGDRVTITCRASQSISTWLAWSQQKPGKAPKLLIYKASSLES

GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSFYTFGQGTKVEIK

SEQ ID NO: 21 S3 HC

EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYFMHWVRQAPGQGLEWMGIIKL YGGSTRNAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARESNWGRAF DIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFP AVLQ S SGL Y SL S S VVTVP S S SLGTQT YICNVNHKP SNTKVDKR VEPKSCDKTHT CPPCP APELLGGP S VFLFPPKPKDTLMISRTPEVT C V VVD V SHED PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPP VLD SDGSFFL Y SKLT VDKSRWQQGNVF SC S VMHE AL HNHYTQKSLSLSPGK SEQ ID NO: 22 S3 LC

DIVMTQSPSTLSASVGDRVTITCRASQSISTWLAWSQQKPGKAPKLLIYKASSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSFYTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YSL S S TLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 23 S3 HC DNA

G AGGT GC AGC T GGT GC AGT C T GGGGC T G AGGT G A AG A AGC C T GGGGC C T C AG

TGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTTTATGCAC

T GGGT GC GAC AGGCCC CTGGAC A AGGGC TT GAGT GG AT GGG A AT A AT C A A AC

TTTATGGTGGTAGCACAAGGAACGCACAGAAGTTCCAGGGCAGAGTCACCAT

GACC AGGGAC ACGTCC ACGAGC AC AGTCT AC AT GGAGTT GAGC AGCC TGAGA

TCTGAGGACACGGCCGTGTATTACTGTGCGAGAGAGTCTAACTGGGGACGGG

CTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCCTCAGCCTCCACC

AAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGG

CACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACG

GTGTCGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTG

TCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCC

AGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCA

ACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACAC

ATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCT

TCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCAC

AT GCGT GGT GGT GGACGT GAGC C AC GA AGAC CC T GAGGT C A AGTT C A ACTGG

TATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAG

CAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAG

ACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCC

AGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCA

CAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCA

GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGG

GAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGG

ACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGG TGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACA

ACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA

SEQ ID NO: 24 S3 LC DNA

GACATCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTTGGAGACAG

AGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTACCTGGTTGGCCTGGT

CTCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATAAGGCGTCTAG

TTT AGAAAGT GGGGTCCC AT C AAGGTTC AGCGGC AGTGGATCTGGGAC AGA A

TTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTG

C C A AC AGT AT A AT AGTTTTT AC AC TTTT GGC C AGGGGAC C A AGGT GGA A AT C

AAAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCA

GTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCA

GAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTC

CCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAG

CAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCC

TGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACA

GGGGAGAGTGC

S3-C2-IEO

SEQ ID NO: 25 S3-C2-IEQ HCDR1 (North)

KASGGTFTSYFMH

SEQ ID NO: 26 S3-C2-IEQ HCDR2 (North)

IIKL Y GGPTRNAQKF QG

SEQ ID NO: 27 S3-C2-IEQ HCDR3 (North)

ARESNWDRAFDI

SEQ ID NO: 16 S3 / S3-C2-IEQ LCDR1 (North)

RASQSISTWLA SEQ ID NO: 28 S3-C2-IEQ LCDR2 (North)

YKVSSLES

SEQ ID NO: 29 S3-C2-IEQ LCDR3 (North)

GQYNSFYA

SEQ ID NO: 30 S3-C2-IEQ VH

QVQLVQSGAEVKKPGASVKVSCKASGGTFTSYFMHWVRQAPGQGLEWMGIIKL

Y GGPTRNAQKF QGRVTMTRDTSTSTVYMELS SLRSEDT AVYY C ARESNWDRAF DIWGQGTMVTVSS

SEQ ID NO: 31 S3-C2-IEQ VL

DIQMTQSPSTLSASVGDRVTITCRASQSISTWLAWYQQKPGKAPKLLIYKVSSLES GVP SRF S GS GS GTEFTLTIS SLQPDDF AT Y Y C GQ YN SF Y AF GQGTK VEIK

SEQ ID NO: 32 S3-C2-IEQ HC

QVQLVQSGAEVKKPGASVKVSCKASGGTFTSYFMHWVRQAPGQGLEWMGIIKL

Y GGPTRNAQKF QGRVTMTRDTSTSTVYMELS SLRSEDT AVYY C ARESNWDRAF DIWGQGTM VT V S S ASTKGP S VFPL AP S SKST SGGT AALGCL VKD YFPEP VT V S WN SGALTSGVHTFP AVLQ S SGL Y SL S S VVTVP S S SLGTQT YICNVNHKP SNTKVDKK VEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKIKDTLMISRTPEVTCVVVDVSHED PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCK VSNKALPAPEEKTISKQKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPP VLD SDGSFFL Y SKLT VDKSRWQQGNVF SC S VMHE AL HNHYTQKSLSLSPG

SEQ ID NO: 33 S3-C2-IEQ LC

DIQMT Q SP STL S AS VGDRVTITCRASQ SISTWL AW Y QQKPGK APKLLIYK V S SLES GVPSRF SGSGSGTEFTLTIS SLQPDDF AT YYCGQYN SF YAFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KD S T YSL S S TLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC SEQ ID NO: 34 S3-C2-IEQ HC DNA

C AGGT AC A ATT GGT GC AGT C AGGAGC AGAAGT GAAAAAGCCTGGGGC A AGC

GTGAAAGTGAGTTGCAAAGCTAGCGGCGGTACTTTTACATCTTATTTCATGCA

T T GGGT G AG AC AGGC T C C T GGT C AGGG AC T GG A AT GG AT GGGC AT A AT C A AG

TTGTATGGCGGCCCCACTCGGAATGCACAGAAATTCCAAGGGAGGGTGACTA

TGACTAGAGATACAAGTACTTCCACCGTTTATATGGAATTGAGTAGCCTTCGT

AGCGAGGACACCGCCGTGTACTATTGCGCCCGTGAAAGCAACTGGGATCGTG

CATTTGACATCTGGGGACAAGGCACAATGGTCACCGTTTCTAGTGCCTCCACC

AAGGGCCCATCGGTCTTCCCGCTAGCACCCTCCTCCAAGAGCACCTCTGGGGG

CACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACG

GTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGT

CCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCA

GCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAA

CACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACA

TGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTT

CCCCCCAAAAATCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACA

T GC GT GGT GGT GG AC GT G AGC C AC G A AG AC C C T G AGGT C A AGT T C A AC T GGT

AC GT GG AC GGC GT GG AGGT GC AT A AT GC C A AG AC A A AGC C GC GGG AGGAGC

AGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGA

CTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA

GCCCCCGAGGAGAAAACCATCTCCAAACAGAAAGGGCAGCCCCGAGAACCA

CAGGTGTACACCCTGCCCCCATCCCGGGACGAGCTGACCAAGAACCAGGTCA

GCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGG

GAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCCCCCGTGCTG

GACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAG

GTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

SEQ ID NO: 35 S3-C2-IEQ LC DNA

GATATACAAATGACCCAGTCACCAAGCACACTCTCCGCATCCGTAGGGGATC GTGTAACAATCACATGCAGAGCTTCCCAGTCTATCTCTACCTGGCTCGCATGG T ATC A AC AAAAACC AGGT AAGGC ACC AAAGTT GTT GAT AT AC AAGGTTTCTT CTCTGGAGTCCGGGGTTCCAAGTCGTTTTAGCGGCAGCGGATCAGGCACCGA

GTTCACTCTGACCATATCAAGCCTCCAGCCAGATGATTTTGCCACTTATTATT

GCGGACAATACAACAGTTTCTATGCTTTTGGGCAGGGTACTAAAGTCGAGAT

CAAGCGAACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGC

AGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCC

AGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACT

CCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA

GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC

CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC

AGGGGAGAGT GC

S24

SEQ ID NO: 36 S24 / S24-C2-IEQ / S24-IEQ HCDR1 (North)

QGSGYRFSSYWIG

SEQ ID NO: 37 S24 / S24-C2-IEQ / S24-IEQ HCDR2 (North)

IIYPGDSDTRYSPSFQG

SEQ ID NO: 38 S24 / S24-IEQ HCDR3 (North)

ARHNYYY GMD V

SEQ ID NO: 39 S24 / S24-C2-IEQ / S24-IEQ LCDR1 (North)

RASQSLSSSFLA

SEQ ID NO: 40 S24 / S24-C2-IEQ / S24-IEQ LCDR2 (North)

YDASRRAT

SEQ ID NO: 41 S24 / S24-IEQ LCDR3 (North)

HQYGRSYT SEQ ID NO: 42 S24 VH

EVQLVQSGAEVKKPGESLKISCQGSGYRFSSYWIGWVRQMPGKGLEWMGIIYPG

DSDTRYSPSFQGQVTFSADKSISKVYLQWSSLKASDTAIYFCARHNYYYGMDVW

GQGTTVTVSS

SEQ ID NO: 43 S24 VL

EIVLTQSPGTLSLSPGERATLSCRASQSLSSSFLAWYQQRPDQAPRLLIYDASRRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A VYHCHQ Y GRS YTF GQ GTK VEIK

SEQ ID NO: 44 S24 HC

EVQLVQSGAEVKKPGESLKISCQGSGYRFSSYWIGWVRQMPGKGLEWMGIIYPG DSDTRYSPSFQGQVTFSADKSISKVYLQWSSLKASDTAIYFCARHNYYYGMDVW GQGTT VT V S S A STKGP S VFPL AP S SK S T S GGT A ALGCL VKD YFPEP VT V S WN S GA LTSGVHTFP AVLQ S SGL Y SL S S VVTVP S S SLGTQT YICNVNHKP SNTKVDKRVEPK SCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNW YVDGVEVHNAKTKPREEQ YNST YRVV S VLTVLHQDWLN GKEYKCK V SNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK

SEQ ID NO: 45 S24 LC

EIVLTQSPGTLSLSPGERATLSCRASQSLSSSFLAWYQQRPDQAPRLLIYDASRRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A VYHCHQ Y GRS YTF GQ GTK VEIKRT V A A PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKD STYSLSSTLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 46 S24 HC DNA

GAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTC TGAAGATCTCCTGTCAGGGTTCTGGATATAGGTTTTCCAGTTACTGGATCGGC TGGGTGCGCCAGATGCCCGGGAAAGGCCTGGAGTGGATGGGGATCATCTATC CTGGTGACTCTGATACCAGATACAGTCCGTCCTTCCAAGGCCAGGTCACCTTC T C AGC CGAC A AGT C A AT C AGT A A AGTCT ATTT GC AGT GGAGT AGCC T GA AGG CCTCGGACACCGCCATATATTTCTGTGCGAGACATAACTACTACTACGGCATG

GACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCTCCACCAAGG

GCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACA

GCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT

CGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCT

ACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCA

GCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACAC

CAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGC

CCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCC

CCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGC

GTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTATG

TGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGT

ACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAAGACTG

GCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG

GTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAAGTCAGCC

TGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA

GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGAC

TCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGAGCAGGTG

GCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAAC

CACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA

SEQ ID NO: 47 S24 LC DNA

GAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAG

AGCCACCCTCTCCTGCAGGGCCAGTCAGAGTCTTAGCAGCAGCTTCTTAGCCT

GGTACCAGCAGAGACCTGACCAGGCTCCCAGACTCCTCATCTATGATGCATCC

AGAAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGGTCTGGGACAG

ACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATCAC

T GTC ATC AGT AT GGTCGTT CAT AC ACTTTT GGCC AGGGGACC AAGGT GGAGAT

CAAAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGATGAGC

AGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCC

AGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACT CCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGT GC

S24-C2-IEO

SEQ ID NO: 36 S24 / S24-C2-IEQ / S24-IEQ HCDR1 (North)

QGSGYRFSSYWIG

SEQ ID NO: 37 S24 / S24-C2-IEQ / S24-IEQ HCDR2 (North)

IIYPGDSDTRYSPSFQG

SEQ ID NO: 48 S24-C2-IEQ HCDR3 (North)

YRHNYYYAQDV

SEQ ID NO: 39 S24 / S24-C2-IEQ / S24-IEQ LCDR1 (North)

RASQSLSSSFLA

SEQ ID NO: 40 S24 / S24-C2-IEQ / S24-IEQ LCDR2 (North)

YDASRRAT

SEQ ID NO: 49 S24-C2-IEQ LCDR3 (North)

HQYPRSYT

SEQ ID NO: 50 S24-C2-IEQ VH

EVQLVQSGAEVKKPGESLKISCQGSGYRFSSYWIGWVRQMPGKGLEWMGIIYPG

DSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCYRHNYYYAQDV

WGQGTTVTVSS

SEQ ID NO: 51 S24-C2-IEQ VL

EIVLTQSPGTLSLSPGERATLSCRASQSLSSSFLAWYQQKPGQAPRLLIYDASRRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQ YPRS YTF GQGTK VEIK SEQ ID NO: 52 S24-C2-IEQ HC

EVQLVQSGAEVKKPGESLKISCQGSGYRFSSYWIGWVRQMPGKGLEWMGIIYPG

DSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCYRHNYYYAQDV

WGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFP AVLQ S SGL Y SL S S VVTVP S S SLGTQT YICNVNHKP SNTKVDKK VE

PKSCDKTHTCPPCPAPELLGGPSVFLFPPKIKDTLMISRTPEVTCVVVDVSHEDPE

VKFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCKV S

NKALPAPEEKTISKQKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE

WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH

NHYTQKSLSLSPG

SEQ ID NO: 53 S24-C2-IEQ LC

EIVLTQSPGTLSLSPGERATLSCRASQSLSSSFLAWYQQKPGQAPRLLIYDASRRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQ YPRS YTF GQGTK VEIKRT V A A PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKD STYSLSSTLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 54 S24-C2-IEQ HC DNA

GAGGT AC AACTTGTTC AATC AGGAGCTGAAGTT AAA AAACCTGGCGAAAGTT

TGAAGATCTCATGTCAGGGCTCAGGATATAGATTTAGCAGCTACTGGATCGGT

T GGGT AC GCC A AAT GCCC GGC A AGGGGC T GGA AT GGAT GGGGATT AT AT AT C

CTGGCGACAGTGATACTCGGTACTCCCCTAGTTTTCAGGGCCAAGTCACAATT

TCAGCCGATAAGTCAATAAGCACAGCCTACCTCCAATGGAGCAGTCTTAAAG

CATCAGACACTGCCATGTATTATTGTTATAGACACAATTATTACTACGCCCAG

GATGTATGGGGCCAGGGCACAACAGTAACTGTTTCTTCCGCCTCCACCAAGG

GCCCATCGGTCTTCCCGCTAGCACCCTCCTCCAAGAGCACCTCTGGGGGCACA

GCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT

CGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTA

CAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAG

CTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACC

AAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCC CACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCC

CCAAAAATCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCG

T GGT GGT GG AC GT G AGC C AC G A AG AC C C T G AGGT C A AGT T C A AC T GGT AC GT

GGAC GGCGT GGAGGT GC AT A AT GC C A AG AC A A AGC CGC GGGAGGAGC AGT A

CAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGG

CTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC

CCGAGGAGAAAACCATCTCCAAACAGAAAGGGCAGCCCCGAGAACCACAGG

TGTACACCCTGCCCCCATCCCGGGACGAGCTGACCAAGAACCAGGTCAGCCT

GACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAG

AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCCCCCGTGCTGGACT

CCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACC

ACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

SEQ ID NO: 55 S24-C2-IEQ LC DNA

GAGATCGTGTTGACTCAGTCACCTGGTACACTGTCCTTGTCACCAGGGGAGAG

GGCTACCCTGTCATGCAGGGCTTCACAGTCTTTGAGTAGTTCATTCTTGGCTT

GGTATCAGCAAAAACCCGGCCAAGCACCAAGGCTGCTTATATACGATGCCTC

CCGGCGGGCCACTGGTATCCCTGACCGATTTTCTGGGTCCGGTTCAGGAACTG

ACTTCACACTCACCATAAGCCGCTTGGAACCAGAGGACTTCGCAGTCTATTAC

TGTCACCAATACCCTAGATCATACACATTCGGCCAGGGAACCAAGGTGGAGA

TAAAACGAACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAG

CAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCC

C AGAGAGGCC AAAGT AC AGT GGAAGGT GGAT AACGCCCTCC AATCGGGT AAC

TCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA

GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC

CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC

AGGGGAGAGT GC

11E8

SEQ ID NO: 56 11E8 / 11E8-Y92H-IEQ HCDR1 (North)

KSSGYTFTSYYIH SEQ ID NO: 57 11E8 / 11E8-Y92H-IEQ HCDR2 (North)

IINPRGGISISAQKFRG SEQ ID NO: 58 11E8 / 11E8-Y92H-IEQ HCDR3 (North)

ARGDN SGWYPLFD S

SEQ ID NO: 59 11E8 / 11E8-Y92H-IEQ LCDR1 (North)

RASQSVSSSYLA

SEQ ID NO: 60 11E8 / 11E8-Y92H-IEQ LCDR2 (North)

FGASSRAT

SEQ ID NO: 61 11E8 LCDR3 (North) HQYVNSPLT

SEQ ID NO: 62 11E8 / 11E8-Y92H-IEQ VH

QVQLVQSGAEVKKPGASVKVSCKSSGYTFTSYYIHWVRQAPGQGLEWMGIINPR GGISISAQKFRGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDNSGWYPLFD SWGQGTLVTVSS

SEQ ID NO: 63 11E8 VL

EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIFGASSRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQ YVN SPLTF GGGTK VEIK

SEQ ID NO: 64 11E8 HC

QVQLVQSGAEVKKPGASVKVSCKSSGYTFTSYYIHWVRQAPGQGLEWMGIINPR GGISISAQKFRGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDNSGWYPLFD S W GQGTL VT V S S A S TKGP S VFPL AP S SK S T SGGT A ALGCL VKD YFPEP VT V S WN S GALT S GVHTFP A VLQ S S GL Y SL S SWT VP S S SLGTQT YICNVNHKP SNTK VDKRV EPKSCDKTHT CPPCP APELLGGP S VFLFPPKPKDTLMISRTPEVT C VVVD V SHEDP EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCKV SNKALP APIEKTISK AKGQPREPQ VYTLPP SREEMTKN Q V SLTCL VKGF YP SDIAV

EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH

NHYTQKSLSLSPGK

SEQ ID NO: 65 11E8 LC

EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIFGASSRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQ YVN SPLTF GGGTK VEIKRT V A APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ D SKD S T YSL S S TLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 66 11E8 HC DNA

CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG

TGAAGGTTTCCTGCAAGTCGTCTGGATACACCTTCACCAGCTACTATATACAC

T GGGT GC GAC AGGCCC CTGGAC A AGGGC TT GAGT GG AT GGG A AT A AT C A ACC

CTAGGGGTGGTATTTCAATCTCCGCACAGAAGTTCCGGGGCAGAGTCACCAT

GACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGA

TCTGAGGACACGGCCGTGTATTACTGTGCGAGAGGGGATAACAGTGGCTGGT

ACCCCCTCTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCC

TCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC

TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG

GTGACGGTGTCGTGGAACTCAGGCGCACTGACCAGCGGCGTGCACACCTTCC

CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG

CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC

CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC

TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT

TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG

GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA

ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA

GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC

CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC

TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA

ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA

GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT

GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA

GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT

GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA

SEQ ID NO: 67 11E8 LC DNA

GAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAG

AGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCT

GGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTTTGGTGCATCC

AGC AGGGCC ACTGGC ATCCC AGAC AGGTT C AGT GGC AGT GGGTCTGGGAC AG

ACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTAC

TGTCACCAGTATGTTAACTCACCGCTCACTTTCGGCGGAGGGACCAAGGTGG

AGATCAAAAGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTA

TCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGT

AACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGC

CTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCT

ACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTT

C AAC AGGGGAGAGT GC

11E8-Y92H-IEO

SEQ ID NO: 56 11E8 / 11E8-Y92H-IEQ HCDR1 (North)

KSSGYTFTSYYIH

SEQ ID NO: 57 11E8 / 11E8-Y92H-IEQ HCDR2 (North)

IINPRGGISISAQKFRG

SEQ ID NO: 58 11E8 / 11E8-Y92H-IEQ HCDR3 (North)

ARGDN SGW YPLFD S SEQ ID NO: 59 11E8 / 11E8-Y92H-IEQ LCDR1 (North)

RASQSVSSSYLA

SEQ ID NO: 60 11E8 / 11E8-Y92H-IEQ LCDR2 (North)

FGASSRAT

SEQ ID NO: 68 11E8-Y92H-IEQ LCDR3 (North)

HQHVNSPLT

SEQ ID NO: 62 11E8 / 11E8-Y92H-IEQ VH

QVQLVQSGAEVKKPGASVKVSCKSSGYTFTSYYIHWVRQAPGQGLEWMGIINPR

GGISISAQKFRGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDNSGWYPLFD

SWGQGTLVTVSS

SEQ ID NO: 69 11E8-Y92H-IEQ VL

EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIFGASSRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQHVN SPLTF GGGTK VEIK

SEQ ID NO: 70 11E8-Y92H-IEQ HC

QVQLVQSGAEVKKPGASVKVSCKSSGYTFTSYYIHWVRQAPGQGLEWMGIINPR GGISISAQKFRGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGDNSGWYPLFD S W GQGTL VT V S S A STKGP S VFPL AP S SK S T S GGT A ALGCL VKD YFPEP VT V S WN S GALT S GVHTFP A VLQ S S GL Y SL S SWT VP S S SLGTQT YICNVNHKP SNTK VDKK V EPKSCDKTHT CPPCP APELLGGP S VFLFPPKIKDTLMISRTPEVT C VVVD V SHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCKV S NKALPAPEEKTISKQKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG

SEQ ID NO: 71 11E8-Y92H-IEQ LC

EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIFGASSRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQHVN SPLTF GGGTK VEIKRTV A APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ D SKD S T YSL S S TLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 72 11E8-Y92H-IEQ HC DNA

CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG

TGAAGGTTTCCTGCaAGTCGTCTGGATACACCTTCACCAGCTACTATATACACT

GGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAATAATCAACCC

TAGGGGTGGTATTTCAATCTCCGCACAGAAGTTCCGGGGCAGAGTCACCATG

ACCAGGGACACGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGAT

CTGAGGACACGGCCGTGTATTACTGTGCGAGAGGGGATAACAGTGGCTGGTA

CCCCCTCTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCT

CCACCAAGGGCCCATCGGTCTTCCCGCTAGCACCCTCCTCCAAGAGCACCTCT

GGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGG

TGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCC

GGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGC

CCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCC

CAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACT

CACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTT

CCTCTTCCCCCCAAAAATCAAGGACACCCTCATGATCTCCCGGACCCCTGAGG

T C AC AT GC GT GGT GGT GG AC GT G AGC C AC G A AG AC C C T G AGGT C A AGTT C A A

CTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA

GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC

CAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC

TCCCAGCCCCCGAGGAGAAAACCATCTCCAAACAGAAAGGGCAGCCCCGAG

AACCACAGGTGTACACCCTGCCCCCATCCCGGGACGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGG

AGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCCCCCG

TGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAG

AGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTC

TGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT SEQ ID NO: 73 11E8-Y92H-IEQ LC DNA

GAAATTGTGTTGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAG

AGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCT

GGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTTTGGTGCATCC

AGC AGGGCC ACTGGC ATCCC AGAC AGGTT C AGT GGC AGT GGGTCTGGGAC AG

ACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTAC

TGTCACCAGCATGTTAACTCACCGCTCACTTTCGGCGGAGGGACCAAGGTGG

AGATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGAT

GAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTA

TCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGT

AACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGC

CTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCT

ACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTT

C AAC AGGGGAGAGT GC

S24-IEO

SEQ ID NO: 36 S24 / S24-C2-IEQ / S24-IEQ HCDR1 (North)

QGSGYRFSSYWIG

SEQ ID NO: 37 S24 / S24-C2-IEQ / S24-IEQ HCDR2 (North)

IIYPGDSDTRYSPSFQG

SEQ ID NO: 38 S24 / S24-IEQ HCDR3 (North)

ARHNYYY GMD V

SEQ ID NO: 39 S24 / S24-C2-IEQ / S24-IEQ LCDR1 (North)

RASQSLSSSFLA

SEQ ID NO: 40 S24 / S24-C2-IEQ / S24-IEQ LCDR2 (North)

YDASRRAT SEQ ID NO: 41 S24 / S24-IEQ LCDR3 (North)

HQYGRSYT

SEQ ID NO: 74 S24-IEQ VH

EVQLVQSGAEVKKPGESLKISCQGSGYRFSSYWIGWVRQMPGKGLEWMGIIYPG

DSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARHNYYYGMDV

WGQGTTVTVSS

SEQ ID NO: 75 S24-IEQ VL

EIVLTQSPGTLSLSPGERATLSCRASQSLSSSFLAWYQQKPGQAPRLLIYDASRRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQ Y GRS YTF GQ GTK VEIK

SEQ ID NO: 76 S24-IEQ HC

EVQLVQSGAEVKKPGESLKISCQGSGYRFSSYWIGWVRQMPGKGLEWMGIIYPG

DSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARHNYYYGMDV

WGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG

ALTSGVHTFP AVLQ S SGL Y SL S S VVTVP S S SLGTQT YICNVNHKP SNTKVDKK VE

PKSCDKTHTCPPCPAPELLGGPSVFLFPPKIKDTLMISRTPEVTCVVVDVSHEDPE

VKFNWYVDGVEVHNAKTKPREEQYNSTYRVV S VLTVLHQDWLNGKEYKCKV S

NKALPAPEEKTISKQKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE

WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH

NHYTQKSLSLSPG

SEQ ID NO: 77 S24-IEQ LC

EIVLTQSPGTLSLSPGERATLSCRASQSLSSSFLAWYQQKPGQAPRLLIYDASRRA T GIPDRF S GS GS GTDF TLTISRLEPEDF A V Y Y CHQ Y GRS YTF GQ GTK VEIKRT V A A PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKD STYSLSSTLTL SK AD YEKHK V Y ACE VTHQGL S SP VTK SFNRGEC

SEQ ID NO: 78 S24-IEQ HC DNA

GAGGTACAACTTGTTCAATCAGGAGCTGAAGTTAAAAAACCTGGCGAAAGTT

TGAAGATCTCATGTCAGGGCTCAGGATATAGATTTAGCAGCTACTGGATCGGT T GGGT AC GCC A AAT GCCC GGC A AGGGGC T GGA AT GGAT GGGGATT AT AT AT C

CTGGCGACAGTGATACTCGGTACTCCCCTAGTTTTCAGGGCCAAGTCACAATT

TCAGCCGATAAGTCAATAAGCACAGCCTACCTCCAATGGAGCAGTCTTAAAG

CATCAGACACTGCCATGTATTATTGTGCTAGACACAATTATTACTACGGCATG

GATGTATGGGGCCAGGGCACAACAGTAACTGTTTCTTCCGCCTCCACCAAGG

GCCCATCGGTCTTCCCGCTAGCACCCTCCTCCAAGAGCACCTCTGGGGGCACA

GCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGT

CGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTA

CAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAG

CTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACC

AAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCC

CACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCC

CCAAAAATCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCG

T GGT GGT GGACGT GAGC C AC GA AGACC CTGAGGT C A AGTT C A ACTGGT AC GT

GGAC GGCGT GGAGGT GC AT AAT GC C A AG AC A A AGC CGC GGGAGGAGC AGT A

CAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGG

CTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCC

CCGAGGAGAAAACCATCTCCAAACAGAAAGGGCAGCCCCGAGAACCACAGG

TGTACACCCTGCCCCCATCCCGGGACGAGCTGACCAAGAACCAGGTCAGCCT

GACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAG

AGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCCCCCGTGCTGGACT

CCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACC

ACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

SEQ ID NO: 79 S24-IEQ LC DNA

GAGATCGTGTTGACTCAGTCACCTGGTACACTGTCCTTGTCACCAGGGGAGAG

GGCTACCCTGTCATGCAGGGCTTCACAGTCTTTGAGTAGTTCATTCTTGGCTT

GGTATCAGCAAAAACCCGGCCAAGCACCAAGGCTGCTTATATACGATGCCTC

CCGGCGGGCCACTGGTATCCCTGACCGATTTTCTGGGTCCGGTTCAGGAACTG

ACTTCACACTCACCATAAGCCGCTTGGAACCAGAGGACTTCGCAGTCTATTAC

T GTC ACC A AT AC GGT AG AT CAT AC AC ATTCGGCC AGGGAACC A AGGTGGAGA TAAAACGAACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAG CAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCC C AGAGAGGCC AAAGT AC AGT GGAAGGT GGAT AACGCCCTCC AATCGGGT AAC TCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCA GCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGC CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGT GC

SEQ ID NO: 80 Human TSPAN33 M ARRPRAP AASGEEF SF V SPLVKYLLFFFNMLF W VISMVM VAV GV Y ARLMKHA AALACLAVDPAILLIVVGVLMFLLTFCGCIGSLRENICLLQTFSLCLTAVFLLQLA AGILGFVF SDKARGK V SEIINNAIVHYRDDLDLQNLIDFGQKKF SCCGGIS YKDW S QNMYFNC SEDNPSRERC S VP Y SCCLPTPDQ AVINTMCGQGMQ AFDYLEASKVIY TNGCIDKLVNWIHSNLFLLGGVALGLAIPQLVGILLSQILVNQIKDQIKLQLYNQQ HRADPWY

Claims

1. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO:l, the HCDR2 comprises SEQ ID NO:2, the HCDR3 comprises SEQ ID NO:3, the LCDR1 comprises SEQ ID NO:4, the LCDR2 comprises SEQ ID NO:5, and the LCDR3 comprises SEQ ID NO:6.

2. The antibody of claim 1, wherein the VH comprises SEQ ID NO: 7 and the VL comprises SEQ ID NO: 8.

3. The antibody of claim 1 or 2, wherein the antibody comprises a heavy chain (HC) comprising SEQ ID NO: 9 and a light chain (LC) comprising SEQ ID NO: 10.

4. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 13, the HCDR2 comprises SEQ ID NO: 14, the HCDR3 comprises SEQ ID NO: 15, the LCDR1 comprises SEQ ID NO: 16, the LCDR2 comprises SEQ ID NO: 17, and the LCDR3 comprises SEQ ID NO: 18.

5. The antibody of claim 4, wherein the VH comprises SEQ ID NO: 19 and the VL comprises SEQ ID NO: 20.

6. The antibody of claim 4 or 5, wherein the antibody comprises a HC comprising SEQ ID NO: 21 and a LC comprising SEQ ID NO: 22.

7. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 25, the HCDR2 comprises SEQ ID NO: 26, the HCDR3 comprises SEQ ID NO: 27, the LCDR1 comprises SEQ ID NO: 16, the LCDR2 comprises SEQ ID NO: 28, and the LCDR3 comprises SEQ ID NO: 29.

8. The antibody of claim 7, wherein the VH comprises SEQ ID NO: 30 and the VL comprises SEQ ID NO: 31.

9. The antibody of claim 7 or 8, wherein the antibody comprises a HC comprising SEQ ID NO: 32 and a LC comprising SEQ ID NO: 33.

10. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 36, the HCDR2 comprises SEQ ID NO: 37, the HCDR3 comprises SEQ ID NO: 38, the LCDR1 comprises SEQ ID NO: 39, the LCDR2 comprises SEQ ID NO: 40, and the LCDR3 comprises SEQ ID NO: 41.

11. The antibody of claim 10, wherein the VH comprises SEQ ID NO: 42 or 74, and the VL comprises SEQ ID NO: 43 or 75.

12. The antibody of claim 10, wherein the antibody comprises a HC comprising SEQ ID NO: 44 or 76, and a LC comprising SEQ ID NO: 45 or 77.

13. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 36, the HCDR2 comprises SEQ ID NO: 37, the HCDR3 comprises SEQ ID NO: 48, the LCDR1 comprises SEQ ID NO: 39, the LCDR2 comprises SEQ ID NO: 40, and the LCDR3 comprises SEQ ID NO: 49.

14. The antibody of claim 13, wherein the VH comprises SEQ ID NO: 50 and the VL comprises SEQ ID NO: 51.

15. The antibody of claim 13 or 14, wherein the antibody comprises a HC comprising SEQ ID NO: 52 and a LC comprising SEQ ID NO: 53.

16. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 56, the HCDR2 comprises SEQ ID NO: 57, the HCDR3 comprises SEQ ID NO: 58, the LCDR1 comprises SEQ ID NO: 59, the LCDR2 comprises SEQ ID NO: 60, and the LCDR3 comprises SEQ ID NO: 61.

17. The antibody of claim 16, wherein the VH comprises SEQ ID NO: 62 and the VL comprises SEQ ID NO: 63.

18. The antibody of claim 16 or 17, wherein the antibody comprises a HC comprising SEQ ID NO: 64 and a LC comprising SEQ ID NO: 65.

19. An antibody that specifically binds human TSPAN33, wherein the antibody comprises a VH and a VL, wherein the VH comprises HCDR1, HCDR2, and HCDR3, and the VL comprises LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 56, the HCDR2 comprises SEQ ID NO: 57, the HCDR3 comprises SEQ ID NO: 58, the LCDR1 comprises SEQ ID NO: 59, the LCDR2 comprises SEQ ID NO: 60, and the LCDR3 comprises SEQ ID NO: 68.

20. The antibody of claim 19, wherein the VH comprises SEQ ID NO: 62 and the VL comprises SEQ ID NO: 69.

21. The antibody of claim 19 or 20, wherein the antibody comprises a HC comprising SEQ ID NO: 70 and a LC comprising SEQ ID NO: 71.

22. The antibody of any one of claims 1 to 21, wherein the antibody binds activated B cells, and wherein the antibody is a depleting antibody.

23. A nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 21, 22, 32, 33, 44, 45, 52, 53, 64, 65, 70, 71, 76, or 77.

24. A vector comprising the nucleic acid of claim 23.

25. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 9, and a second nucleic acid sequence encoding SEQ ID NO: 10

26. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 21, and a second nucleic acid sequence encoding SEQ ID NO: 22.

27. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 32, and a second nucleic acid sequence encoding SEQ ID NO: 33.

28. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 44, and a second nucleic acid sequence encoding SEQ ID NO: 45.

29. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 52, and a second nucleic acid sequence encoding SEQ ID NO: 53.

30. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 64, and a second nucleic acid sequence encoding SEQ ID NO: 65.

31. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 70, and a second nucleic acid sequence encoding SEQ ID NO: 71.

32. The vector of claim 24, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 76, and a second nucleic acid sequence encoding SEQ ID NO: 77.

33. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10.

34. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 21, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 22.

35. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 32, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 33.

36. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 44, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 45.

37. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 52, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 53.

38. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 64, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 65.

39. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 70, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 71.

40. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 76, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 77.

41. A cell comprising the vector of any one of claims 33 to 40.

42. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10.

43. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 21, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 22.

44. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 32, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 33.

45. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 44, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 45.

46. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 52, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 53.

47. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 64, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 65.

48. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 70, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 71.

49. A cell comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 76, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 77.

50. The cell of any one of claims 41 to 49, wherein the cell is a mammalian cell.

51. A process of producing an antibody comprising culturing the cell of any one of claims 41 to 50, under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.

52. An antibody produced by the process of claim 51.

53. A pharmaceutical composition comprising the antibody of any one of Claims 1 to 22 or 52, and a pharmaceutically acceptable excipient, diluent, or carrier.

54. A method of treating a B cell associated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody of any one of Claims 1 to 22 or 52, or the pharmaceutical composition of claim 53.

55. The method of Claim 54, wherein the B cell associated disorder comprises activated B cells.

56. The method of Claim 54, wherein the B cell associated disorder is an autoimmune disease.

57. The method of Claim 54, wherein the B cell associated disorder is selected from Systemic Lupus Erythematosus, Lupus Nephritis, Multiple Sclerosis, Rheumatoid Arthritis, Granulomatosis with Polyangiitis, Wegener’s Granulomatosis, Microscopic Polyangiitis, Pemphigus Vulgaris, Type 1 diabetes, Sjogren's Syndrome, or Myasthenia Gravis.

58. The method of Claim 54, wherein the B cell associated disorder is cancer.

59. The method of Claim 58, wherein the cancer is B cell lymphoma.

60. The method of Claim 58, wherein the cancer is Non-Hodgkin’s Lymphoma, or Chronic Lymphocytic Leukemia.

61. The method of Claim 58, further comprising administering one or more chemotherapeutic agents to the subject.

62. The method of Claim 61, wherein the chemotherapeutic agents are administered in simultaneous, separate, or sequential combination with the antibody or pharmaceutical composition thereof.

63. The method of claim 61 or 62, wherein the chemotherapeutic agent comprises fludarabine or cyclophosphamide.

64. The method of Claim 58, further comprising administering ionizing radiation in simultaneous, separate, or sequential combination with the antibody or pharmaceutical composition thereof.

65. The antibody of any one of Claims 1 to 22 or 52 for use in therapy.

66. The antibody of any one of Claims 1 to 22 or 52, or the pharmaceutical composition of claim 49, for use in the treatment of a B cell associated disorder.

67. The antibody or pharmaceutical composition for use of Claim 66, wherein the B cell associated disorder comprises activated B cells.

68. The antibody or pharmaceutical composition for use of Claim 66, wherein the B cell associated disorder is an autoimmune disease.

69. The antibody or pharmaceutical composition for use of claim 66, wherein the B cell associated disorder is selected from Systemic Lupus Erythematosus, Lupus Nephritis, Multiple Sclerosis, Rheumatoid Arthritis, Granulomatosis with Polyangiitis, Wegener’s Granulomatosis, Microscopic Polyangiitis, Pemphigus Vulgaris, Type 1 diabetes, Sjogren's Syndrome, or Myasthenia Gravis.

70. The antibody or pharmaceutical composition for use of claim 66, wherein the B cell associated disorder is cancer.

71. The antibody or pharmaceutical composition for use of Claim 70, wherein the cancer is B cell lymphoma.

72. The antibody or pharmaceutical composition for use of Claim 70, wherein the cancer is Non-Hodgkin’s Lymphoma, or Chronic Lymphocytic Leukemia.

73. The antibody or pharmaceutical composition for use of Claim 70 further comprising one or more chemotherapeutic agents to be used in combination with the antibody or pharmaceutical composition thereof.

74. The antibody or pharmaceutical composition for use of Claim 73, wherein the chemotherapeutic agents are for simultaneous, separate, or sequential administration with the antibody or pharmaceutical composition thereof.

75. The antibody or pharmaceutical composition for use of Claim 73 or 74, wherein the chemotherapeutic agent comprises fludarabine or cyclophosphamide.

76. The antibody or pharmaceutical composition for use of Claim 70, further comprising ionizing radiation to be used in simultaneous, separate, or sequential combination with the antibody or pharmaceutical composition thereof.

77. Use of the antibody of any one of claims 1 to 22 or 52, in the manufacture of a medicament for the treatment of a B cell associated disorder.

78. The use of claim 77, wherein the B cell associated disorder is an autoimmune disease.

79. The use of claim 77, wherein the B cell associated disorder comprises activated B cells.

80. The use of claim 77, wherein the B cell associated disorder is selected from systemic lupus erythematosus, multiple sclerosis, rheumatoid arthritis, Sjogren’s syndrome, idiopathic thrombocytopenia purpura, Type 1 diabetes, pemphigus vulgaris, neuromyelitis optica, ANCA vasculitis, or myasthenia gravis.

81. The use of claim 77, wherein the B cell associated disorder is cancer.

82. The use of claim 81, wherein the cancer is B cell lymphoma.

83. The use of claim 81, wherein the cancer is Non-Hodgkin’s Lymphoma, or Chronic Lymphocytic Leukemia.