Antibodies binding to human PAD4 and uses thereof

Abstract

The present application relates to a specific anti-PAD4 (PAD4), a specific anti-PAD4 inhibitor, a specific anti-PAD4 inhibitor, and a specific anti-PAD4 inhibitor. The present invention relates to peptidyl arginine deiminase 4) antibodies, nucleic acids encoding the antibodies, vectors and host cells comprising the nucleic acids, and methods of making and using the antibodies.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 369,184, filed on July 22, 2022, and U.S. Provisional Application No. 63 / 397,698, filed on August 12, 2022, both of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present application relates to specific anti-PAD4 (peptidyl arginine deiminase 4) antibodies, nucleic acids encoding the antibodies, vectors and host cells comprising the nucleic acids, and methods of making and using the antibodies. Background Art

[0004] PAD4 (peptidyl arginine deiminase 4) is one of five PAD enzymes (PAD1, 2, 3, 4, and 6) in humans and is present in several immune cell types and works by converting arginine to citrulline. (Chavanas et al., Gene 330:19-27 (2004)). This citrullination is a stress response and can act as a signal to remove stressed cells. (Brentville et al., Oncoimmunology 8:e1576490 (2019)). In neutrophils, PAD4 also plays a role in a process called NETosis, through which neutrophils extrude a decondensed chromatin structure complex containing DNA scaffolds, citrullinated histones, and antimicrobial neutrophil granules. (Li et al. J. Exp. Med. 207:1853-62 (2010)). These extruded complexes are called neutrophil extracellular traps (NETs), and during NETosis, these NETs capture and kill invading microorganisms as part of the innate immune response. (Chamardani et al., Mol. Cell. Biochem. 477: 673-88 (2022)). A similar process involving monocytes is called METosis and involves the formation of monocyte extracellular traps (METs). In addition, proteins citrullinated by PAD4 become antigenic substrates and are targets of cellular (i.e., T cell) and humoral (i.e., B cell-derived antibodies) adaptive immune responses. (See, e.g., Curran et al. Nat. Rev. Rheumatol. 16: 301-15 (2020); Brentville et al.) Therefore, PAD4 activity can lead to the production of anti-citrullinated protein antibodies (ACPAs).

[0005] Various data indicate that PAD4 plays a role in diseases such as rheumatoid arthritis (RA), lupus (including systemic lupus erythematosus (SLE), lupus nephritis, vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, inflammatory bowel disease (IBD) (including ulcerative colitis and Crohn's disease), thrombosis (such as venous thrombosis), antiphospholipid antibody syndrome, cystic fibrosis, and cancer. (See, e.g., Curran et al.; Yadav et al., J. Cyst. Fibros. 18:636-45 (2018); Wang et al., Front. Immunol. 13:895-216 (2022); Fresneda ... Alarcon et al. Frong. Immunol. 12:649693 (2021); Weeding et al. Clin. Immunol. 196:110-116 (2018); Xu et al. Chinese J. Microbiology and Immunology 12:115-121 (2020); Yoshida et al. Clin. Kidney J. 6:308-12 (2013); O'Sullivan et al. Rheumatology, 58 (Suppl. 2):kez061.024 (2019); Pan et al. Authora Preprints, 2021, DOI: 10.22541 / au.161590650.07168461 / v1; D. Zhu et al., Pharmaceutics, 8: 14(11): 2414(2022). For example, rheumatoid arthritis (RA) is a major autoimmune disorder involving tissue (i.e., joint and non-joint) and systemic inflammation. The progression of RA is associated with elevated levels of autoantibodies, and approximately 70% of RA patients are positive for anti-citrullinated protein antibodies (ACPA). ACPAs appear, for example, in the lungs and blood of people at risk for RA, and high ACPA levels are associated with severe RA symptoms (such as bone erosion) and certain complications (such as cardiovascular disease). In addition, the presence of ACPAs worsens RA symptoms in a murine model of spontaneous arthritis.

[0006] ACPA recognizes neoantigens, particularly neoantigens from proteins citrullinated by PAD enzymes. PAD4 is the main PAD enzyme found in synovial tissue. For example, PAD4 can mediate ACPA-dependent and ACPA-independent functions that are important for the pathogenesis of RA and other diseases. In addition, approximately 15% of RA patients have antibodies that activate PAD4. The presence of such PAD4-activating antibodies is associated with severe joint erosion disease. At the genetic level, single nucleotide polymorphisms (SNPs) in the PADI4 gene encoding PAD4 have been identified as susceptibility haplotypes that promote RA. (Suzuki et al., Nat. Genet. 34:395-402 (2003)). In addition, epigenetic changes in the PADI4 promoter region are associated with lower disease activity in RA patients compared to healthy individuals. (Kolarz et al., Clin. Med. 9(7):2049 (2020)). RA patients carrying the PADI4 susceptibility haplotype were found to have higher titers of anti-citrullinated peptide antibodies (ACPA) than RA patients who did not carry this haplotype (Reyes-Castillo et al., Clin. Exp. Immunol. 182(2):119-31 (2015)).

[0007] Therefore, blocking the activity of PAD4 can be used to treat RA and to treat individuals at risk for developing RA as well as other inflammatory and autoimmune diseases or other diseases associated with NETosis, METosis, the presence of anti-citrullinated protein antibodies (ACPA), increased PAD4 expression, or increased PAD4 activity (such as increased polypeptide citrullination).

[0008] The present invention provides anti-PAD4 antibodies that inhibit PAD4 activity, nucleic acids encoding the antibodies, vectors and host cells containing the antibodies, and methods for producing and using the antibodies. Summary of the Invention

[0009] The present invention relates to specific anti-PAD4 antibodies, nucleic acids encoding the antibodies, vectors and host cells comprising the nucleic acids, and methods of making and using the antibodies. For example, embodiments of the present invention include the following:

[0010] 1. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4 or 62 and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7.

[0011] 2. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4 or 62, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 9.

[0012] 3. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 221 or 225 and the amino acid sequence of SEQ ID NO: 222; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 223.

[0013] 4. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 221 or 225 and the amino acid sequence of SEQ ID NO: 222; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 223, and further wherein the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 224 or 235.

[0014] 5. The isolated antibody of any one of embodiments 1 to 4, wherein the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NO: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68; and wherein the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NO: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70.

[0015] 6. The isolated antibody of any one of embodiments 1 to 4, wherein the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68 modified by 1 to 10 amino acid substitutions; and wherein the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0016] 7. The isolated antibody of any one of embodiments 1 to 6, wherein the heavy chain variable region comprises a glycine at Kabat position 94, which position corresponds to position 98 of SEQ ID NO: 10.

[0017] 8. The isolated antibody of any one of embodiments 1 to 7, wherein the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NO: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68.

[0018] 9. The isolated antibody of any one of embodiments 1 to 8, wherein the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NO: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70.

[0019] 10. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 10; and a VL comprising the amino acid sequence of SEQ ID NO: 12.

[0020] 11. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 14; and a VL comprising the amino acid sequence of SEQ ID NO: 16.

[0021] 12. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 18; and a VL comprising the amino acid sequence of SEQ ID NO: 20.

[0022] 13. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 22; and a VL comprising the amino acid sequence of SEQ ID NO: 24.

[0023] 14. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 26; and a VL comprising the amino acid sequence of SEQ ID NO: 28.

[0024] 15. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 30; and a VL comprising the amino acid sequence of SEQ ID NO: 32.

[0025] 16. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 34; and a VL comprising the amino acid sequence of SEQ ID NO: 36.

[0026] 17. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 38; and a VL comprising the amino acid sequence of SEQ ID NO: 40.

[0027] 18. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 42; and a VL comprising the amino acid sequence of SEQ ID NO: 44.

[0028] 19. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 46; and a VL comprising the amino acid sequence of SEQ ID NO: 48.

[0029] 20. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 50; and a VL comprising the amino acid sequence of SEQ ID NO: 52.

[0030] 21. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 54; and a VL comprising the amino acid sequence of SEQ ID NO: 56.

[0031] 22. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 58; and a VL comprising the amino acid sequence of SEQ ID NO: 60.

[0032] 23. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 68; and a VL comprising the amino acid sequence of SEQ ID NO: 70.

[0033] 24. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 233; and a VL comprising the amino acid sequence of SEQ ID NO: 234.

[0034] 25. The isolated antibody of any one of embodiments 1 to 23, wherein the antibody binds to an epitope on PAD4 comprising SEQ ID NO: 217 and SEQ ID NO: 218.

[0035] 26. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 73, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 74; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 75, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 76, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 77.

[0036] 27. The isolated antibody of embodiment 26, wherein the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NO: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134; and wherein the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136.

[0037] 28. The isolated antibody of embodiment 26, wherein the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134, modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and wherein the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NOs: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136, modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0038] 29. The isolated antibody of embodiment 25, 26 or 27, wherein the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NO: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130 or 134.

[0039] 30. The isolated antibody of any one of embodiments 26 to 29, wherein the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136.

[0040] 31. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 78; and a VL comprising the amino acid sequence of SEQ ID NO: 80.

[0041] 32. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 82; and a VL comprising the amino acid sequence of SEQ ID NO: 84.

[0042] 33. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 86; and a VL comprising the amino acid sequence of SEQ ID NO: 88.

[0043] 34. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 90; and a VL comprising the amino acid sequence of SEQ ID NO: 92.

[0044] 35. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 94; and a VL comprising the amino acid sequence of SEQ ID NO: 96.

[0045] 36. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 98; and a VL comprising the amino acid sequence of SEQ ID NO: 100.

[0046] 37. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 102; and a VL comprising the amino acid sequence of SEQ ID NO: 104.

[0047] 38. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 106; and a VL comprising the amino acid sequence of SEQ ID NO: 108.

[0048] 39. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 110; and a VL comprising the amino acid sequence of SEQ ID NO: 112.

[0049] 40. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 114; and a VL comprising the amino acid sequence of SEQ ID NO: 116.

[0050] 41. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 118; and a VL comprising the amino acid sequence of SEQ ID NO: 120.

[0051] 42. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 122; and a VL comprising the amino acid sequence of SEQ ID NO: 124.

[0052] 43. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 126; and a VL comprising the amino acid sequence of SEQ ID NO: 128.

[0053] 44. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 130; and a VL comprising the amino acid sequence of SEQ ID NO: 132.

[0054] 45. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 134; and a VL comprising the amino acid sequence of SEQ ID NO: 136.

[0055] 46. ​​The isolated antibody of any one of embodiments 26 to 45, wherein the antibody binds to an epitope on PAD4 comprising SEQ ID NO: 219 and SEQ ID NO: 220.

[0056] 47. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1), the HCDR1 comprising the following amino acid sequence:

[0057] positions 26 to 35 of SEQ ID NO: 138; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 138; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 138;

[0058] positions 26 to 35 of SEQ ID NO: 140; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 140; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 140;

[0059] positions 26 to 35 of SEQ ID NO: 142; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 142; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 142;

[0060] positions 26 to 35 of SEQ ID NO: 144; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 144; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 144;

[0061] positions 26 to 35 of SEQ ID NO: 146; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 146; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 146;

[0062] positions 26 to 35 of SEQ ID NO: 148; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 148; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 148;

[0063] positions 26 to 35 of SEQ ID NO: 150; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 150; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 150;

[0064] positions 26 to 35 of SEQ ID NO: 152; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 152; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 152;

[0065] positions 26 to 35 of SEQ ID NO: 154; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 154; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 154;

[0066] positions 26 to 35 of SEQ ID NO: 156; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 156; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 156;

[0067] positions 26 to 35 of SEQ ID NO: 158; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 158; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 158;

[0068] positions 26 to 35 of SEQ ID NO: 160; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 160; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 160;

[0069] positions 26 to 35 of SEQ ID NO: 162; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 162; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 162;

[0070] positions 26 to 35 of SEQ ID NO: 164; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 164; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 164;

[0071] positions 26 to 35 of SEQ ID NO: 166; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 166; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 166;

[0072] positions 26 to 35 of SEQ ID NO: 168; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 168; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 168; or

[0073] positions 26 to 35 of SEQ ID NO: 170; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 170; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 170; and

[0074] The antibody comprises a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170.

[0075] 48. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1), the HCDR1 comprising the following amino acid sequence:

[0076] positions 26 to 35 of SEQ ID NO: 138; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 138; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 138;

[0077] positions 26 to 35 of SEQ ID NO: 140; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 140; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 140;

[0078] positions 26 to 35 of SEQ ID NO: 142; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 142; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 142;

[0079] positions 26 to 35 of SEQ ID NO: 144; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 144; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 144;

[0080] positions 26 to 35 of SEQ ID NO: 146; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 146; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 146;

[0081] positions 26 to 35 of SEQ ID NO: 148; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 148; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 148;

[0082] positions 26 to 35 of SEQ ID NO: 150; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 150; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 150;

[0083] positions 26 to 35 of SEQ ID NO: 152; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 152; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 152;

[0084] positions 26 to 35 of SEQ ID NO: 154; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 154; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 154;

[0085] positions 26 to 35 of SEQ ID NO: 156; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 156; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 156;

[0086] positions 26 to 35 of SEQ ID NO: 158; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 158; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 158;

[0087] positions 26 to 35 of SEQ ID NO: 160; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 160; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 160;

[0088] positions 26 to 35 of SEQ ID NO: 162; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 162; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 162;

[0089] positions 26 to 35 of SEQ ID NO: 164; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 164; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 164;

[0090] positions 26 to 35 of SEQ ID NO: 166; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 166; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 166; or

[0091] positions 26 to 35 of SEQ ID NO: 168; a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 168; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 168; and

[0092] The antibody comprises a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172.

[0093] 49. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of residues 26 to 35 of SEQ ID NO: 168, a HCDR2 comprising the amino acid sequence of residues 50 to 66 of SEQ ID NO: 168, and a HCDR comprising the amino acid sequence of residues 99 to 108 of SEQ ID NO: 168; and wherein the antibody comprises a light chain variable region (VL) comprising: a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172.

[0094] 50. The isolated antibody of embodiment 49, wherein the antibody binds to PAD4 at pH 6.0 with lower affinity compared to an antibody comprising a VH comprising the amino acid sequence of SEQ ID NO: 30 and a VL comprising the amino acid sequence of SEQ ID NO: 32.

[0095] 51. The isolated antibody of any one of embodiments 1 to 50, which is an IgA, IgG or IgM antibody.

[0096] 52. The isolated antibody of any one of embodiments 1 to 50, which is an IgG antibody, such as a human IgG1, IgG2, IgG3, or IgG4 antibody or a murine IgG1 or IgG2 antibody.

[0097] 53. The isolated antibody of embodiment 52, wherein the antibody comprises a wild-type, human IgG1, IgG2, or IgG4 heavy chain constant region.

[0098] 54. The isolated antibody of any one of embodiments 1 to 50, wherein the antibody comprises a human IgG1 or IgG2 heavy chain constant region comprising an amino acid sequence that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and / or wherein the antibody comprises a light chain constant region comprising an amino acid sequence that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194.

[0099] 55. The isolated antibody of any one of embodiments 1 to 50, wherein the antibody comprises a human IgG1 or IgG2 heavy chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192, and / or comprises a light chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194.

[0100] 56. The isolated antibody of any one of embodiments 1 to 55, wherein the antibody comprises a human IgG1 or IgG2 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192.

[0101] 57. The isolated antibody of any one of embodiments 1 to 56, wherein the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.

[0102] 58. The isolated antibody of any one of embodiments 1 to 57, which is a full-length antibody.

[0103] 59. The isolated antibody of any one of embodiments 1 to 57, which is an IgG antibody lacking a C-terminal lysine in the heavy chain constant region.

[0104] 60. The isolated antibody of any one of embodiments 1 to 50, which is an antibody fragment, such as Fv, single-chain Fv (scFv), Fab, Fab' or (Fab')2.

[0105] 61. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 68 followed by the amino acid sequence of SEQ ID NO: 178; and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 70 followed by the amino acid sequence of SEQ ID NO: 194.

[0106] 62. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 68, followed by the amino acid sequence of SEQ ID NO: 180; and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 70, followed by the amino acid sequence of SEQ ID NO: 194.

[0107] 63. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a HC comprising the amino acid sequence of SEQ ID NO: 196 and a light chain comprising the amino acid sequence of SEQ ID NO: 200.

[0108] 64. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a HC comprising the amino acid sequence of SEQ ID NO: 198 and a light chain comprising the amino acid sequence of SEQ ID NO: 200.

[0109] 65. The isolated antibody of any one of embodiments 1 to 64, which is a bispecific or multispecific antibody, or which is covalently or non-covalently bound to at least one other molecule.

[0110] 66. The isolated antibody of embodiment 65, wherein the antibody is covalently or non-covalently bound to at least one other molecule, wherein the at least one other molecule comprises a detectable label and / or a drug.

[0111] 67. An isolated antibody that specifically binds to murine protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of positions 31 to 35 of SEQ ID NO: 208, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 208, and a HCDR3 comprising the amino acid sequence of positions 99 to 107 of SEQ ID NO: 208; and wherein the antibody comprises a light chain variable region (VL) comprising: a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 210, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 210, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 210.

[0112] 68. The isolated antibody of embodiment 67, wherein the antibody comprises a human IgG1 heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO: 208; and / or wherein the antibody comprises a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO: 210.

[0113] 69. The isolated antibody of embodiment 67, wherein the antibody comprises a VH amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 208; and / or wherein the antibody comprises a VL amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 210.

[0114] 70. The isolated antibody of any one of embodiments 67 to 69, wherein the antibody comprises the VH amino acid sequence of SEQ ID NO: 208 and / or the VL amino acid sequence of SEQ ID NO: 210.

[0115] 71. The isolated antibody of any one of embodiments 67 to 70, wherein the antibody comprises: a heavy chain comprising an amino acid sequence that is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO: 210, and / or wherein the antibody comprises: a light chain comprising an amino acid sequence that is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO: 212.

[0116] 72. The isolated antibody of any one of embodiments 67 to 71, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 212, and / or wherein the antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 214.

[0117] 73. The isolated antibody of any one of embodiments 1 to 66, wherein the antibody does not bind to human protein arginine deiminase 2 (PAD2).

[0118] 74. The isolated antibody of any one of embodiments 1 to 66 or 73, wherein the antibody inhibits the conversion of arginine to citrulline in the peptide substrate TSTGGRQGSHH (SEQ ID NO: 216) by PAD4 with an IC50 of 10 to 200 nM, 50 to 200 nM, 10 to 100 nM, 20 to 100 nM or 50 to 100 nM.

[0119] 75. The isolated antibody of any one of embodiments 1 to 66, 73 or 74, wherein the antibody optionally inhibits the conversion of a peptide substrate to PAD4 in a dose-dependent manner with an IC50 of 0.1 to 10 nM (such as 0.2 to 5 nM). Arginine was converted to citrulline in the culture medium with PAD4 concentrations ranging from 1 to 8 μg / mL.

[0120] 76. The isolated antibody of any one of embodiments 1 to 66 or 73 to 75, wherein the antibody inhibits the deamination of Na-benzoyl-L-arginine ethyl ester hydrochloride (BAEE) by PAD4.

[0121] 77. The isolated antibody of any one of embodiments 1 to 66 or 73 to 76, wherein the antibody has a K of less than 5 nM, less than 3 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.05 nM to 1 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.1 nM, or 0.5 nM to 1 nM. D Binds specifically to PAD4.

[0122] 78. The isolated antibody of embodiment 77, wherein the antibody has a K of less than 5 nM, less than 3 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.05 nM to 1 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.1 nM, or 0.5 nM to 1 nM. DSpecifically binds to PAD4 (a) at 37°C in the presence of 1 to 2 mM calcium chloride (e.g., 1 mM calcium chloride) and / or (b) at 37°C in the absence of additional Ca 2+ and measured by surface plasmon resonance (SPR) in the presence of 2 mM EDTA.

[0123] 79. The isolated antibody of any one of embodiments 1 to 66 or 73 to 78, wherein the antibody has an ECM score of less than 50, less than 30, less than 10, less than 5, 1 to 30, 1 to 20, 1 to 10, 1 to 5, 1 to 3, 1, 2, 3, 4, or 5 in an ECM assay.

[0124] 80. The isolated antibody of any one of embodiments 1 to 66 or 73 to 79, wherein the immunogenicity of the antibody is less than one, two, or all three of the following as measured by an in silico immunogenicity assay: (alemtuzumab), (rituximab), (daclizumab).

[0125] 81. The isolated antibody of any one of embodiments 1 to 66 or 73 to 80, wherein the antibody reduces the amount of extracellular citrullinated histone H3 and / or reduces the secretion of GM-CSF and / or gene expression of GM-CSF in LPS-stimulated human blood monocytes compared to an isotype control antibody.

[0126] 82. The isolated antibody of any one of embodiments 1 to 66 or 73 to 81, wherein the antibody is capable of being internalized by LPS-stimulated human blood monocytes.

[0127] 83. The isolated antibody of any one of embodiments 1 to 82, wherein the antibody inhibits PAD4 function in inflamed lungs, e.g., as demonstrated by reduced citrullination (e.g., H3) in bronchoalveolar lavage fluid (BALF) collected from the lungs.

[0128] 84. The isolated antibody of any one of embodiments 1 to 83, wherein the antibody inhibits the function of PAD4 in inflamed joints, eg, as evidenced by reduced citrullination of ITIH4 and / or PRG4 in joint tissue.

[0129] 85. The isolated antibody of any one of embodiments 1 to 84, wherein the antibody reduces citrullination (eg, H3) in neutrophils, monocytes, M1 macrophages, and / or M2 macrophages compared to an isotype control antibody.

[0130] 86. The isolated antibody of any one of embodiments 1 to 66 or 73 to 85, wherein the antibody reduces the amount of extracellular citrullination (e.g., H3 and / or ITIH4 citrullination) in the BALF in an LPS-induced acute lung inflammation model in human PAD4 knock-in mice, optionally wherein the amount of citrullinated H3 is reduced by at least 15%, at least 30%, at least 40%, at least 50% or at least 60%, and / or wherein the amount of citrullinated ITIH4 is reduced by at least 50%, at least 70% or at least 80%, compared to an isotype control antibody.

[0131] 87. The isolated antibody of any one of embodiments 1 to 66 or 73 to 86, wherein the antibody reduces the amount of extracellular citrullination (e.g., PRG4 and / or ITIH4 citrullination) in mouse patella in an LPS-induced acute joint injury model in human PAD4 knock-in mice compared to an isotype control antibody, optionally wherein the EC50 for reducing citrullinated ITIH4 and / or citrullinated PRG4 is 2 nM or less, 1 nM or less, 0.1 nM or less, 0.05 to 2 nM, or 0.1 to 1 nM.

[0132] 88. A pharmaceutical composition comprising the antibody of any one of embodiments 1 to 87 and a pharmaceutically acceptable carrier.

[0133] 89. An isolated nucleic acid or a collection of two or more nucleic acids encoding the antibody of any one of embodiments 1 to 87.

[0134] 90. An isolated vector comprising one or more nucleic acids encoding the heavy and light chains of the antibody of any one of embodiments 1 to 87.

[0135] 91. An isolated host cell comprising the nucleic acid of embodiment 89 or the vector of embodiment 90.

[0136] 92. A method for producing an antibody that specifically binds to PAD4, comprising culturing the host cell of embodiment 91 under conditions suitable for expression of the antibody.

[0137] 93. The method of embodiment 92, further comprising recovering the antibody from the host cell.

[0138] 94. An antibody produced by the method of embodiment 92 or 93.

[0139] 95. Use of the antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88 for the preparation of a medicament for the treatment of an autoimmune disorder.

[0140] 96. The use according to embodiment 95, wherein the autoimmune disorder is rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or another autoimmune disorder described herein.

[0141] 97. The isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88, for use in treating an autoimmune disorder.

[0142] 98. The use as used in embodiment 97, wherein the autoimmune disorder is rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or another autoimmune disorder described herein.

[0143] 99. A method of treating an autoimmune disorder in a subject in need thereof, comprising administering to the subject an effective amount of the isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88.

[0144] 100. The method of embodiment 99, wherein the autoimmune disorder is rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or another autoimmune disorder described herein.

[0145] 101. The use, the antibody for use, or the method according to any one of embodiments 95 to 100, wherein the method or use comprises administering at least one additional therapeutic agent, optionally wherein the at least one additional therapeutic agent is one or more of methotrexate, adalimumab, etanercept, infliximab, hydroxychloroquine, sulfasalazine, leflunomide, abatacept, anakinra, certolizumab, golimumab, rituximab, sarilumab, tocilizumab, baricitinib, tofacitinib, or upadacitinib.

[0146] Other exemplary embodiments include antibodies as described in any one of embodiments 1 to 87, which comprise at least one post-translational modification of the amino acid sequence. In some embodiments, the modification is modification of the N-terminal Glu or Gln to pyroglutamate. In some such embodiments, the light chain N-terminal Glu or Gln is modified to pyroglutamate.

[0147] The disclosure herein also encompasses the use of an antibody as described in any one of embodiments 1 to 87 above or a pharmaceutical composition as described in embodiment 88 above for the preparation of a medicament for treating an individual at risk of developing RA; and the isolated antibody as described in any one of embodiments 1 to 87 above or a pharmaceutical composition as described in embodiment 88 above for treating an individual at risk of developing RA. The present invention also encompasses a method of treating an individual at risk of developing RA, comprising administering to the individual an effective amount of an isolated antibody as described in any one of embodiments 1 to 87 above or a pharmaceutical composition as described in embodiment 88. In some cases, the individual has one or more of the following conditions: (a) at least one first-degree relative (e.g., a parent or sibling) with RA; (b) the presence of anti-citrullinated protein antibodies (ACPA) in the serum; (c) the presence of rheumatoid factor (RF) in the serum; (d) arthralgia in at least one joint; (e) inflammation in at least one joint observed by ultrasound or magnetic resonance imaging (MRI); or (f) undifferentiated arthritis. In some cases, the individual is in remission. In some cases, the method or use comprises administering at least one additional therapeutic agent, optionally wherein the at least one additional therapeutic agent is one or more of: methotrexate, adalimumab, etanercept, infliximab, hydroxychloroquine, sulfasalazine, leflunomide, abatacept, anakinra, certolizumab pegol, golimumab, rituximab, salimumab, tocilizumab, baricitinib, tofacitinib, or upapeptinib.

[0148] In other embodiments, the antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88 can be used to prepare a medicament for inhibiting NETosis or METosis in a subject. In some embodiments, the subject is a subject with an autoimmune disorder such as rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or a subject at risk of developing an autoimmune disease (e.g., rheumatoid arthritis). Other embodiments include the isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition according to claim 88 for inhibiting NETosis or METosis in a subject. In some embodiments, the subject is an individual with an autoimmune disorder such as rheumatoid arthritis, lupus, lupus nephritis, vasculitis or thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or another autoimmune disorder disclosed herein. In some embodiments, the subject is an individual at risk for developing an autoimmune disorder (e.g., rheumatoid arthritis). Other embodiments also include a method of inhibiting NETosis or METosis in an individual, the method comprising administering to the individual an effective amount of an antibody as described in any one of embodiments 1 to 87 or a pharmaceutical composition as described in embodiment 88. In some embodiments, the subject is an individual with an autoimmune disorder such as rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or another autoimmune disorder disclosed herein. In some embodiments, the individual is an individual at risk of developing an autoimmune disorder (e.g., rheumatoid arthritis). In some cases, the method or use comprises administering at least one other therapeutic agent, optionally wherein the at least one other therapeutic agent is one or more of: methotrexate, adalimumab, etanercept, infliximab, hydroxychloroquine, sulfasalazine, leflunomide, abatacept, anakinra, certolizumab pegol, golimumab, rituximab, salimumab, tocilizumab, baricitinib, tofacitinib, or upapatinib. The disclosure herein also encompasses a method for inhibiting NETosis or METosis in a biological sample in vitro, comprising administering to the biological sample an effective amount of an isolated antibody according to any one of embodiments 1 to 87 or a pharmaceutical composition according to embodiment 88.The methods and uses herein also include a method of inhibiting citrullination in a subject, comprising administering to the subject an effective amount of an isolated antibody according to any one of embodiments 1 to 87, or use of an isolated antibody according to any one of embodiments 1 to 87 for inhibiting citrullination in a subject, or use of an isolated antibody according to any one of embodiments 1 to 87 for the preparation of a medicament for inhibiting citrullination in a subject. The present invention also includes a method of inhibiting citrullination in a biological sample in vitro, comprising administering to the biological sample an effective amount of an isolated antibody according to any one of embodiments 1 to 87.

[0149] In other embodiments, the antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88 can be used to prepare a medicament for preventing the onset or recurrence of an autoimmune disorder such as rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (ulcerative colitis, Crohn's disease), or another autoimmune disorder disclosed herein, or the isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition according to claim 88 can be used to prevent the onset or recurrence of an autoimmune disorder such as rheumatoid arthritis, lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease), or another autoimmune disorder disclosed herein. Yet other embodiments include a method of preventing the onset or recurrence of an autoimmune disorder (such as rheumatoid arthritis, lupus, lupus nephritis, vasculitis, thrombosis (e.g., venous thrombosis), inflammatory bowel disease (IBD), colitis, ulcerative colitis), or another autoimmune disorder disclosed herein in a subject in need thereof, comprising administering to the subject an effective amount of an isolated antibody according to any one of embodiments 1 to 87 or a pharmaceutical composition according to embodiment 88. In some embodiments, the subject has been determined to be susceptible to the onset or recurrence of the disorder. In some such uses and methods herein, the antibody is administered to the subject together with another therapeutic agent.

[0150] In another embodiment, the isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88 can be used to prepare a medicament for treating cancer, or the isolated antibody or pharmaceutical composition can be used to treat cancer. Other embodiments also include a method of treating cancer in an individual in need thereof, comprising administering to the individual an effective amount of the isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88. In some cases, such methods or uses comprise administering to the individual at least one other therapeutic agent, such as an immune checkpoint inhibitor, a chemotherapeutic agent, an anti-angiogenic agent, or an anti-tumor agent, or another therapy, such as radiation therapy, hormone therapy, or surgery.

[0151] In other embodiments, the antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88 can be used to prepare a medicament for treating an infectious disease, or the isolated antibody of any one of embodiments 1 to 87 or the pharmaceutical composition of embodiment 88 can be used to treat an infectious disease. Other embodiments include a method of treating an infectious disease in a subject in need thereof, comprising administering an effective amount of an antibody herein. In some such uses and methods herein, the antibody is administered together with another therapeutic agent.

[0152] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the claims. All references cited herein are incorporated herein by reference in their entirety. BRIEF DESCRIPTION OF THE DRAWINGS

[0153] Figures 1A to 1D Activity of exemplary murine anti-human PAD4 antibody (Ab) clones is shown. Figure 1A and Figure 1B The effect of antibody clones on PAD4 activity based on the conversion of arginine to citrulline was shown. Figure 1A Shows the percentage of PAD4 activity and Figure 1B Citrulline concentrations in the presence of antibody clones are shown. Figure 1C and Figure 1D The antibody was shown to inhibit PAD4 in a dose-dependent manner. Figure 1C Dose response curve for antibody clone 13 is shown. Figure 1D Dose response curves for antibody clone 20 are shown.

[0154] Figures 1E to 1H The VH and VL amino acid sequences of clone 13 are shown (VH= Figure 1E ; VL= Figure 1F ) and the VH and VL amino acid sequences of clone 20 (VH= Figure 1G ; VL= Figure 1H ).exist Figures 1E to 1HIn the sequence shown in , (1) HCDR1, HCDR2, and HCDR3 sequences are located at amino acid positions 26 to 35, 50 to 65, and 95 to 102, respectively; and (2) LCDR1, LCDR2, and LCDR3 sequences are located at amino acid positions 24 to 34, 50 to 56, and 89 to 97, respectively. Figure 1E and Figure 1G In the AbM definition of CDRs, HCDR1 is shown at amino acid positions 26 to 35 ( Figure 1E and Figure 1G (Abhinandan and Martin, Mol. Immunol. 45:3832-3839 (2008); Swindells et al., J. Mol. Biol. 429:356-364 (2017)). According to the Kabat numbering scheme, HCDR1 is actually located at amino acid positions 31 to 35 (not shown). The remaining CDRs (HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3; Figures 1E to 1H ) are numbered according to the Kabat numbering scheme.

[0155] Figure 2 It was shown that the PAD4 antibody was specific for PAD4 and did not substantially inhibit PAD2 activity.

[0156] Figures 3A to 3X Antibody stability was demonstrated, as assessed by size exclusion chromatography (SEC) and peptide mapping. Figure 3A SEC results are shown for 20 exemplary humanized (hz) clone 13 and 20 antibodies. Figure 3B Peptide mapping results for an exemplary hz clone 20 antibody are shown. Figures 3C to 3D Peptide mapping results for an exemplary hz clone 13 antibody are shown. Figure 3E and Figure 3F SEC results of hz13-12 and hz13-5 are shown separately. Figures 3G to 3J Peptide mapping results for hz13-12 are shown. Figures 3K to 3N Peptide mapping results for hz13-5 are shown. Figure 3O and Figure 3P SEC results of hz13-12 D31E and hz13-5 D31E are shown separately. Figure 3Q and Figure 3R The peptide mapping results of hz13-12 D31E and hz13-5 D31E are shown respectively. Figure 3S SEC results for hz13-5 are shown. Figure 3T SEC results for hz13-5 D31E are shown. Figure 3U and Figure 3V The peptide localization results of high concentration hz13-5 and hz13-5D31E are shown respectively. Figure 3W and Figure 3X The SEC results and peptide localization results of hz13-5 D31E are shown respectively.

[0157] Figures 4A to 4B Clone 13 ( Figure 4A ) and clone 20 ( Figure 4B ) epitopes. Regions in hPAD4 where HDX was significantly reduced upon Fab binding are boxed (black boxes indicate primary epitopes; gray boxes indicate secondary epitopes). For each antibody, the primary epitope is indicated by underlined black text above the corresponding black box, and the secondary epitope is indicated by smaller text above the corresponding gray box.

[0158] Figures 5A to 5B Clone 13 ( Figure 5A ) and clone 20 ( Figure 5B ) binding residues. Protected residues for which the difference in % labeling between hPAD4 and hPAD4 / Fab was statistically significant (p-value < 0.01 based on Student's t-test) are marked with an asterisk. Insufficient MS / MS fragmentation was obtained to distinguish between protection of H644 and C645; both residues are considered binding residues for clone 20.

[0159] Figure 6 An epitope summary is shown for clone 13 and clone 20. The epitope region determined by HDX-MS is boxed. The binding residues determined by FPOP, GEE, and DEPC labeling are underlined and highlighted in bold.

[0160] 7A to 7B Pepsin was shown to be effective for clone 13 (HC; Figure 7A ) and clone 13 light chain (LC; Figure 7B ). Each bar represents a pepsin peptide. The complementarity determining regions (CDRs) are boxed.

[0161] Figures 8A to 8B Clone 13 Fab HC ( Figure 8A ) and clone 13Fab LC ( Figure 8B ) The region with significantly reduced HDX. The paratope region of clone 13 is boxed.

[0162] Figures 9A to 9B The cryo-electron microscopy (Cryo-EM) structure of the PAD4.Clone 13 Fab complex is shown. Figure 9A The PAD4.Clone 13 Fab monomer is shown. PAD4 residues are numbered using a three-letter code. The approximate location of the catalytic site is circled. Figure 9B The PAD4.Fab13 dimer is shown.

[0163] Figure 10 Residue D31 of clone 13 and the interaction at the interface of PAD4 and the heavy chain of clone 13 Fab are shown. Fab residues are numbered with a single residue code, while PAD4 residues are numbered with a three-letter code.

[0164] Figure 11 The substrate binding site of human PAD4 (PDB code 2DEW) is shown. The lysine loop and catalytic helix are shown (the lysine loop includes LYS-519 and ILE-526, and the catalytic helix includes ILE-630 and ILE-638). The arginine residues of the cocrystal substrate peptide are shown in stick representation. 2+ Ions are shown as spheres.

[0165] Figure 12 A superposition of the PAD4 chains from the 2DEW and PAD.Clone 13 Fab cryoEM structures is shown. 2DEW is shown as a cartoon representation shaded in light grey, while PAD4 from the PAD4.Clone 13 Fab cryoEM structure is shown as a black cartoon. The heavy and light chains of Clone 13 Fab are shown as thin ribbons. PAD4 residues are numbered with a three-letter code, while Clone 13 Fab residues are numbered with a single-letter code. The movement of the lysine loop and its effect on the catalytic helix are shown by the black arrows. 2+ Ions are shown as spheres.

[0166] Figures 13A to 13N Heatmap showing pH-dependent anti-PAD4 antibody studies ( Figures 13A to 13L ) and SPR( Figures 13M to 13N ) measured value. Figures 13A to 13L In the heat map shown, (1) ER values ​​less than 1 indicate that the substitution will weaken binding to PAD4, (2) values ​​greater than 1.00 indicate that the amino acid substitution will result in stronger binding to PAD4, and (3) 1.00 indicates that the amino acid substitution will not result in a change in binding (i.e., binding will not be stronger or weaker due to the mutation). At pH 7 ( Figure 13A 、 Figure 13C 、 Figure 13E 、 Figure 13G 、 Figure 13I and Figure 13K ) and pH 6( Figure 13B 、 Figure 13D 、 Figure 13F 、 Figure 13H 、 Figure 13J and Figure 13L ) were used to test pH-dependent anti-PAD4 antibodies. Figure 13A HCDR1 of clone 13 (mAb13) at pH 7 is shown. Figure 13BHCDR1 of clone 13 (mAb13) at pH 6 is shown. Figure 13C HCDR2 of clone 13 (mAb13) at pH 7 is shown. Figure 13D HCDR2 of clone 13 (mAb13) at pH 6 is shown. Figure 13E HCDR3 of clone (mAb13) at pH 7 is shown. Figure 13F HCDR3 of clone 13 (mAb13) at pH 6 is shown. Figure 13G LCDR1 of clone 13 (mAb13) at pH 7 is shown. Figure 13H LCDR1 of clone 13 (mAb13) at pH 6 is shown. Figure 13I LCDR2 of clone 13 (mAb13) at pH 7 is shown. Figure 13J LCDR2 of clone 13 (mAb13) at pH 6 is shown. Figure 13K HCDR3 of clone 13 (mAb13) at pH 7 is shown. Figure 13L LCDR3 of clone 13 (mAb13) at pH 6 is shown. Figures 13M to 13N The exemplary anti-PAD4 antibody hz13-5 was shown to be effective at pH 7.6 ( Figure 13M ) and pH 6.0 ( Figure 13N ). The pH-dependent variant * is hz13-5 VH_D31H::Vk_I30H. A slight increase in the dissociation phase signal was observed at pH 6.0, which may be due to the lower stability of PAD4 at pH 6.0, which may lead to some aggregation of PAD4 on the surface.

[0167] Figure 14 Shown are the percentages of healthy volunteer human donors (n=40) with positive CD4+ proliferation responses after 7 days of incubation of autologous antigen-pulsed monocyte-derived DCs with the antibody Avastin, IL-21R mAb (ATR-107), clone 13-based hz13-12 antibody, clone 20-based hz20-7, clone 13-based hz13-5, and clone 13-based hz13-5 D31E.

[0168] Figure 15 Dose-dependent inhibition of human PAD4 with antibody hz13-5D31E is shown at four concentrations of recombinant human PAD4 (rhPAD4) ranging from 13.5 nM (1 μg / mL) to 108 nM (8 μg / mL).

[0169] 16A to 16FShown is the reduction of extracellular citrullinated histone 3 in human CD14+ monocytes stimulated with lipopolysaccharide (LPS) at the indicated concentrations of the indicated clone 13 (C13) or clone 20 (C20) antibody variants or treated with an isotype control antibody. Figure 16A Shown are the citrullinated histone 3 levels in mononuclear spheres incubated with hz13-5. Figure 16B Shown are the citrullinated histone 3 levels in mononuclear spheres incubated with hz13-5D31E. Figure 16C Shown are the citrullinated histone 3 levels in mononuclear spheres incubated with hz20-2. Figure 16D Shown is the content of citrullinated histone 3 in mononuclear spheres incubated with hz20-7. Figure 16E Shown are the citrullinated histone 3 levels in mononuclear spheres incubated with hz13-3. Figure 16F Shown are the citrullinated histone 3 levels in mononuclear spheres incubated with hz13-12.

[0170] 17A to 17F Shown is the reduction of GM-CSF (ng / mL) in human CD14+ monocytes stimulated with LPS at the indicated concentrations of the indicated clone 13 (C13) or clone 20 (C20) antibody variants or treated with an isotype control antibody. Figure 17A Shown is the detection of GM-CSF in monocytes incubated with hz13-5. Figure 17B The GM-CSF content in monocytes incubated with hz13-5 D31E is shown. Figure 17C The GM-CSF content in monocytes incubated with hz13-3 is shown. Figure 17D The GM-CSF content in monocytes incubated with hz13-12 is shown. Figure 17E The GM-CSF content in monocytes incubated with hz20-2 is shown. Figure 17F The GM-CSF content in monocytes incubated with hz20-7 is shown.

[0171] 18A to 18B Shown is the reduction in GM-CSF gene expression in LPS-stimulated human CD14+ monocytes treated with the indicated concentrations of the indicated clone 13 (C13) antibody variants (fold induction, calculated based on 2,-deltadeltaCT). Figure 18A Shown are the fold induction of GM-CSF in monocytes incubated with hz13-5. Figure 18BFold induction of GM-CSF in mononuclear cells incubated with hz13-5 D31E is shown. GM-CSF gene Ct values ​​were normalized to the housekeeping gene peptidylprolylcis-transisomerase A (PPIA) by subtracting the Ct values ​​from the PPIA to generate the DCt. The relative quantity (RQ) value (fold induction) of each sample for the housekeeping gene was calculated using the following formula: RQ = 2 -DCt .

[0172] Figures 19A to 19B Detection of the anti-PAD4 antibody hz13-5D31E in the acidic cellular compartment is shown. Figure 19A Shown are images of donor GAC-077 stained red with Fabfluor-pH (left panel) and an overlay of FabFluor-pH and Cytotox green staining (center and right panels; co-staining appears yellow) at 48 h under the indicated conditions (center panel = 50 μg / mL hz13-5 D31E; right panel = 50 μg / mL isotype control). Figure 19B The antibody internalization area (μm) measured by FAbFluor-pH staining under the conditions of two donors GAC-077 and GAC-045 is shown. 2 / image).

[0173] Figure 20 In vivo efficacy of exemplary anti-PAD4 antibodies, anti-murine PAD4 mAbs, and mumAbs in the lipopolysaccharide (LPS)-induced acute pulmonary inflammation pharmacodynamics (ALIPD) model is shown. 30 mpk and 100 mpk treatments resulted in 74% and 83% inhibition of extracellular citrullinated H3, respectively, compared to isotype control (100 mpk) conditions. "Mpk" = mg / Kg. "IC" = isotype control antibody. "Untreated ” = without LPS induction.

[0174] Figures 21A to 21B Shows the experimental design for the acute joint inflammation (AJI) and chronic joint inflammation (CJI) studies. Figure 21A The schedule for treatment administration (subcutaneous anti-PAD4 antibody or isotype control), LPS administration to induce inflammation, and outcome assessment is shown. Figure 21B Procedures for displaying induction and evaluating results.

[0175] FIG. 22A to FIG. 22B We show that extracellular citrullinated proteins are reduced by an anti-murine PAD4 mAb (mumAb) in an LPS-induced acute joint inflammation (AJI) pharmacodynamic (PD) model. Figure 22A Displaying Cit-ITIH4 results. Figure 22B Show Cit-PRG4 results. Figure 20 The percentage inhibition was calculated in a similar manner as described. "Mpk" = mg / kg. "Isotype" = isotype control antibody (IC). "Untreated" = no LPS injection.

[0176] FIG. 23A to FIG. 23B We show that extracellular citrullinated proteins are reduced by an anti-murine PAD4 mAb (mumAb) in an LPS-induced chronic joint inflammation (CJI) pharmacodynamic (PD) model. Figure 23A Displaying Cit-ITIH4 results. Figure 23B Show Cit-PRG4 results. Figure 20 Percent inhibition was calculated in a similar manner as described. "Mpk" = mg / kg. "Isotype" = isotype control antibody (IC). "Untreated" = no LPS injection. "PBS" = untreated mice sham-injected with phosphate-buffered saline (PBS) instead of LPS.

[0177] FIG. 24A to FIG. 24B We show that anti-PAD4 mAb (mumAb) reduces NETosis in a pristane-induced peritonitis model. Figure 24A ) and METosis( Figure 24B N = 6 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to isotype control by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = sham injection of PBS instead of LPS.

[0178] FIG. 25A to FIG. 25B We show that Cit-H3 in neutrophils and monocytes is reduced by anti-murine PAD4 mAb (mumAb) in a pristane-induced peritonitis model. Figure 25A Shows cell % and Figure 25B Cell numbers are shown. N = 6 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to isotype control by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = did not receive pristane injection.

[0179] Figures 26A to 26B showed that anti-murine PAD4 mAb (mumAb) reduced M1 macrophages ( Figure 26A ) and M2 macrophages ( Figure 26B) in Cit-H3. N = 6 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to isotype control by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = not induced with pristane.

[0180] FIG. 27A to FIG. 27B showed that anti-murine PAD4 mAb (mumAb) reduced elastase in peritoneal fluid samples in a pristane-induced peritonitis model ( Figure 27A ) and MPO( Figure 27B N = 6 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared to isotype control by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = not induced with pristane.

[0181] FIG. 28A to FIG. 28B We show that chemokines and cytokines are reduced by anti-murine PAD4 mAb (mumAb) in a pristane-induced peritonitis model. Figure 28A The contents of GROβ / MIP-2α, GROα / KC, MCP 1 and MIP 1β in peritoneal fluid are shown. Figure 28B Plasma IL-6 and MIP3a levels are shown. N = 6 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared to isotype control by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = not induced with pristane.

[0182] Figures 29A to 29C Results of a preventive study in a collagen-induced arthritis (CIA) model are shown. Anti-murine PAD4 mAb (mumAb) reduced clinical scores ( Figure 29A ), disease incidence ( Figure 29B ) and paw weight ( Figure 29C N = 10 to 15 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "IC" = isotype control antibody. "Untreated" = no collagen induction.

[0183] Figures 30A to 30E Shown are the results of a preventive study in a CIA model. Anti-murine PAD4 mAb (mumAb) reduced immune cell infiltration in the paw. Figure 30A :CD45+ cells. Figure 30B : Total bone marrow cells; CD11b+ cells. Figure 30C : Neutrophils; CD11b+Ly6G+Ly6C- cells. Figure 30D : Monocytes; CD11b+Ly6G-Ly6C+ cells. Figure 30E = Macrophages; F4 / 80+ cells. N = 10 to 15 cells per treatment group. N = 5 cells in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = no collagen induction.

[0184] Figures 31A to 31B Results from a preventive study in a CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduces NETosis in mouse paws ( Figure 31A ) and METosis( Figure 31B (left panel shows monocyte counts, and right panel shows macrophage counts). N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = no collagen induction.

[0185] Figures 32A to 32C Results of a preventive study in a CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduced neutrophil counts in the paws of mice ( Figure 32A ), single core ball ( Figure 32B ) and M2 macrophages ( Figure 32C ) of total Cit-H3 in each treatment group. N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC).

[0186] Figures 33A to 33C Results from a preventive study in a CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduced PAD4 expression in the paws of mice ( Figure 33A )、MPO( Figure 33B ) and neutrophil elastase ( Figure 33CN = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "Untreated" = no collagen induction.

[0187] Figures 34A to 34B Results from a preventive study in a CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduced anti-citrullinated peptide antibodies (ACPAs) in paws and serum. Figure 34A ), and also reduced serum anti-CII antibody titer ( Figure 34B N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "Untreated" = no collagen induction.

[0188] Figure 35 Results from a preventive study in a CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduced paw histology scores (% reduction presented is relative to the IC). N = 10 to 15 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "IC" = isotype control antibody.

[0189] Figures 36A to 36C Results from a semi-therapeutic study in a CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduced clinical scores ( Figure 36A ), disease incidence ( Figure 36B ) and paw weight ( Figure 36C N = 10 to 15 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "IC" = isotype control antibody.

[0190] Figures 37A to 37E Shown are the results of a semi-therapeutic study in a CIA model. Anti-murine PAD4 mAb (mumAb) reduces immune cell infiltration in the paw. Figure 37A : Total cell infiltration; CD45+ cells. Figure 37B : Total bone marrow cells; CD11b+ cells. Figure 37C : Neutrophils; CD11b+Ly6G+Ly6C- cells. Figure 37D : Monocytes; Ly6C+ cells. Figure 37E= Macrophages; F4 / 80+ cells. N = 10 to 15 cells per treatment group. N = 5 cells in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = no collagen induction.

[0191] Figures 38A to 38B Results from a semi-therapeutic study in a CIA model are presented. Anti-murine PAD4 mAb (mumAb) reduces NETosis in mouse paws ( Figure 38A ) and METosis( Figure 38B N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to IC by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = no collagen induction.

[0192] Figures 39A to 39C Results from a semi-therapeutic study in a CIA model are presented. Anti-murine PAD4 mAb (mumAb) reduced neutrophil counts in the paws of mice ( Figure 39A ), single core ball ( Figure 39B ) and M2 macrophages ( Figure 39C ) in total Cit-H3. N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "ISO" = isotype control antibody (IC). "Untreated" = no collagen induction.

[0193] Figures 40A to 40C Results from a semi-therapeutic study in a CIA model are presented. Anti-murine PAD4 mAb (mumAb) reduced PAD4 expression in mouse paws ( Figure 40A )、MPO( Figure 40B ) and neutrophil elastase ( Figure 40C N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "Untreated" = no collagen induction.

[0194] Figures 41A to 41BResults of a semi-therapeutic study in a CIA model are presented. Anti-murine PAD4 mAb (mumAb) reduced ACPA levels in paws and serum ( Figure 41A , right and left figures respectively) and serum anti-CII antibody titers ( Figure 41B N = 10 to 15 per treatment group. N = 5 in the untreated group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg.

[0195] Figure 42 Results from a semi-therapeutic study in the CIA model are shown. Anti-murine PAD4 mAb (mumAb) reduced paw histological scores. N = 10 to 15 per treatment group. *p < 0.05, **p < 0.01, ***p < 0.001 compared to vehicle by one-way ANOVA with Dunnett's test. "Mpk" = mg / kg. "IC" = isotype control antibody.

[0196] Figures 43A to 43E Demonstrates efficacy of anti-human PAD4 mAb clone 20 and humanized derivatives in the LPS ALIPD model. Figure 43A 、 Figure 43B and Figure 43C Shown are the effects of clone 20, humanized clone 20-based hz20-2 antibody, and humanized clone 20-based hz20-7 antibody, respectively, at the indicated doses on extracellular citrullinated H3 relative to total extracellular H3; percent reduction represents reduction relative to isotype control conditions. Figure 43D and Figure 43E Shown are the effects of the clone 20-based hz20-2 and clone 20-based hz20-7 antibodies, respectively, at the indicated doses on extracellular citrullinated ITIH4 relative to total extracellular ITIH4; the percent reduction represents the reduction relative to the isotype control condition. "Mpk" = mg / kg. "IC" = isotype control antibody. "Untreated" = not nebulized with LPS.

[0197] Figures 44A to 44E Demonstrated efficacy of anti-human PAD4 mAb clone 13 and humanized derivatives in the LPS ALIPD model. Figure 44A 、 Figure 44B and Figure 44C Shown are the effects of clone 13, humanized clone 13-based hz13-12 antibody, and humanized clone 13-based hz13-5 antibody, respectively, on extracellular citrullinated H3 relative to total extracellular H3 at the indicated doses; percent reduction represents reduction relative to isotype control conditions. Figure 44D and Figure 44EShown are the effects of the clone 13-based hz13-12 antibody and the clone 13-based hz13-5 antibody, respectively, at the indicated doses on extracellular citrullinated ITIH4 relative to total extracellular ITIH4; the percent reduction represents the reduction relative to the isotype control condition. "Mpk" = mg / kg. "IC" = isotype control antibody. "Untreated" = not nebulized with LPS.

[0198] Figures 45A to 45E Shown is the experimental design of the LPS AJIPD study using Hu-PAD4 knock-in mice and human PAD4 endpoints. Figure 45A Displays the research timeline. Figure 45B An overview of the experimental procedures is shown, including treatment at the knee joint and extraction of citrullinated proteins from explanted tissue. Figure 45C ), extracellular Cit-ITIH4 ( Figure 45D ) and human PAD4( Figure 45E ).

[0199] Figures 46A to 46C Shows the efficacy of anti-human PAD4 antibodies in LPS AJIPD studies. Shows the hz13-12 antibody based on clone 13 at the indicated doses ( Figure 46A ), hz13-5 antibody based on clone 13 ( Figure 46B ) and hz20-2 antibody based on clone 20 ( Figure 46C ) Effect on extracellular citrullinated ITIH4 relative to total extracellular ITIH4; percent reduction represents the reduction relative to isotype control conditions. "Mpk" = mg / kg. "IC" = isotype control antibody. "Untreated" = no pristane injection. "PBS" = no treatment, injection of phosphate-buffered saline instead of LPS.

[0200] Figures 47A to 47C Shows the efficacy of anti-human PAD4 antibody in LPS AJIPD study. Shows extracellular Cit-PRG4 in patellar explant supernatants. Shows the hz13-12 antibody based on clone 13 at the indicated doses ( Figure 47A ), hz13-5 antibody based on clone 13 ( Figure 47B ) and hz20-2 antibody based on clone 20 ( Figure 47C ) Effect on extracellular citrullinated PRG4 relative to total extracellular PRG4; percent reduction represents the reduction relative to isotype control conditions. "Mpk" = mg / kg. "IC" = isotype control antibody. "PBS" = phosphate-buffered saline. "Untreated" = no LPS injection.

[0201] Figures 48A to 48BThe efficacy of anti-human PAD4 antibodies hz13-5 and hz13-5D31E in LPS AJI studies was demonstrated. These antibodies inhibit ITIH4 ( Figure 48A ) and PRG4( Figure 48B ) citrullination; the results show that hz13-5 D31E antibody has a dose-dependent inhibitory effect. "Mpk" = mg / kg. "IC" = isotype control antibody. "Untreated" = no LPS injection.

[0202] Figures 49A to 49B The hz13-5D31E antibody was shown to maintain efficacy in the presence of endogenous PAD4 antibodies from RA patients. Figure 49A Shown are PAD4 autoantibodies measured by OD450 in ELISA from purified IgG of serum samples from 21 RA patients and 10 healthy control individuals (NHV). Figure 49B Inhibition of H3 citrullination by hz13-5D31E in the presence of purified IgG from RA (filled squares) or NHV (open squares) serum is shown. Dashed lines indicate citrullination of H3 by PAD4 in the presence of purified IgG from RA donors (lower line) and NHV donors (upper line) in the absence of hz13-5 D31E. DETAILED DESCRIPTION

[0203] I. Definition

[0204] Unless otherwise defined, scientific and technical terms used in conjunction with the present invention will have the meanings commonly understood by those skilled in the art.In addition, unless the context requires otherwise, singular terms shall include pluralities and plural terms shall include the singular.

[0205] In this application, the use of "or" means "and / or" unless otherwise stated. In the context of multiple dependent items, the use of "or" only refers to more than one of the preceding independent or dependent items in an alternative manner. In addition, unless specifically stated otherwise, terms such as "element" or "component" encompass both elements and components comprising one unit and elements and components comprising more than one subunit.

[0206] Exemplary techniques used in conjunction with recombinant DNA, oligonucleotide synthesis, tissue culture and transformation (e.g., electroporation, lipofection), enzymatic reactions, and purification techniques are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989)).

[0207] As used herein, the term "about" refers to a numerical value, including, for example, integers, fractions, and percentages, whether or not explicitly indicated. The term "about" generally refers to a range of numerical values ​​(e.g., + / - 5-10% of the listed range) that a person of ordinary skill in the art would consider to be equal to the listed value (e.g., having the same function or result). When terms such as at least and approximately appear before a list of numerical values ​​or ranges, the terms will modify all values ​​or ranges provided in the list. In some cases, the term approximately may include numerical values ​​rounded to the nearest significant figure.

[0208] The term "polypeptide" refers to a polymer of amino acid residues and is not limited to a minimum length. A "protein" may comprise one or more polypeptides. Such polymers of amino acid residues may contain natural or non-natural amino acid residues and include, but are not limited to, peptides, oligopeptides, dimers, trimers, and multimers of amino acid residues. The definition encompasses full-length proteins and fragments thereof. The term also includes post-expression modifications of the polypeptide, such as glycosylation, sialylation, acetylation, phosphorylation, and the like. Furthermore, for the purposes of the present invention, "polypeptide" or "protein" refers to a polypeptide or protein, respectively, that includes modifications of the native sequence, such as deletions, additions, and substitutions (generally conservative in nature), as long as the protein maintains the desired activity. These modifications may be intentional, such as through site-directed mutagenesis, or may be accidental, such as through mutations in the host producing the protein or errors due to PCR amplification. A protein may comprise two or more polypeptides.

[0209] As used herein, "PAD4" or "protein arginine deiminase 4" or "peptidyl arginine deiminase 4" refers to human PAD4 (huPAD4; UniProt ID: Q9UM07), unless otherwise specified (i.e., murine PAD4, macaque PAD4, or the like). Exemplary human PAD4 amino acid sequences are shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.

[0210] The term "antibody" herein refers to a molecule comprising at least complementarity determining regions (CDRs) 1, 2, and 3 of a heavy chain and at least CDRs 1, 2, and 3 of a light chain, wherein the molecule is capable of binding to an antigen. The term is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies, diabodies, etc.), full-length antibodies, single-chain antibodies, antibody conjugates, and antibody fragments, as long as they exhibit the desired PAD4-specific binding activity.

[0211] An "isolated" antibody is one that has been separated from the components of its natural environment. In some aspects, the antibody is purified to a purity greater than 95% or 99% as determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[0212] "Antigen" refers to the antibody target, that is, the molecule to which the antibody specifically binds. The term "epitope" refers to the site on a protein or non-protein antigen to which the antibody binds. Epitopes on proteins can be formed by continuous stretches of amino acids (linear epitopes) or comprise non-contiguous amino acids that are spatially close, for example, due to the folding of the antigen (i.e., by the tertiary folding of the protein antigen) (conformational epitopes). Linear epitopes typically remain bound to the antibody after the protein antigen is exposed to a denaturant, while conformational epitopes are typically destroyed after treatment with a denaturant.

[0213] "Anti-PAD4 antibody" or "PAD4 antibody" or "antibody that specifically binds to PAD4" or "antibody that binds to PAD4" and similar phrases refer to an antibody that specifically binds to PAD4 as defined herein.

[0214] The term "heavy chain" refers to a polypeptide comprising at least one heavy chain variable region, with or without a leader sequence. In some embodiments, a heavy chain comprises at least a portion of a heavy chain constant region. The term "full-length heavy chain" refers to a polypeptide comprising a heavy chain variable region and a heavy chain constant region, with or without a leader sequence.

[0215] The term "light chain" refers to a polypeptide comprising at least one light chain variable region, with or without a leader sequence. In some embodiments, a light chain comprises at least a portion of a light chain constant region. The term "full-length light chain" refers to a polypeptide comprising a light chain variable region and a light chain constant region, with or without a leader sequence.

[0216] As used herein, the term "complementarity determining region" ("CDR") refers to each region of the variable region of an antibody whose sequence is highly variable and determines antigen binding specificity. Generally speaking, an antibody comprises six CDRs: three located in VH (CDR-H1 or heavy chain CDR1, CDR-H2, CDR-H3) and three located in VL (CDR-L1, CDR-L2, CDR-L3). Unless otherwise indicated, CDRs are identified according to the sequence listing herein.

[0217] "Framework" or "FR" refers to variable region residues that are not part of the complementarity determining regions (CDRs). The variable region FRs generally consist of four FRs: FR1, FR2, FR3, and FR4. Thus, the CDR and FR sequences typically appear in the following order in VH (or VL): FR1-CDR-H1 (CDR-L1)-FR2-CDR-H2 (CDR-L2)-FR3-CDR-H3 (CDR-L3)-FR4.

[0218] The term "variable region" or "variable domain" refers to the antibody heavy chain or light chain domain involved in the binding of the antibody to the antigen. The variable domains of the heavy and light chains of natural antibodies (VH and VL, respectively) generally have similar structures, each domain comprising four conserved framework regions (FR) and three complementarity determining regions (CDRs). See, for example, Kindt et al., Kuby Immunology, 6th edition, WH Freeman and Co., page 91 (2007). The variable domain may comprise a heavy chain (HC) CDR1-FR2-CDR2-FR3-CDR3, with or without all or a portion of FR1 and / or FR4; and a light chain (LC) CDR1-FR2-CDR2-FR3-CDR3, with or without all or a portion of FR1 and / or FR4. That is, the variable domain may lack a portion of FR1 and / or FR4, as long as it retains antigen binding activity. A single VH or VL domain may be sufficient to confer antigen binding specificity. In addition, antibodies that bind to a specific antigen can be separated from antibodies that bind to the antigen using a VH or VL domain to screen a collection of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0219] The "constant region" of the light and heavy chains of an antibody refers to the sequence portions other than the FRs and CDRs and the variable regions. Certain antibody fragments may lack all or some of the constant regions. Each heavy chain has a variable domain (VH) (also referred to as a variable heavy domain or heavy chain variable region) from N-terminus to C-terminus, followed by three constant heavy domains (CH1, CH2, and CH3). Similarly, each light chain has a variable domain (VL) (also referred to as a variable light domain or light chain variable region) from N-terminus to C-terminus, followed by a constant light domain (CL).

[0220] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In one aspect, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to Gly446 and Lys447 (EU numbering) at the carboxyl terminus of the heavy chain. The antibody produced by the host cell may undergo post-translational cleavage of one or more (especially one or two) amino acids at the C-terminus of the heavy chain. Therefore, the antibody produced by the host cell by expressing a specific nucleic acid molecule encoding a full-length heavy chain may include a full-length heavy chain, or it may include a split variant of the full-length heavy chain. It may be the case that the last two C-terminal amino acids of the heavy chain are glycine and lysine, respectively. Therefore, the C-terminal lysine or the C-terminal glycine and lysine in the Fc region may be present or absent. Therefore, a "full-length heavy chain constant region" or "full-length antibody" is, for example, a human IgG1 antibody, including an IgG1 having both a C-terminal glycine and a lysine, but not containing a C-terminal lysine or not containing both a C-terminal glycine and a lysine. Unless otherwise indicated herein, amino acid residues in the Fc region or constant region are numbered according to the EU numbering system (also referred to as the EU index) as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0221] "Effector functions" refer to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation.

[0222] The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these antibodies can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The light chains of antibodies can be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of their constant domains.

[0223] An "antibody fragment" is a molecule other than an intact antibody that comprises a portion of an intact antibody that binds to the antigen bound by the intact antibody (i.e., PAD4). Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, and F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv and scFab); single-domain antibodies (dAbs); and multispecific antibodies formed from antibody fragments. For a review of certain antibody fragments, see Holliger and Hudson, Nature Biotechnology 23:1126-1136 (2005).

[0224] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody that has a structure substantially similar to a native antibody structure, or in the case of an IgG antibody, has heavy chains that contain an Fc region as defined above.

[0225] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0226] A "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human CDRs and amino acid residues from human FRs. In certain aspects, a humanized antibody will comprise substantially all of at least one and typically two variable domains, wherein all or substantially all of the CDRs correspond to the CDRs of a non-human antibody, and all or substantially all of the FRs correspond to the FRs of a human antibody. A humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody (e.g., a non-human antibody) refers to an antibody that has been humanized.

[0227] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, that is, except for example containing naturally occurring mutations or possible variant antibodies produced during the preparation of a monoclonal antibody preparation, the individual antibodies constituting the population are identical and / or bind to the same epitope, and the variants are generally present in small amounts. In contrast to polyclonal antibody preparations that typically include different antibodies for different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is for a single determinant on an antigen. Therefore, the modifier "monoclonal" represents the antibody properties obtained from a substantially homogeneous antibody population, and should not be interpreted as requiring antibodies to be produced by any ad hoc method.

[0228] "Multispecific" antibodies are antibodies that specifically bind to more than one target antigen, while "bispecific" antibodies are antibodies that specifically bind to two antigens. "Antibody conjugates" are antibodies conjugated to one or more heterologous molecules, including but not limited to therapeutic agents or labels.

[0229] Antibodies can be modified as part of certain host cell production processes or through in vivo metabolism. The antibody or antibody region amino acid sequences herein are intended to encompass not only the specific amino acid sequence, but also post-translationally modified sequences, including, for example, side chain modifications and cleavage. Such post-translational modifications may occur, for example, as a result of producing the antibody in a host cell and / or as a result of in vivo post-translational modifications in an animal (e.g., a human).

[0230] In some embodiments, the antibodies disclosed herein comprise post-translational modifications (e.g., one or more post-translational modifications). Post-translational modifications may include, for example, ubiquitination, phosphorylation, acetylation, hydroxylation, methylation, glycosylation, monophosphate adenylation, prenylation, deamidation, elimination (elimylation), citrullination, and carbamylation. In some embodiments, the antibodies are not post-translationally modified.

[0231] As noted above, the antibody may undergo post-translational cleavage of one or more, particularly one or two, amino acids from the C-terminus of the heavy chain (typically Gly-Lys). This cleavage may occur, for example, due to the process of producing the antibody in a host cell. The antibody produced by expressing a specific nucleic acid molecule encoding a full-length heavy chain may comprise a full-length heavy chain, or it may comprise a cleavage variant of the full-length heavy chain, such as a heavy chain lacking a C-terminal Lys or a C-terminal Lys.

[0232] Other types of post-translational modifications may occur during antibody production or otherwise in vivo, such as modifications of amino acid side chains. For example, the N-terminal Glu or Gln residue on the antibody chain may be post-translationally modified to N-terminal pyroglutamate (also known as pyrrolidine carboxylate; abbreviated as pE).

[0233] "Percent (%) amino acid sequence identity" and "homology" with respect to peptide, polypeptide or antibody sequences are defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular peptide or polypeptide sequence, after aligning the sequences and, if necessary, introducing gaps to achieve the maximum percentage of sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be achieved in various ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or MEGALIGN™ (DNASTAR) software. Appropriate parameters for measuring alignment can be determined by one skilled in the art, including any algorithm required to achieve maximum alignment over the full length of the compared sequences.

[0234] The term "signal sequence" or "leader sequence" refers to a sequence of amino acid residues located at the N-terminus of a polypeptide that facilitates secretion of the polypeptide from mammalian cells. The leader sequence can be cleaved to form a mature protein when the polypeptide is exported from the mammalian cell. The leader sequence can be natural or synthetic and can be heterologous or homologous to the protein to which it is attached. Non-limiting exemplary leader sequences also include leader sequences from heterologous proteins. In some embodiments, the antibody lacks a leader sequence. In some embodiments, the antibody comprises at least one leader sequence, which can be selected from a natural antibody leader sequence and a heterologous leader sequence.

[0235] The term "nucleic acid molecule" or "polynucleotide" includes any compound and / or substance comprising a nucleotide polymer. Each nucleotide is composed of a base, specifically a purine or pyrimidine base (i.e., cytosine (I), guanine (G), adenine (A), thymine (T), or uracil (U)), a sugar (i.e., deoxyribose or ribose), and a phosphate group. Typically, a nucleic acid molecule is described by a base sequence, wherein the bases represent the primary structure (linear structure) of the nucleic acid molecule. The base sequence is typically represented from 5' to 3'. As used herein, the term nucleic acid molecule encompasses deoxyribonucleic acid (DNA), including, for example, complementary DNA (cDNA) and genomic DNA; ribonucleic acid (RNA), especially messenger RNA (mRNA); synthetic forms of DNA or RNA; and mixed polymers comprising two or more of these molecules. Nucleic acid molecules can be linear or circular. In addition, the term nucleic acid molecule includes both sense and antisense strands, as well as single-stranded and double-stranded forms. In addition, the nucleic acid molecules described herein may contain naturally occurring or non-naturally occurring nucleotides. Examples of non-naturally occurring nucleotides include modified nucleotide bases having derivatized sugar or phosphate backbone bonds or chemically modified residues. Nucleic acid molecules also encompass DNA and RNA molecules, which are suitable for use as vectors for direct expression of the antibodies of the invention in vitro and / or in vivo, for example, in a host or patient. Such DNA (e.g., cDNA) or RNA (e.g., mRNA, circular RNA) vectors may be unmodified or modified.

[0236] An "isolated" nucleic acid is one that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0237] "Isolated nucleic acid encoding an anti-PAD4 antibody" refers to one or more nucleic acid molecules encoding the anti-PAD4 antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules in a single vector or different vectors, and such nucleic acid molecules present at one or more locations in a host cell.

[0238] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of a host cell into which they are introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. These vectors are referred to herein as "expression vectors."

[0239] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells.

[0240] In the present invention, "binds," "binding," "specific binding," and similar terms, when referring to, for example, a protein and its ligand, an antibody and its antigenic target, or some other binding pair, mean that the binding affinity between the members of the binding pair is sufficiently strong that the interaction cannot be attributed to random molecular association (i.e., "non-specific binding"). Such binding typically requires a dissociation constant (K) of 1 μM or less. D ), and can typically involve K values ​​of 100 nM or less D .

[0241] "Affinity" refers to the sum of non-covalent interaction forces between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). Affinity can generally be expressed as the dissociation constant (K D The affinity of an antibody for an antigen can be measured by common methods known in the art, such as surface plasmon resonance (SPR).

[0242] As used herein, the term "agonist" refers to a substance, such as an antibody, that increases at least one activity or function of a molecule to which it binds, or otherwise activates or helps activate the molecule. As used herein, the term "antagonist" or "inhibitor" refers to a substance, such as an antibody, that decreases at least one activity or function of a molecule to which it binds, or otherwise blocks or inhibits at least one activity or function of the molecule.

[0243] The terms "inhibition" and "inhibit" more generally refer to reducing or stopping any event (such as protein ligand binding), or reducing or stopping any phenotypic characteristic, or reducing or stopping the incidence, extent or likelihood of that characteristic. "Reduce" or "inhibit" refers to reducing, lowering or suppressing an activity, function and / or amount as compared to a reference. The inhibition or reduction is not necessarily complete. For example, in certain embodiments, "reduce" or "inhibit" means being able to reduce overall by 20% or more. In another embodiment, "reduce" or "inhibit" means being able to reduce overall by 50% or more. In another embodiment, "reduce" or "inhibit" means being able to reduce overall by 75%, 85%, 90%, 95% or more.

[0244] As used herein, "treatment" encompasses any administration or administration of a therapeutic agent for a human disease, and includes inhibiting a disease or the progression of a disease or one or more symptoms of a disease, inhibiting or slowing a disease or its progression or one or more symptoms thereof, arresting its development, partially or completely relieving a disease or one or more symptoms thereof, or preventing recurrence of one or more symptoms of a disease.

[0245] The terms "subject" and "patient" are used interchangeably herein to refer to a human, unless specifically indicated otherwise (ie, a murine subject or the like).

[0246] As used herein, "autoimmune disease" or "autoimmune disease" encompasses diseases characterized by an individual's immune system attacking its own normal cells and tissues, and also encompasses immune-mediated diseases that may or may not be characterized by the presence of autoantibodies. Many non-limiting examples of autoimmune diseases are provided throughout the present invention. Some non-limiting examples of autoimmune diseases include rheumatoid arthritis (RA), lupus (e.g., systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g., ANCA-associated vasculitis), thrombosis (e.g., venous thrombosis), and inflammatory bowel disease (IBD) (e.g., ulcerative colitis, Crohn's disease). The terms "disease" and "disorder" are used interchangeably herein. In some embodiments, an autoimmune disease is characterized by the presence of autoantibodies.

[0247] The term "effective amount" or "therapeutically effective amount" refers to an amount of a drug that is effective to treat a disease or condition in a subject so as to partially or completely alleviate one or more symptoms. In some embodiments, an effective amount refers to an amount effective to achieve the desired therapeutic or preventive result at the desired dosage and for the desired period of time.

[0248] As used herein, a "biological sample" refers to a sample obtained from an individual or an animal. Examples of biological samples include tissue samples and liquid biological samples, such as whole blood, serum, plasma, blood supernatant, or synovial fluid. A biological sample can be obtained directly from an individual, or can first be chemically or physically modified in some manner prior to use, for example, to facilitate analysis of the sample.

[0249] II. Exemplary Anti-PAD4 Antibodies

[0250] Provided herein are antibodies that specifically bind to protein arginine deiminase 4 (PAD4). In some embodiments, the antibody inhibits an activity of PAD4, such as citrullination of arginine.

[0251] Clone 13 and related antibodies

[0252] For example, the disclosure herein relates to a panel of antibodies based on a murine anti-human antibody designated "Clone 13," as further described, for example, in Example 1 below. For example, Clone 13 was prepared in its original murine anti-human form and then humanized to generate a series of antibodies designated hz13-1 through hz13-12, of which hz13-5 and hz13-12 were further modified at positions D31 to D31E (antibodies hz13-5 D31E and hz13-12 D31E), as described in the Examples herein. Cryo-EM studies of the Clone 13 Fab binding to PAD4 were performed, and paratope mapping was performed to identify portions of the Clone 13 variable region and its humanized variants that directly contact PAD4, providing additional structural information regarding portions of the Clone 13 variable region and its related humanized antibodies that determine PAD4 binding. (See Examples 12-13 below.) Antibody hz13-5 was also further modified to identify antibodies that bind to PAD4 in a pH-dependent manner. (See the following subsections and Example 14).

[0253] For example, in some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 9.

[0254] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 62, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 9.

[0255] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4 or 62 and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7. In particular, paratope mapping and structural analysis of antibodies comprising the above heavy chain and light chain CDR sets revealed that HCDR1, HCDR3, and LCDR1 are in contact with PAD4. (See Figures 8A to 8B ). Thus, in some embodiments, the antibodies herein comprise the above-described three CDR sets. In some embodiments, the antibodies comprise a VH and a VL, wherein the VH comprises: a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4 and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and the VL comprises: a LCDR1 comprising the amino acid sequence of SEQ ID NO: 7. In other embodiments, the antibodies comprise a VH and a VL, wherein the VH comprises: a HCDR1 comprising the amino acid sequence of SEQ ID NO: 62 and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and the VL comprises: a LCDR1 comprising the amino acid sequence of SEQ ID NO: 7.

[0256] In some embodiments, the antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 221 or 225 and the amino acid sequence of SEQ ID NO: 222; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 223. Each of these paratope regions has been identified as contacting PAD4, as described in Examples and Figures 8A to 8BIn some embodiments, the antibody further comprises: a light chain constant region comprising the amino acid sequence of SEQ ID NO: 224.

[0257] In some embodiments, the antibody comprises a VH comprising a glycine at Kabat position 94 (Gly94). (See Figure 1E ). Thus, for example, the initially identified mouse anti-human 13 antibody contained a glycine at Kabat position 94, but several human framework regions do not contain a glycine. Therefore, in certain humanized antibodies, a back mutation is required to obtain a glycine at Kabat position 94. Gly94 is an amino acid residue in a loop adjacent to the VH CDR3 (i.e., the VH CDR3 loop). In some embodiments, Gly94 is associated with the flexibility and / or geometry of the VH CDR3 loop. In some embodiments, Gly94 can interact with the VH CDR3. In some embodiments, Gly94 is associated with the activity of anti-PAD4 antibodies. In some embodiments, mutation of Gly94 to a different amino acid (such as threonine) results in reduced flexibility of the VH CDR3 loop, an altered geometry of the VH CDR3 loop, reduced interaction between the amino acid at position 94 and the VH CDR3 loop, reduced binding of the anti-PAD4 antibody to PAD4, reduced activity of the anti-PAD4 antibody, increased binding of the anti-PAD4 antibody to extracellular matrix (ECM) proteins, an altered secondary structure of the anti-PAD4 antibody, reduced stability of the anti-PAD4 antibody, or any combination thereof. Thus, in some embodiments, the VH of the antibody comprises a glycine at Kabat position 94 (position 98 of SEQ ID NO: 10). (See Figure 1E , which depicts this residue position just before HCDR3).

[0258] In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70. In some embodiments, the antibody comprises both a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68, and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NO: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68, modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of any one of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70, modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.In some such cases: the antibody VH and VL further comprise: (a) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4 or 62, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, and a LCDR1 comprising the amino acid sequence of SEQ ID NO: 7; (b) a HCDR1 comprising the amino acid sequence of SEQ ID NO: 4 or 62, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 7, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, a LCDR3 comprising the amino acid sequence of SEQ ID NO: 9; or (c) the amino acid sequence of SEQ ID NO: 221 or 225 in VH and the amino acid sequence of SEQ ID NO: 222 in VL, further optionally, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 224. Thus, in such cases, the variations in VH and VL compared to the sequence identification numbers listed above are located in regions outside of these specific CDR or paratope sequences. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70.

[0259] In yet other embodiments, the antibody comprises both a VH comprising the amino acid sequence of any one of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68, and a VL comprising the amino acid sequence of any one of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70. Thus, by way of example, the following exemplary antibodies are within the scope of the invention:

[0260] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 10; and a VL comprising the amino acid sequence of SEQ ID NO: 12;

[0261] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 14; and a VL comprising the amino acid sequence of SEQ ID NO: 16;

[0262] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 18; and a VL comprising the amino acid sequence of SEQ ID NO: 20;

[0263] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 22; and a VL comprising the amino acid sequence of SEQ ID NO: 24;

[0264] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 26; and a VL comprising the amino acid sequence of SEQ ID NO: 28;

[0265] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 30; and a VL comprising the amino acid sequence of SEQ ID NO: 32;

[0266] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 34; and a VL comprising the amino acid sequence of SEQ ID NO: 36;

[0267] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 38; and a VL comprising the amino acid sequence of SEQ ID NO: 40;

[0268] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 42; and a VL comprising the amino acid sequence of SEQ ID NO: 44;

[0269] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 46; and a VL comprising the amino acid sequence of SEQ ID NO: 48;

[0270] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 50; and a VL comprising the amino acid sequence of SEQ ID NO: 52;

[0271] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 54; and a VL comprising the amino acid sequence of SEQ ID NO: 56;

[0272] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 58; and a VL comprising the amino acid sequence of SEQ ID NO: 60;

[0273] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 68; and a VL comprising the amino acid sequence of SEQ ID NO: 70.

[0274] In any of the above antibodies, in some cases, the antibody binds to an epitope on PAD4 comprising SEQ ID NO: 217 and SEQ ID NO: 218.

[0275] In some embodiments herein, the antibody is an IgA, IgG or IgM antibody. In some cases, the antibody is an IgG antibody, such as a human IgG1, IgG2, IgG3 or IgG4 antibody, or a murine IgG1 or IgG2 antibody. In some cases, the antibody comprises a wild-type, human IgG1, IgG2 or IgG4 heavy chain constant region. In some embodiments, the antibody comprises a full-length heavy chain and / or a full-length light chain. In other cases, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In other cases, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some cases, the antibody is an antibody fragment, such as Fv, single-chain Fv (scFv), Fab, Fab' or (Fab')2.

[0276] In some embodiments, the antibody is a bispecific or multispecific antibody, or it is covalently or non-covalently bound to at least one other molecule. In some embodiments, the antibody is covalently or non-covalently bound to at least one other molecule, wherein the at least one other molecule comprises a detection label and / or a drug.

[0277] In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some embodiments, the antibody comprises a light chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises both: a human IgG1 heavy chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194. In some instances, the antibody comprises a heavy chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some cases, the antibody comprises a light chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194. In some cases, the antibody comprises both a heavy chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192, and a light chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some embodiments, the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.In some embodiments, the antibody comprises both: a human IgG1 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.

[0278] Clone 20 and related antibodies

[0279] The present invention further relates to a second mouse anti-human antibody, clone 20, and variants thereof, hz20-1 to hz20-14, as described in the Examples below. Thus, in some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 73, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 74; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 75, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 76, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 77.

[0280] In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134, modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions, and wherein the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NOs: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136, modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some such cases, the antibody VH further comprises: a HCDR1 comprising the amino acid sequence of SEQ ID NO: 72, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 73, or a HCDR3 comprising the amino acid sequence of SEQ ID NO: 74. In some cases, the antibody VL further comprises: a LCDR1 comprising the amino acid sequence of SEQ ID NO: 75, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 76, or a LCDR3 comprising the amino acid sequence of SEQ ID NO: 77. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136.

[0281] In yet other embodiments, the antibody comprises a VH comprising the amino acid sequence of any one of SEQ ID NOs: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134, and a VL comprising the amino acid sequence of any one of SEQ ID NOs: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136. Thus, by way of example, the following exemplary antibodies are within the scope of the invention:

[0282] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 78; and a VL comprising the amino acid sequence of SEQ ID NO: 80;

[0283] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 82; and a VL comprising the amino acid sequence of SEQ ID NO: 84;

[0284] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 86; and a VL comprising the amino acid sequence of SEQ ID NO: 88;

[0285] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 90; and a VL comprising the amino acid sequence of SEQ ID NO: 92;

[0286] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 94; and a VL comprising the amino acid sequence of SEQ ID NO: 96;

[0287] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 98; and a VL comprising the amino acid sequence of SEQ ID NO: 100;

[0288] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 102; and a VL comprising the amino acid sequence of SEQ ID NO: 104;

[0289] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 106; and a VL comprising the amino acid sequence of SEQ ID NO: 108;

[0290] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 110; and a VL comprising the amino acid sequence of SEQ ID NO: 112;

[0291] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 114; and a VL comprising the amino acid sequence of SEQ ID NO: 116;

[0292] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 118; and a VL comprising the amino acid sequence of SEQ ID NO: 120;

[0293] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 122; and a VL comprising the amino acid sequence of SEQ ID NO: 124;

[0294] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 126; and a VL comprising the amino acid sequence of SEQ ID NO: 128;

[0295] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 130; and a VL comprising the amino acid sequence of SEQ ID NO: 132;

[0296] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 134; and a VL comprising the amino acid sequence of SEQ ID NO: 136.

[0297] In any of the above antibodies, in some cases, the antibody binds to an epitope on PAD4 comprising SEQ ID NO:219 and SEQ ID NO:220.

[0298] In some embodiments herein, the antibody is an IgA, IgG or IgM antibody. In some cases, the antibody is an IgG antibody, such as human IgG1, IgG2, IgG3 or IgG4 antibody, or murine IgG1 or IgG2 antibody. In some cases, the antibody comprises a wild-type, human IgG1, IgG2 or IgG4 heavy chain constant region. In some aspects, the antibody belongs to the human IgG1 isotype. In some aspects, the antibody belongs to the human IgG1 isotype, which has P329G, L234A and L235A mutations (LALAPG; EU numbering) to reduce Fc region effector function. In other aspects, the antibody belongs to the human IgG2 isotype. In some aspects, the antibody belongs to the IgG4 isotype, wherein the hinge region has an S228P mutation (EU numbering) to improve the stability of the IgG4 antibody. In some aspects, the antibody (e.g., non-humanized antibody) may have a non-human IgG constant region, and may be, for example, a murine IgG2a antibody, such as a murine IgG2a LAPG antibody. In some embodiments, the antibody comprises a full-length heavy chain and / or a full-length light chain. In other cases, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In yet other cases, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some cases, the antibody is an antibody fragment, such as Fv, single-chain Fv (scFv), Fab, Fab' or (Fab')2.

[0299] In some embodiments, the antibody is a bispecific or multispecific antibody, or it is covalently or non-covalently bound to at least one other molecule. In some embodiments, the antibody is covalently or non-covalently bound to at least one other molecule, wherein the at least one other molecule comprises a detection label and / or a drug.

[0300] In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some embodiments, the antibody comprises a light chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises both: a human IgG1 heavy chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194. In some instances, the antibody comprises a heavy chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some cases, the antibody comprises a light chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194. In some cases, the antibody comprises both a heavy chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192, and a light chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some embodiments, the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.In some embodiments, the antibody comprises both: a human IgG1 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.

[0301] pH-dependent clone 13-related antibodies

[0302] In some embodiments, the present invention relates to one or more antibody variants of the parent antibody clone 13 or hz13-5, wherein specific residues are modified as described below in Example 14. In some, but not all cases, these modifications were shown to affect the pH dependence of PAD4 binding, as described in Example 14.

[0303] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35; a HCDR2 comprising the amino acid sequence of positions 50 to 66; and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 142, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, or SEQ ID NO: 168. In some of the above embodiments, the antibody further comprises a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170; a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170; and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In other embodiments of the above, the antibody further comprises a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172; a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172; and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172.

[0304] In some embodiments, the antibody comprises:

[0305] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 138, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 138, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 138; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0306] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 140, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 140, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 140; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0307] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 142, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 142, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 142; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0308] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 144, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 144, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 144; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0309] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 146, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 146; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0310] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 148, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 148, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 148; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0311] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 150, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 150, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 150; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0312] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 152, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 152, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 152; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0313] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 154, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 154, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 154; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0314] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 156, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 156, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 156; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0315] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 158, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 158, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 158; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0316] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 160, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 160, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 160; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0317] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 162, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 162, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 162; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0318] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 164, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 164; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170;

[0319] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 166, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 166, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 166; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; or

[0320] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 168, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 168, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 168; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170.

[0321] In some embodiments, the antibody comprises:

[0322] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 138, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 138, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 138; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0323] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 140, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 140, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 140; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0324] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 142, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 142, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 142; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0325] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 144, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 144, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 144; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0326] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 146, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 146; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0327] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 148, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 148, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 148; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0328] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 150, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 150, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 150; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0329] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 152, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 152, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 152; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0330] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 154, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 154, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 154; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0331] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 156, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 156, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 156; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0332] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 158, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 158, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 158; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0333] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 160, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 160, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 160; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0334] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 162, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 162, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 162; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0335] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 164, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 164; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172;

[0336] - VH comprising: a HCDR1 comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 166, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 166, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 166; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; or

[0337] - VH comprising: a HCDR1 comprising the amino acid sequence at positions 26 to 35 of SEQ ID NO: 168, a HCDR2 comprising the amino acid sequence at positions 50 to 66 of SEQ ID NO: 168, and a HCDR3 comprising the amino acid sequence at positions 99 to 108 of SEQ ID NO: 168; and a VL comprising: a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172.

[0338] In some of the above embodiments, the antibody comprises a VH comprising an amino acid sequence that is at least 90% identical, at least 95% identical, or at least 97% identical to the amino acid sequence of SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 142, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, or SEQ ID NO: 168. In some of the above embodiments, the antibody comprises a VL comprising an amino acid sequence that is at least 90% identical, at least 95% identical, or at least 97% identical to the amino acid sequence of SEQ ID NO: 170. In other embodiments, the antibody comprises a VL comprising an amino acid sequence at least 90% identical, at least 95% identical, or at least 97% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody further comprises the corresponding HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 as provided above.

[0339] In some of the above embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 142, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166, or SEQ ID NO: 168. In some of the above embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some of the above embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 142, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166 or SEQ ID NO: 168; and a VL comprising the amino acid sequence of SEQ ID NO: 170. In some of the above embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 142, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQ ID NO: 166 or SEQ ID NO: 168; and a VL comprising the amino acid sequence of SEQ ID NO: 172.

[0340] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 138, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 138, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 138; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 138; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 138 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 138; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 138 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0341] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 138; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138 and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0342] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 138; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 138; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0343] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 140, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 140, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 140; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 140. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 140; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 140 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 140 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 140; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 140 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 140 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0344] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 140; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 140; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 140; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0345] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 140. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 140; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 140; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 140 and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0346] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 142, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 142, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 142; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 142. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 142; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 142 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 142 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 142; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 142 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 142 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0347] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 142. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 142; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 142; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 142; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0348] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 142. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 142; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 142; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 142; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0349] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 144, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 144, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 144; and a light chain variable region (VL) comprising: a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 144. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 144; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 144 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 144 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 144; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 144 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 144 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0350] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 144. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 144; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 144; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 144; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0351] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 144. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 144; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 144; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 144; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0352] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 146, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 146; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 146. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 146; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 146 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 146 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 146; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 146 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 146 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0353] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 146. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 146; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 146; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 146 and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0354] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 146. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 146; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 146; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 146; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0355] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 148, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 148, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 148; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 148. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 148; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 148 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 148 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 148; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 148 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 148 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0356] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 148. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 148; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 148; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 148; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0357] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 148. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 148; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 148; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 148; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0358] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 150, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 150, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 150; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 150. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 150; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 150 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 150 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 150, and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 150 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 150 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0359] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 150. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 150; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 150; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 150 and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0360] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 150. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 150; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 150; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 150 and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0361] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 152, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 152, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 152; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 152. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 152; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 152 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 152 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 152; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 152 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 152 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0362] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 152. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 152; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 152; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 152; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0363] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 152. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 152; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 152; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 152; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0364] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 154, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 154, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 154; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 154. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 154; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 154 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 154 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 154; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 154 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 154 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0365] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 154. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 154; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 154; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 154; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0366] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 154. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 154; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 154; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 154; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0367] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 156, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 156, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 156; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 156. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 156; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 156 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 156 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 156; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 156 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 156 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0368] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 156. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 156; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 156; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 156; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0369] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 156. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 156; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 156; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 156; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0370] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 158, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 158, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 158; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 158. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 158; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 158 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 158 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 158; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 158 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 158 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0371] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 158. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 158; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 158; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 158; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0372] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 158. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 158; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 158; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 158; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0373] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 160, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 160, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 160; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 160. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 160; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 160 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 160 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 160; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 160 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 160 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0374] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 160. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 160; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 160; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 160 and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0375] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 160. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 160; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 160; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 160 and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0376] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 162, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 162, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 162; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 162. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 162; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 162 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 162 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 162; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 162 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 162 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0377] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 162. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 162; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 162; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 162 and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0378] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 162. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 162; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 162; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 162; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0379] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 164, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 164; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 164. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 164; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 164 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 164 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 164; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 164 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 164 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0380] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 164. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 164; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 164; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 164; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0381] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 164. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 164; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 164; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 164; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0382] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 166, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 166, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 166; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 166. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 166; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 166 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 166 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 166; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 166 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 166 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0383] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 166. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 166; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 166; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 166; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0384] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 166. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 166; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 166; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 166; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0385] In some embodiments, the present invention relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises a heavy chain variable region (VH) comprising: a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26 to 35 of SEQ ID NO: 168, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 168, and a HCDR3 comprising the amino acid sequence of positions 99 to 108 of SEQ ID NO: 168; and a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 168. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 168; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 168 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 168 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172.In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 168; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 168 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises: a VL comprising the amino acid sequence of SEQ ID NO: 172 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acid sequence of SEQ ID NO: 168 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of SEQ ID NO: 172 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0386] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 168. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 168; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 168; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 168 and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0387] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 168. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 168; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 168; and a VL that comprises the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 168; and a VL that comprises the amino acid sequence of SEQ ID NO: 172.

[0388] In some cases, the antibody is an IgG antibody, such as a human IgG1, IgG2, IgG3 or IgG4 antibody, or a murine IgG1 or IgG2 antibody. In some cases, the antibody comprises a wild-type, human IgG1, IgG2 or IgG4 heavy chain constant region. In some embodiments, the antibody comprises a full-length heavy chain and / or a full-length light chain. In other cases, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In other cases, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some cases, the antibody is an antibody fragment, such as Fv, single-chain Fv (scFv), Fab, Fab' or (Fab')2.

[0389] In some embodiments, the antibody is a bispecific or multispecific antibody, or it is covalently or non-covalently bound to at least one other molecule. In some embodiments, the antibody is covalently or non-covalently bound to at least one other molecule, wherein the at least one other molecule comprises a detection label and / or a drug.

[0390] In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some embodiments, the antibody comprises a light chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises both: a human IgG1 heavy chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising an amino acid sequence at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 194. In some instances, the antibody comprises a heavy chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some cases, the antibody comprises a light chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194. In some cases, the antibody comprises both a heavy chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192, and a light chain constant region comprising an amino acid sequence modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some embodiments, the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.In some embodiments, the antibody comprises both: a human IgG1 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.

[0391] Mouse antibodies

[0392] In some embodiments, the present invention relates to an isolated antibody that specifically binds to murine protein arginine deiminase 4 (PAD4), wherein the antibody comprises a VH comprising: a HCDR1 comprising the amino acid sequence of positions 31 to 35 of SEQ ID NO: 208, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 208, and a HCDR3 comprising the amino acid sequence of positions 99 to 107 of SEQ ID NO: 208; and wherein the antibody comprises a light chain variable region (VL) comprising: a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 210, a LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 210, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 210.

[0393] In some embodiments, the antibody comprises a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 208. In some embodiments, the antibody comprises a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 210. In some embodiments, the antibody comprises both: a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 208; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 210. In some embodiments, the antibody comprises a VH comprising amino acids of SEQ ID NO: 208 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of any one of SEQ ID NO: 210 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some embodiments, the antibody comprises both: a VH comprising the amino acids of SEQ ID NO: 208 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions; and a VL comprising the amino acid sequence of any one of SEQ ID NO: 210 modified with 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some such cases, the antibody further comprises: a HCDR1 comprising the amino acid sequence of positions 31 to 35 of SEQ ID NO: 208, a HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 208, and a HCDR3 comprising the amino acid sequence of positions 99 to 107 of SEQ ID NO: 208; a LCDR1 comprising the amino acid sequence of residues 24 to 38 of SEQ ID NO: 210, a HCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 210, and a LCDR3 comprising the amino acid sequence of residues 93 to 101 of SEQ ID NO: 210.

[0394] In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 208. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 210. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 208 and a VL comprising the amino acid sequence of SEQ ID NO: 210.

[0395] In some embodiments herein, the antibody is an IgA, IgG or IgM antibody. In some cases, the antibody is a murine IgG1 or IgG2 antibody. In some embodiments, the antibody comprises a full-length heavy chain and / or a full-length light chain. In other cases, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In other cases, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some cases, the antibody is an antibody fragment, such as Fv, single-chain Fv (scFv), Fab, Fab' or (Fab')2.

[0396] In some embodiments, the antibody is a bispecific or multispecific antibody, or it is covalently or non-covalently bound to at least one other molecule. In some embodiments, the antibody is covalently or non-covalently bound to at least one other molecule, wherein the at least one other molecule comprises a detection label and / or a drug.

[0397] In some embodiments, the antibody comprises a HC comprising the amino acid sequence of SEQ ID NO: 212. In some embodiments, the antibody comprises a LC comprising the amino acid sequence of SEQ ID NO: 214. In some embodiments, the antibody comprises both: a HC comprising the amino acid sequence of SEQ ID NO: 212; and a LC comprising the amino acid sequence of SEQ ID NO: 214.

[0398] III. Exemplary Antibody Variants, Fragments, and Constant Regions

[0399] In many embodiments, antibodies that specifically bind to PAD4 can further incorporate any of the features, alone or in combination, as described in the following sections.

[0400] A. Antibody fragments

[0401] In certain embodiments, the antibodies provided herein are antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments, and other fragments described below. For a review of certain antibody fragments, see Hudson et al., Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, for example, Pluckthün, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds. (Springer-Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Pat. Nos. 5,571,894 and 5,'87,458. For a discussion of Fab and F(ab')2 fragments comprising salvage receptor binding epitope residues and having increased in vivo half-life, see U.S. Pat. No. 5,869,046.

[0402] Diabodies are antibody fragments in which both antigen-binding sites can be bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0403] Single-domain antibodies are antibody fragments that contain all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, the single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516).

[0404] Antibody fragments can be made by a variety of techniques as described herein, including, but not limited to, proteolytic digestion of intact antibodies and production by recombinant host cells (eg, E. coli or phage).

[0405] B. Bispecific or Multispecific Antibodies

[0406] In certain embodiments, the antibodies provided herein are multispecific antibodies, such as bispecific antibodies. Multispecific antibodies are monoclonal antibodies that have binding specificity for at least two different sites. In certain embodiments, one of the binding specificities is for TREM2 and the other is for any other antigen. In certain embodiments, bispecific antibodies can bind to two different epitopes of TREM2. Bispecific antibodies can also be used to locate drugs such as cytotoxic agents or to locate detection markers to cells expressing TREM2. In some embodiments, multispecific antibodies (e.g., bispecific antibodies) comprise a first variable domain containing a CDR or variable region as described herein. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[0407] Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see Milstein and Cuello, Nature 305:537 (1983)), WO 93 / 08829, and Traunecker et al., EMBO J. 10:3655 (1991)), and "knob-in-hole" engineering (see, e.g., U.S. Pat. No. 5,731,168). Multispecific antibodies can also be made by engineering electrostatic director effects for making antibody Fc-heterodimeric molecules (WO 93 / 08829). 2009 / 089004A1); cross-linking two or more antibodies or fragments (see, e.g., U.S. Patent No. 4,676,980 and Brennan et al., Science, 229:81 (1985)); using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 18(5):1547-1553 (1992)); using "bifunctional" antibodies for making bispecific antibody fragments. trispecific antibodies are prepared using "antibody" technology (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and as described, e.g., in Tutt et al., J. Immunol. 147:60 (1991).

[0408] Also included herein are engineered antibodies having three or more functional antigen binding sites, including "octopus antibodies" (see, e.g., US 2006 / 0025576).

[0409] C. Chimeric and humanized antibodies

[0410] In certain embodiments, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described in, for example, U.S. Patent No. 4,816,567; and in Morrison et al., Proc. Natl. Acad. Sci. USA (81: 6851-6855 (1984)). In one embodiment, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate (such as a monkey)) and a human constant region. In another embodiment, a chimeric antibody is a "class-switched" antibody, in which the class or subclass has changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0411] In certain embodiments, chimeric antibodies are humanized antibodies. Typically, non-human antibodies are humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parent non-human antibody. Generally speaking, a humanized antibody comprises one or more variable domains, wherein HVR, such as CDR (or a portion thereof), is derived from a non-human antibody, and FR (or a portion thereof) is derived from a human antibody sequence. The humanized antibody optionally also comprises at least a portion of a human constant region. In some embodiments, some FR residues in the humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., an antibody from which the HVR residues are derived) to, for example, restore or improve antibody specificity or affinity.

[0412] Humanized antibodies and methods for making them are reviewed, for example, in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described, for example, in Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991) (describing “resurfacing”); Dall'Acqua et al., Methods 36:43-60 (2005) (describing “FR shuffling”); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the “guided selection” method of FR shuffling).

[0413] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best fit" method (see, e.g., Sims et al. J. Immunol. 151: 2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA 2003; 1996; 1997; 2003; 2004; 2005; 2006; 2007; 2008; 2009; 2010; 2011; 2012; 2013; 2014; 2015; 2016; 2017; 2018; 2019; 2020; 2021; 2022; 2023; 2024; 2025; 2026; 2027; 2028; 2029; 2030; 2031; 2032; 2033; 2034; 2035; 2036; 2037; 2038; 2039; 2040; 2041; 2042; 2043; 2044; 2045; 2046; 2047; 2048; 2049; 2150; 2161; 2170; 2171; 2172; 2180 c. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived from screening FR repertoires (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0414] In some embodiments, a humanized antibody may comprise a human IgG1, IgG2, IgG3, or IgG4 heavy chain constant region.

[0415] D. Glycosylation and PEGylation variants

[0416] In certain embodiments, the glycosylation of the antibody is modified. For example, aglycosylated antibodies (i.e., antibodies lacking glycosylation) can be produced. Glycosylation can be altered to, for example, increase the affinity of the antibody for the antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made to eliminate one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation can increase the affinity of the antibody for the antigen. Such methods are further described in detail in U.S. Patent Nos. 5,714,350 and 6,350,861 to Co et al.

[0417] Glycosylation at N297 can be reduced by mutating the N297 residue to another residue (eg, N297A), and / or by mutating an adjacent amino acid (eg, 298) to prevent glycosylation of the constant region at N297.

[0418] Additionally or alternatively, antibodies with altered glycosylation patterns can be produced, such as hypofucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GlcNac structures. Such altered glycosylation patterns have been shown to increase the ADCC ability of antibodies. Such carbohydrate modifications can be achieved, for example, by expressing the antibody in a host cell with an altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells for expressing the recombinant antibodies described herein to thereby produce antibodies with altered glycosylation. For example, EP 1,176,195 by Hanai et al. describes a cell line in which the FUT8 gene encoding the fucosyltransferase is functionally disrupted, such that antibodies expressed in this cell line exhibit hypofucosylation. Presta's PCT publication WO 03 / 035835 describes a mutant CHO cell line, Led 3 cells, which has a reduced ability to attach fucose to carbohydrates attached to Asn(297), resulting in hypofucosylation of antibodies expressed in the host cells (see also Shields, RL et al., 2002 J. Biol. Chem. 277:26733-26740). Umana et al.'s PCT publication WO 99 / 54342 describes a cell line engineered to express a glycoprotein-modifying glycosyltransferase {e.g., β(1,4)-N-acetylglucosaminyltransferase III (GnTIII)), such that antibodies expressed in the engineered cell line exhibit increased bisecting GlcNac structures, resulting in increased ADCC activity of the antibodies (see also Umana et al. (1999) Nat. Biotech. 17:176-180).

[0419] Another modification of the antibodies described herein is pegylation. Antibodies can be pegylated to, for example, extend the biological (e.g., serum) half-life of the antibody. To pegylate an antibody, the antibody or its fragment is typically reacted with polyethylene glycol (PEG) (such as a reactive ester or aldehyde derivative of PEG) under conditions where one or more PEG groups are attached to the antibody or antibody fragment. In some embodiments, pegylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or a similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any PEG form that has been used to derive other proteins, such as single (CI-CIO) alkoxy or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is a non-glycosylated antibody. Protein pegylation methods are known in the art and can be applied to the antibodies described herein. See, for example, EP 0 154316 by Nishimura et al. and EP 0 401 384 by Ishikawa et al.

[0420] E. Constant region

[0421] In some embodiments, the antibodies described herein comprise one or more human constant regions. In some embodiments, the human heavy chain constant region belongs to an isotype selected from IgA, IgG, and IgD. In some embodiments, the human light chain constant region belongs to an isotype selected from κ and λ. In some embodiments, the antibodies described herein comprise a human IgG constant region, such as IgG1, IgG2, IgG3, or IgG4. In some embodiments, the antibodies described herein comprise a human IgG4 heavy chain constant region. In some such embodiments, the antibodies described herein comprise an S241P mutation in a human IgG4 constant region. In some embodiments, the antibodies described herein comprise a human IgG4 constant region and a human κ light chain.

[0422] The choice of heavy chain constant region can determine whether the antibody will have effector functions in vivo. In some embodiments, such effector functions include antibody-dependent cell-mediated cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), and can result in killing cells to which the antibody is bound. In some therapeutic methods, including methods for treating some cancers, for example, when an antibody binds to cells that support tumor maintenance or growth, it may be necessary to kill the cells. Exemplary cells that can support tumor maintenance or growth include, but are not limited to, tumor cells themselves, cells that help recruit blood vessels to tumors, and cells that provide ligands, growth factors, or counter-receptors that support or promote tumor growth or tumor survival. In some embodiments, when effector functions are needed, antibodies comprising human IgG1 heavy chains or human IgG3 heavy chains are selected.

[0423] In certain embodiments, the antibodies provided herein are altered to increase or decrease the degree of antibody glycosylation. Adding glycosylation sites to an antibody or deleting glycosylation sites from an antibody can be conveniently achieved by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0424] In the case where the antibody comprises an Fc region, the carbohydrates attached thereto may be changed. Natural antibodies produced by mammalian cells typically comprise branched biantennary oligosaccharides, which are typically linked to Asn297 of the CH2 domain of the Fc region via an N-bond. See, for example, Wright et al., TIBTECH 15:26-32 (1997). Oligosaccharides may include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose linked to the GlcNAc in the "backbone" of the biantennary oligosaccharide structure. In some embodiments, the oligosaccharides in the antibodies of the present invention may be modified to form antibodies with certain improved properties. For example, in some embodiments, antibodies may be defucosylated, for example, by mutating residues such as Asn297 that are typically glycosylated with fucose or glycosylated by other means. In some embodiments, the antibodies herein may comprise a defucosylated human IgG1 constant region.

[0425] The antibodies further have bisected oligosaccharides, for example, bi-tactile oligosaccharides linked to the Fc region of the antibody bisected by GlcNAc. Such antibodies may have reduced fucosylation and / or improved ADCC function. Examples of such antibodies are described, for example, in WO 2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and US 2005 / 0123546 (Umana et al.). Antibodies in which at least one galactose residue in the oligosaccharide is linked to the Fc region are also provided. Such antibodies may have improved CDC function. Such antibodies are described, for example, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).

[0426] Antibodies with amino-terminal leader sequence extensions are also provided. For example, one or more amino acid residues of the amino-terminal leader sequence are present at the amino terminus of any one or more heavy or light chains of the antibody. Exemplary amino-terminal leader sequence extensions comprise or consist of three amino acid residues VHS, which are present on one or both light chains of the antibody.

[0427] The in vivo or serum half-life of human FcRn high affinity binding polypeptides can be analyzed, for example, in transgenic mice, humans, or non-human primates administered polypeptides with variant Fc regions. See also, for example, Petkova et al. International Immunology 18(12):1759-1769 (2006).

[0428] In some embodiments of the invention, the defucosylated antibody mediates ADCC more efficiently in the presence of human effector cells compared to the parent antibody comprising fucose. Generally, ADCC activity can be determined using an in vitro ADCC assay as disclosed herein, but other assays or methods for determining ADCC activity, for example, in animal models, are contemplated.

[0429] In certain embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to change the effector function of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 297, 318, 320, 322, 330 and / or 331 (EU numbering) may be replaced with different amino acid residues to change the affinity of the antibody for the effector ligand, but retain the antigen binding ability of the parent antibody. The effector ligand with altered affinity may be, for example, an Fc receptor or the C1 component of complement. This method is further described in detail in U.S. Patent Nos. 5,624,821 and 5,648,260 to Winter et al.

[0430] In some embodiments, one or more amino acids selected from amino acid residues 329, 331, and 322 can be replaced with a different amino acid residue such that the antibody has altered C1q binding and / or reduced or abolished complement-dependent cytotoxicity (CDC). This approach is further described in U.S. Patent No. 6,194,551 to Idusogie et al.

[0431] In some embodiments, one or more amino acid residues within amino acid positions 231 and 239 are altered to alter the ability of the antibody to fix complement. This approach is further described, for example, in PCT Publication WO 94 / 29351 by Bodmer et al. In some embodiments, the Fc region can be modified to reduce antibody-dependent cellular cytotoxicity (ADCC) and / or reduce affinity for Fcγ receptors by modifying one or more amino acids at the following positions: 234, 235, 236, 238, 239, 240, 241, 243, 244, 245, 247, 248, 249, 252, 254, 255, 256, 258, 262, 263, 264, 265, 267, 268, 269, 270, 272, 276, 278, 280, 283, 285, 286, 289, 290 , 292, 293, 294, 295, 296, 298, 299, 301, 303, 305, 307, 309, 312, 313, 315, 320, 322, 324, 325, 326, 327, 329, 330, 331, 332, 333, 334, 335, 337, 338, 340, 360, 373, 376, 378, 382, ​​388, 389, 398, 414, 416, 419, 430, 433, 434, 435, 436, 437, 438 or 439 (EU number). Exemplary substitutions include 236A, 239D, 239E, 268D, 267E, 268E, 268F, 324T, 332D, and 332E. Exemplary variants include 239D / 332E, 236A / 332E, 236A / 239D / 332E, 268F / 324T, 267E / 268F, 267E / 324T, and 267E / 268F / 324T (EU numbering). Other Fc modifications that can be made to Fcs are those that reduce or eliminate binding to FcγRs and / or complement proteins, thereby reducing or eliminating Fc-mediated effector functions such as ADCC, ADCP, and CDC. Exemplary modifications include, but are not limited to, substitutions, insertions, and deletions at positions 234, 235, 236, 237, 267, 269, 325, 328, 330, and / or 331 (e.g., 330 and 331), where numbering is according to the EU index. Exemplary substitutions include, but are not limited to, 234A, 235E, 236R, 237A, 267R, 269R, 325L, 328R, 330S, and 331S (e.g., 330S and 331S), where numbering is according to the EU index. The Fc variant may comprise 236R / 328R.Other modifications used to reduce the interaction of FcγRs with complement include substitutions 297A, 234A, 235A, 237A, 318A, 228P, 236E, 268Q, 309L, 330S, 331S, 220S, 226S, 229S, 238S, 233P, and 234V, as well as removal of glycosylation at position 297 by mutation or enzymatic means or by production in an organism that does not glycosylate proteins, such as bacteria. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20: 685-691. For example, the human IgG1.3 Fc constant region contains the substitutions L234A, L235E, and G237A. IgG1fa.P238K (or IgG1.P238K) contains the substitution P238K. IgG1.1f contains the substitutions L234A, L235E, G237A, A330S, and P331S (all numbering is according to the EU index).

[0432] Fc variants that enhance affinity for the inhibitory receptor FcγRIIb can also be used. Such variants can provide Fc fusion proteins with immunomodulatory activity associated with FcγRIIb cells, including, for example, B cells and monocytes. In one embodiment, the Fc variant provides a selectively enhanced affinity for FcγRIIb relative to one or more activating receptors. Modifications for altering binding to FcγRIIb include one or more modifications at positions selected from the group consisting of: 234, 235, 236, 237, 239, 266, 267, 268, 325, 326, 327, 328, 330, 331, and 332 according to the EU index. Exemplary substitutions for enhancing affinity for FcγRllb include, but are not limited to, 234A, 234D, 234E, 234F, 234W, 235D, 235E, 235F, 235R, 235Y, 236D, 236N, 237A, 237D, 237N, 239D, 239E, 266M, 267D, 267E, 268D, 268E, 327D, 327E, 328F, 328W, 328Y, 330S, 331S, and 332E. Exemplary substitutions include 235Y, 236D, 239D, 266M, 267E, 268D, 268E, 328F, 328W, and 328Y. Other Fc variants for enhanced binding to FcγRIIb include 235Y / 267E, 236D / 267E, 239D / 268D, 239D / 267E, 267E / 268D, 267E / 268E, and 267E / 328F. (All numbering is under the EU index).

[0433] Other modifications used to enhance the interaction of FcγRs with complement include, but are not limited to, substitutions 298A, 333A, 334A, 326A, 2471, 339D, 339Q, 280H, 290S, 298D, 298V, 243L, 292P, 300L, 396L, 3051, and 396L. These and other modifications are reviewed in Strohl, 2009, Current Opinion in Biotechnology 20: 685-691. Fc modifications that increase binding to Fcγ receptors include amino acid modifications at any one or more of the following amino acid positions in the Fc region: 238, 239, 248, 249, 252, 254, 255, 256, 258, 265, 267, 268, 269, 270, 272, 279, 280, 283, 285, 298, 289, 290, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325 6, 298, 301, 303, 305, 307, 312, 315, 324, 327, 329, 330, 335, 337, 338, 340, 360, 373, 376, 379, 382, ​​388, 389, 398, 414, 416, 419, 430, 434, 435, 437, 438 or 439, wherein the numbering of the Fc region residues is that of the EU index as in patent publication No. WO 00 / 42072.

[0434] Optionally, the Fc region may comprise non-naturally occurring amino acid residues at additional and / or alternative positions known to those skilled in the art (see, e.g., U.S. Pat. Nos. 5,624,821, 6,277,375, 6,737,056, 6,194,551, 7,317,091, 8,101,720; PCX patent publications WO 00 / 42072, WO 01 / 58957, WO 02 / 06919, WO 04 / 016750, WO 04 / 029207, WO 04 / 035752, WO 04 / 074455, WO 04 / 099249, WO 04 / 063351, WO 05 / 070963, WO 05 / 040217, WO 05 / 092925, and WO 06 / 020114).

[0435] The affinity and binding properties of the Fc region for its ligand can be determined by various in vitro analytical methods known in the art (biochemical or immunological based assays), including but not limited to equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)) or kinetics (e.g., BIACORE assays), as well as other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). These and other methods may utilize labels on one or more of the components being assayed and / or employ a variety of detection methods, including but not limited to chromogenic, fluorescent, luminescent, or isotopic labels. A detailed description of binding affinity and kinetics can be found in Paul, WE, ed., Fundamental Immunology, 4th edition, Lippincott-Raven, Philadelphia (1999), which focuses on antibody-immunogen interactions.

[0436] In certain embodiments, antibodies are modified to extend their biological half-life. Various methods are possible. For example, this can be achieved by increasing the binding affinity of the Fc region for FcRn. For example, one or more of the following residues can be mutated: 252, 254, 256, 433, 435, 436, as described in U.S. Patent No. 6,277,375. Specific exemplary substitutions include one or more of the following: T252L, T254S, and / or T256F. Alternatively, to extend biological half-life, antibodies can be modified within the CH1 or CL regions to contain salvage receptor binding epitopes derived from two loops of the CH2 domain of the Fc region of IgG, as described in U.S. Patent Nos. 5,869,046 and 6,121,022 to Presta et al. Other exemplary variants that increase binding to FcRn and / or improve pharmacokinetic properties include substitutions at positions 259, 308, 428, and 434, including, for example, 259I, 308F, 428L, 428M, 434S, 434I, 434F, 434Y, and 434X1. Other variants that increase Fc binding to FcRn include: 250E, 250Q, 428L, 428F, 250Q / 428L (Hinton et al. 2004, J. Biol. Chem. 279(8):6213-6216, Hinton et al. 2006 Journal of Immunology 176:346-356), 256A, 272A, 286A, 305A, 307A, 307Q, 311A, 312A, 376A, 378Q, 380A, 382A, 434A (Shields et al., Journal of Biological Chemistry 2006, 11:117-119). Chemistry,2001,276(9):6591-6604), 252F, 252T, 252Y, 252W, 254T, 256S, 256R, 256Q, 256E, 256D, 256T, 309P, 31 1S, 433R, 433S, 4331, 433P, 433Q, 434H, 434F, 434Y, 252Y / 254T / 256E, 433K / 434F / 436H, 308T / 309P / 311S (Journal of Dall'Acqua et al. Immunology, 2002, 169: 5171-5180, Dal'Acqua et al., 2006, Journal of Biological Chemistry Other modifications for modulating FcRn binding are described in Yeung et al., 2010, J Immunol, 182:7663-7671.

[0437] In certain embodiments, hybrid IgG isotypes with specific biological characteristics can be used. For example, IgG1 / IgG3 hybrid variants can be constructed by replacing the IgG1 position in CH2 and / or CH3 district with amino acids from IgG3 at different positions of two isotypes. Therefore, hybrid variant IgG antibodies comprising one or more substitutions, such as 274Q, 276K, 300F, 339T, 356E, 358M, 384S, 392N, 397M, 4221, 435R and 436F can be constructed. In some embodiments described herein, IgG1 / IgG2 hybrid variants can be constructed by replacing the IgG2 position in CH2 and / or CH3 district with amino acids from IgG1 at different positions of two isotypes. Therefore, hybrid variant IgG antibodies comprising one or more substitutions, such as one or more of the following amino acid substitutions, can be constructed: 233E, 234L, 235L, +236G (referring to the insertion of glycine at position 236) and 327A.

[0438] Furthermore, the binding sites for FcγRI, FcγRII, FcγRIII, and FcRn on human IgG1 have been mapped, and variants with improved binding have been described (see Shields, RL et al. (2001), J. Biol. Chem. 276:6591-6604). Specific mutations at positions 256, 290, 298, 333, 334, and 339 have been shown to improve binding to FcγRIII. Furthermore, the following combination mutants have been shown to improve FcγRIII binding: T256A / S298A, S298A / E333A, S298A / K224A, and S298A / E333A / K334A, and have been shown to exhibit enhanced FcγRIIIa binding and ADCC activity (Shields et al., 2001). Other IgG1 variants with strongly enhanced binding to FcγRIIIa have been identified, including variants with S239D / I332E and S239D / I332E / A330L mutations, which showed the greatest increase in affinity for FcγRIIIa, decreased binding to FcγRIIb, and potent cytotoxic activity in cynomolgus monkeys (Lazar et al., 2006). The introduction of the triple mutation into antibodies such as alemtuzumab (CD52 specific), trastuzumab (HER2 / neu specific), rituximab (CD20 specific), and cetuximab (EGFR specific) translated into greatly enhanced in vitro ADCC activity, and the S239D / I332E variant showed enhanced ability to deplete B cells in monkeys (Lazar et al., 2006). In addition, IgG1 mutants containing L235V, F243L, R292P, Y300L, and P396L mutations have been identified in transgenic mice expressing human FcγRIIIa in models of B-cell malignancies and breast cancer that exhibit enhanced binding to FcγRIIIa and concomitant enhanced ADCC activity (Stavenhagen et al., 2007; Nordstrom et al., 2011). Other Fc mutants that may be used include: S298A / E333A / L334A, S239D / I332E, S239D / I332E / A330L, L235V / F243L / R292P / Y300L / P396L, and M428L / N434S.

[0439] In certain embodiments, an Fc is selected that has reduced binding to FcγRs. An exemplary Fc with reduced FcγR binding, such as an IgG1 Fc, comprises the following three amino acid substitutions: L234A, L235E, and G237A.

[0440] In certain embodiments, an Fc is selected that has reduced complement binding. An exemplary Fc with reduced complement binding, for example, an IgG1 Fc has the following two amino acid substitutions: A330S and P331S.

[0441] In certain embodiments, an Fc is selected that has substantially no effector function, ie, it has reduced binding to FcγRs and reduced complement fixation. An exemplary non-effector Fc, such as an IgG1 Fc, comprises the following five mutations: L234A, L235E, G237A, A330S, and P331S.

[0442] When an IgG4 constant domain is used, it may include the substitution S228P, which mimics the hinge sequence in IgG1 and thereby stabilizes the IgG4 molecule.

[0443] Fc modifications described in WO 2017 / 087678 or WO2016081746 may also be used.

[0444] In certain embodiments, the glycosylation of the antibody is modified. For example, an aglycosylated antibody (i.e., an antibody lacking glycosylation) can be produced. Glycosylation can be altered to, for example, increase the affinity of the antibody for the antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made to eliminate one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site. Such aglycosylation can increase the affinity of the antibody for the antigen. This approach is further described in U.S. Patent Nos. 5,714,350 and 6,350,861 to Co et al.

[0445] Glycosylation at N297 can be reduced by mutating the N297 residue to another residue (eg, N297A), and / or by mutating an adjacent amino acid (eg, 298) to prevent glycosylation of the constant region at N297.

[0446] Additionally or alternatively, antibodies with altered glycosylation patterns can be produced, such as hypofucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GlcNac structures. Such altered glycosylation patterns have been shown to increase the ADCC ability of antibodies. Such carbohydrate modifications can be achieved, for example, by expressing the antibody in a host cell with an altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells for expressing the recombinant antibodies described herein to thereby produce antibodies with altered glycosylation. For example, EP 1,176,195 to Hanai et al. describes a cell line in which the FUT8 gene encoding the fucosyltransferase is functionally disrupted, such that antibodies expressed in this cell line exhibit hypofucosylation. Presta's PCT publication WO 03 / 035835 describes a mutant CHO cell line, Led 3 cells, which has a reduced ability to attach fucose to carbohydrates attached to Asn(297), resulting in hypofucosylation of antibodies expressed in the host cells (see also Shields, RL et al., 2002 J. Biol. Chem. 277:26733-26740). Umana et al.'s PCT publication WO 99 / 54342 describes a cell line engineered to express a glycoprotein-modifying glycosyltransferase {e.g., β(1,4)-N-acetylglucosaminyltransferase III (GnTIII)), such that antibodies expressed in the engineered cell line exhibit increased bisecting GlcNac structures, resulting in increased ADCC activity of the antibodies (see also Umana et al. (1999) Nat. Biotech. 17:176-180).

[0447] Another modification of the antibodies described herein is pegylation. Antibodies can be pegylated to, for example, extend the biological (e.g., serum) half-life of the antibody. To pegylate an antibody, the antibody or its fragment is typically reacted with polyethylene glycol (PEG) (such as a reactive ester or aldehyde derivative of PEG) under conditions where one or more PEG groups are attached to the antibody or antibody fragment. In some embodiments, pegylation is carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or a similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any PEG form that has been used to derive other proteins, such as single (CI-CIO) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is a non-glycosylated antibody. Protein pegylation methods are known in the art and can be applied to the antibodies described herein. See, for example, EP 0 154 316 by Nishimura et al. and EP 0 401 384 by Ishikawa et al.

[0448] IV. Exemplary Properties of Specific Anti-PAD4 Antibodies

[0449] In many embodiments, an antibody that specifically binds to PAD4 can comprise any of the following properties, alone or in combination.

[0450] In some cases, binding of an antibody to a ligand such as PAD4 can be determined by surface plasmon resonance (SPR). In some embodiments, the antibody binds to a ligand such as PAD4 with a K of less than 5 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.05 nM to 1 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.1 nM, or 0.5 nM to 1 nM in an SPR assay. D Specifically binds to PAD4. For example, this K D In some embodiments, the antibody has a K of less than 5 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.05 nM to 1 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.1 nM, or 0.5 nM to 1 nM in the presence of 1 to 2 mM calcium chloride (e.g., in the presence of 1 mM calcium chloride). D Specifically binds to PAD4. In some embodiments, the antibody is expressed by SPR in the presence of 1 to 2 mM calcium chloride (e.g., at 1 mM calcium chloride) and in the absence of calcium ions (due to the absence of additional calcium salts and the presence of EDTA, such as 1 mM or 2 mM EDTA) at a K of D Specifically binds to PAD4: less than 5 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.05 nM to 1 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.1 nM, or 0.5 nM to 1 nM. Thus, in this case, the antibody specifically binds to PAD4 both in the presence and absence of calcium ions (such as calcium chloride). For example, see Tables 2 and 3 herein with respect to the Examples.

[0451] In some embodiments, the anti-PAD4 antibody does not bind to human protein arginine deiminase 2 (PAD2). Thus, in a binding assay such as by SPR, binding to PAD2 is too weak to be detected. In some cases, the antibody specifically binds to human PAD4, for example, as just described above, but does not bind to murine PAD4 (i.e., no binding to murine PAD4 is detected in an SPR or similar assay). In some cases, the antibody specifically binds to human PAD4, for example, as just described above, but does not bind to macaque PAD4 (i.e., no binding to macaque PAD4 is detected in an SPR assay), while in other cases, the antibody specifically binds to human and macaque PAD4 in an SPR assay. In some cases, by SPR, the antibody specifically binds to human and cynomolgus macaque PAD4, but does not bind to murine PAD4. In other cases, the antibody specifically binds to human PAD4, but does not bind to macaque or murine PAD4, as determined by SPR.

[0452] In some embodiments, the ECM score of the antibody in the ECM analysis is less than 50, less than 30, less than 10, less than 5, 1 to 30, 1 to 20, 1 to 10, 1 to 5, 1 to 3, 1, 2, 3, 4 or 5. (See, e.g., Table 5). As described in Example 5 below, the ECM score indicates the extent to which the antibody nonspecifically binds to the extracellular matrix (ECM). Pre-coated ECM culture plates (such as commercially available 96-well plates) can be used for analysis. The ECM score can be determined by incubating the antibody in an ECM-coated plate for 1 hour, then incubating with an HRP-conjugated detection antibody and reacting with TMB substrate, and then dividing the absorbance value measured at 450 nm by the absorbance value of the control well to which the antibody was not added. (See Example 5 below).

[0453] In some embodiments, the antibodies herein have a hydrophobic interaction chromatography (HIC) retention time of 9 to 11 minutes on a TSKgel Butyl-NPR column (4.6 mm x 3.5 cm, 2.5 μm particle size, Tosoh P / N 14947) using a linear gradient of mobile phase A (0.1 M sodium phosphate, pH 7.0, 2 M ammonium sulfate) and mobile phase B (0.1 M sodium phosphate solution, pH 7.0) over 20 min at a flow rate of 1.0 ml / min at a column temperature of 25° C. (See Example 6 below). In some embodiments, the antibodies have a melting temperature (Tm) between 60 and 70° C. at both pH 6.0 and pH 8.3 as measured by intrinsic fluorescence. In some embodiments, the aggregation temperature (Tagg) of the antibody is between 64°C and 75°C at pH 6.0 and between 60°C and 70°C at pH 8.3 as measured by static light scattering (see Example 7).

[0454] In some embodiments, using Antibodies have relatively low immunogenicity as measured by immunogenicity testing. Commercial computer simulation immunogenicity risk assessment algorithm Peptide MHC class II binding was ranked across eight human HLA DRB1 allele supertypes to cover >90% of the variability present in the human population (De Groot and Martin, Clin Immunol, 2009, 131(2): 189-201). (See Example 15). For example, a drug (alemtuzumab), (rituximab), (daclizumab), (CDP-571), (gemtuzumab) and The in silico immunogenicity scores for (bevacizumab) ranged from 0% to 45%, as shown in Table 10 below, where (alemtuzumab), (rituximab), (daclizumab) and (CDP-571) are each in the range of 45% to 7%. In contrast, in some embodiments, the antibodies herein have an immunogenicity score in the same assay that is in the range of 0 to 5%, such as 0 to 2% or 0 to 1%. Certain antibodies herein have an immunogenicity score of 0%. (See Table 10). Thus, in some cases, the immunogenicity of the antibodies is lower than that of the antibodies as measured by computer-simulated immunogenicity assays. (alemtuzumab), (rituximab) and (daclizumab), one, two, or all three.

[0455] In some embodiments, the antibody inhibits the enzymatic activity of PAD4 in vitro. For example, in some cases, the antibody inhibits the conversion of arginine to citrulline in the peptide substrate TSTGGRQGSHH (SEQ ID NO: 216) by PAD4 in vitro with an IC50 of 10 to 200 nM, 50 to 200 nM, 10 to 100 nM, 20 to 100 nM, or 50 to 100 nM. (See Example 4, Table 4). For example, in some cases, the antibody inhibits the conversion of arginine to citrulline in the peptide substrate TSTGGRQGSHH (SEQ ID NO: 216) by PAD4 in vitro. For example, in some cases, inhibition is optionally performed in a dose-dependent manner with an IC50 of 0.1 to 10 nM (such as 0.2 to 5 nM), wherein the PAD4 concentration is 1 to 8 μg / mL. (See, e.g., Example 16 below).

[0456] In some embodiments, the antibody inhibits the enzymatic activity of PAD4 in vitro in the presence of endogenous anti-PAD4 antibodies (e.g., a polyclonal mixture of endogenous anti-PAD4 antibodies) from a human individual (e.g., a patient with a disease (e.g., a disease disclosed herein, e.g., an inflammatory or autoimmune disease, e.g., rheumatoid arthritis)). Endogenous anti-PAD4 antibodies may include or may be, for example, PAD4 inhibitory antibodies and / or PAD4 activating antibodies. In some embodiments, the antibody inhibits the enzymatic activity of PAD4 in vitro in the presence of endogenous antibodies that bind to and activate human PAD4 (activating antibodies). In some embodiments, the activating antibodies are antibodies derived from a patient with rheumatoid arthritis. In some embodiments, the activating antibodies are antibodies that cross-react with human PAD3 and human PAD4.

[0457] In some cases, the antibody reduces the amount of extracellular citrullinated histone H3 in LPS-stimulated human blood monocytes compared to an isotype control antibody. In some cases, the antibody reduces the secretion of GM-CSF in LPS-stimulated human blood monocytes compared to an isotype control antibody. In some cases, the antibody reduces the gene expression of GM-CSF in LPS-stimulated human blood monocytes compared to an isotype control antibody. In some cases, the antibody has two of these three properties. In some cases, the antibody has all three of these properties. In some embodiments, the LPS-stimulated human blood monocytes are CD14+CD16- monocytes isolated from fresh human PBMCs. (See Example 17 herein). In some embodiments, the antibody is capable of being internalized by LPS-stimulated human blood monocytes. For example, the antibody can be labeled with a dye to detect monocyte internalization. In some cases, the antibody can be internalized by LPS-stimulated CD14+ human monocytes. (See Example 18). These results demonstrate that the anti-PAD4 antibodies herein can function intracellularly within monocytes and, in some embodiments, can block PAD4 function both extracellularly and intracellularly.

[0458] In some embodiments, the antibody inhibits PAD4 from playing a role in inflamed lungs. For example, this can be demonstrated by a reduction in the citrullination of histone H3 or ITIH4 in bronchoalveolar lavage fluid (BALF) collected from the lungs, as described in Examples 20 or 25 herein. For example, BALF can be collected from mice with acute or chronic lung inflammation and analyzed for the amount of citrullinated H3 or ITIH4 protein in the presence of the antibody and in the presence of a wild-type control. In some embodiments, the mouse is a human PAD4 knock-in mouse. In some cases, compared to an isotype control (IC) antibody, in the presence of the antibody, the amount of citrullinated H3 in the human PAD4 knock-in mouse is reduced by at least 15%, at least 30%, at least 40%, at least 50% or at least 60%, and / or the amount of citrullinated ITIH4 is reduced by at least 50%, at least 70% or at least 80%, for example, as calculated by the following formula: Among them, the untreated The amount of citrullinated H3 or ITIH4 in mice, mice receiving IC, and mice receiving anti-PAD4 antibody.

[0459] In some embodiments, the antibody inhibits PAD4 from playing a role in inflamed joints, for example, as described in Examples 21, 22, and / or 26. This can be demonstrated, for example, by reducing the citrullination of ITIH4 and / or PRG4 in joint tissue in a mouse model of acute or chronic joint inflammation (such as inflammation induced by joint injection of LPS). In some cases, the mouse can be a human PAD4 knock-in mouse. In some cases, the antibody reduces the amount of extracellular citrullinated PRG4 and / or citrullinated ITIH4 in the mouse patella in an LPS-induced acute joint injury model in human PAD4 knock-in mice compared to an isotype control antibody. In some cases, the EC50 for reducing citrullinated ITIH4 and / or citrullinated PRG4 is 2 nM or less, 1 nM or less, 0.1 nM or less, 0.05 to 2 nM, or 0.1 to 1 nM.

[0460] In some embodiments, the antibodies herein can reduce pristane-induced extracellular trap formation of neutrophils (NETosis) and / or extracellular trap formation of monocytes (METosis). (See Example 23). METosis is the process of releasing extracellular traps composed of cellular DNA interspersed with histones and cellular proteins from monocytes or macrophages. This network of substances formed by METosis or NETosis (neutrophil origin) is important for defending against microorganisms and is also a major driver of autoimmune pathology and sterile inflammation. For example, pristane can be injected intraperitoneally into mice (such as human PAD4 gene knock-in mice) after treatment with the antibodies herein or with isotype control antibodies. In some cases, in such pristane-induced mouse models, the antibodies herein can reduce H3 citrullination in neutrophils, monocytes, M1 macrophages and / or M2 macrophages in the peritoneal fluid compared to the isotype control antibodies. In other cases, the antibodies can also reduce the amount of soluble markers of neutrophils and monocytes / macrophages in the peritoneal fluid of mice, such as elastase, MPO, MIP-2α, GROα / KC, MCP1, MIP1β, IL6, and MIP3α (see Example 23).

[0461] In some embodiments, the antibodies herein can inhibit PAD4-dependent responses in a collagen-induced arthritis mouse model. (See Example 24). For example, in some embodiments, the antibodies herein significantly reduce the clinical score of arthritis in mice in an arthritis model compared to an isotype control according to the following scale: (1) normal; (2) mild, with clear redness and swelling in the ankle or wrist, or limited to a single toe, regardless of the number of toes affected; (3) moderate redness and swelling in the ankle or wrist; (4) severe redness and swelling of the entire paw, including the toes; (5) the most severe inflammation in the limb, involving multiple joints. For example, in some cases, the antibodies herein also reduce NETosis and METosis as assessed by SG+MPO+neutrophils and SG+MPO+monocytes / macrophages, respectively, and / or reduce soluble markers of monocytes / macrophages, such as elastase and MPO, and / or reduce the proportion of H3 protein citr...

Claims

1. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:62 and a HCDR3 comprising the amino acid sequence of SEQ ID NO:6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:

7.

2. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:62, a HCDR2 comprising the amino acid sequence of SEQ ID NO:5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO:6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:7, a LCDR2 comprising the amino acid sequence of SEQ ID NO:8, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

9.

3. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 225 and the amino acid sequence of SEQ ID NO: 222; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:

223.

4. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:62 and a HCDR3 comprising the amino acid sequence of SEQ ID NO:6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:7, wherein the VH of the antibody is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO:

68.

5. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising a heavy chain complementary determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:62, a HCDR2 comprising the amino acid sequence of SEQ ID NO:5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO:6; and a light chain variable region (VL) comprising a light chain complementary determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:7, a LCDR2 comprising the amino acid sequence of SEQ ID NO:8, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:9, wherein the VH of the antibody is at least 90%, at least 95%, at least 97%, or at least 99% identical to the amino acid sequence of SEQ ID NO:

68.

6. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 225 and the amino acid sequence of SEQ ID NO: 222; and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 223, wherein the VH of the antibody is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO:

68.

7. The isolated antibody according to any one of claims 1 to 4, wherein the VL of the antibody is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO:

70.

8. The isolated antibody according to any one of claims 1 to 7, wherein the heavy chain variable region comprises a glycine at Kabat position 94, which position corresponds to position 98 of SEQ ID NO:

68.

9. The isolated antibody according to any one of claims 1 to 8, wherein the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO:

68.

10. The isolated antibody according to any one of claims 1 to 8, wherein the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:

70.

11. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 68; and a VL comprising the amino acid sequence of SEQ ID NO:

70.

12. The isolated antibody according to any one of claims 1 to 11, wherein the antibody binds to an epitope on PAD4 comprising SEQ ID NO: 217 and optionally SEQ ID NO:

218.

13. An isolated antibody according to any one of claims 1 to 12 which is an IgA, IgG or IgM antibody.

14. An isolated antibody according to any one of claims 1 to 12, which is an IgG antibody, such as a human IgG1, IgG2, IgG3 or IgG4 antibody.

15. The isolated antibody according to claim 14, wherein the antibody comprises a wild-type, human IgGl, IgG2 or IgG4 heavy chain constant region.

16. An isolated antibody according to any one of claims 1 to 12, wherein the antibody comprises a human IgG1 or IgG2 heavy chain constant region comprising an amino acid sequence that is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190 or 192.

17. An isolated antibody according to any one of claims 1 to 12 or 16, wherein the antibody comprises a light chain constant region comprising an amino acid sequence that is at least 90%, at least 95%, at least 97% or at least 99% identical to the amino acid sequence of SEQ ID NO:

194.

18. An isolated antibody according to any one of claims 1 to 12, wherein the antibody comprises a human IgG1 or IgG2 heavy chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190 or 192.

19. An isolated antibody according to any one of claims 1 to 12 or 18, wherein the antibody comprises a light chain constant region comprising an amino acid sequence modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions or 1 to 3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO:

194.

20. The isolated antibody according to any one of claims 1 to 19, wherein the antibody comprises a human IgG1 or IgG2 heavy chain constant region comprising the amino acid sequence of any one of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190 or 192.

21. An isolated antibody according to any one of claims 1 to 20, wherein the antibody comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO:

194.

22. An isolated antibody according to any one of claims 1 to 21 which is a full length antibody.

23. An isolated antibody according to any one of claims 1 to 21 which is an IgG antibody lacking a C-terminal lysine in the heavy chain constant region.

24. An isolated antibody according to any one of claims 1 to 12, which is an antibody fragment, such as Fv, single chain Fv (scFv), Fab, Fab' or (Fab')2.

25. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO:68 followed by the amino acid sequence of SEQ ID NO:178; and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:70 followed by the amino acid sequence of SEQ ID NO:

194.

26. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO:68 followed by the amino acid sequence of SEQ ID NO:180; and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:70 followed by the amino acid sequence of SEQ ID NO:

194.

27. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a HC comprising the amino acid sequence of SEQ ID NO: 196 and a light chain comprising the amino acid sequence of SEQ ID NO:

200.

28. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), wherein the antibody comprises: a HC comprising the amino acid sequence of SEQ ID NO: 198 and a light chain comprising the amino acid sequence of SEQ ID NO:

200.

29. An isolated antibody according to any one of claims 1 to 28, which is a bispecific or multispecific antibody, or which is covalently or non-covalently bound to at least one other molecule.

30. The isolated antibody according to claim 29, wherein the antibody is covalently or non-covalently bound to at least one other molecule, wherein the at least one other molecule comprises a detectable label and / or a drug.

31. The isolated antibody according to any one of claims 1 to 30, wherein the antibody does not bind to human protein arginine deiminase 2 (PAD2).

32. The isolated antibody according to any one of claims 1 to 31, wherein the antibody inhibits the conversion of arginine to citrulline in the peptide substrate TSTGGRQGSHH (SEQ ID NO: 216) by PAD4 with an IC50 of 10 to 200 nM, 50 to 200 nM, 10 to 100 nM, 20 to 100 nM or 50 to 100 nM.

33. An isolated antibody according to any one of claims 1 to 32, wherein the antibody optionally inhibits the conversion of arginine to citrulline in a peptide substrate (SHQESTRGKSKGKAAAAA; SEQ ID NO: 232) by PAD4 in a dose-dependent manner with an IC50 of 0.1 to 10 nM (such as 0.2 to 5 nM), wherein the PAD4 concentration is 1 to 8 μg / mL.

34. The isolated antibody according to any one of claims 1 to 33, wherein the antibody inhibits the deamination of N[alpha]-benzoyl-L-arginine ethyl ester hydrochloride (BAEE) by PAD4.

35. according to any one of claims 1 to 34 through the separation of antibodies, wherein the antibody with a K less than 5nM D Binds specifically to PAD4.

36. according to the separated antibody of claim 35, wherein this antibody is with 0.05nM to 0.5nM K D Specific binding to PAD4 is (a) at 37°C in the presence of 1 mM calcium chloride and / or (b) at 37°C in the absence of additional Ca 2+ and measured by surface plasmon resonance (SPR) in the presence of 2 mM EDTA.

37. An isolated antibody according to any one of claims 1 to 36, wherein the antibody has an ECM score of less than 50, less than 30, less than 10, less than 5, 1 to 30, 1 to 20, 1 to 10, 1 to 5, 1 to 3, 1, 2, 3, 4 or 5 in an ECM analysis.

38. An isolated antibody according to any one of claims 1 to 37, wherein the immunogenicity of the antibody is less than one, two or all three of the following as measured by an in silico immunogenicity assay: (alemtuzumab), (rituximab), (daclizumab).

39. The isolated antibody according to any one of claims 1 to 38, wherein the antibody reduces the amount of extracellular citrullinated histone H3 and / or reduces the secretion of GM-CSF and / or the gene expression of GM-CSF in LPS-stimulated human blood monocytes compared to an isotype control antibody.

40. The isolated antibody according to any one of claims 1 to 39, wherein the antibody is capable of being internalized by LPS-stimulated human blood monocytes.

41. An isolated antibody according to any one of claims 1 to 40, wherein the antibody inhibits the function of PAD4 in an inflamed lung, for example as evidenced by reduced citrullination of H3 in bronchoalveolar lavage fluid (BALF) collected from the lung.

42. An isolated antibody according to any one of claims 1 to 41, wherein the antibody inhibits the function of PAD4 in an inflamed joint, for example as evidenced by reduced citrullination of ITIH4 and / or PRG4 in joint tissue.

43. The isolated antibody according to any one of claims 1 to 42, wherein the antibody reduces citrullination of H3 in neutrophils, monocytes, M1 macrophages and / or M2 macrophages compared to an isotype control antibody.

44. An isolated antibody according to any one of claims 1 to 43, wherein the antibody reduces the amount of extracellular citrullinated H3 and / or citrullinated ITIH4 in the BALF in an LPS-induced acute lung inflammation model in human PAD4 knock-in mice, optionally wherein the amount of citrullinated H3 is reduced by at least 15%, at least 30%, at least 40%, at least 50% or at least 60%, and wherein the amount of citrullinated ITIH4 is reduced by at least 50%, at least 70% or at least 80% compared to an isotype control antibody.

45. An isolated antibody according to any one of claims 1 to 44, wherein the antibody reduces the amount of extracellular citrullinated PRG4 and / or citrullinated ITIH4 in mouse patella in an LPS-induced acute joint injury model in human PAD4 gene knock-in mice compared to an isotype control antibody, optionally wherein the EC50 for reducing citrullinated ITIH4 and / or citrullinated PRG4 is 2 nM or less, 1 nM or less, 0.1 nM or less, 0.05 to 2 nM, or 0.1 to 1 nM.

46. ​​A pharmaceutical composition comprising an antibody according to any one of claims 1 to 45 and a pharmaceutically acceptable carrier.

47. An isolated nucleic acid or a collection of two or more nucleic acids encoding an antibody according to any one of claims 1 to 45.

48. An isolated vector comprising one or more nucleic acids encoding the heavy and light chains of an antibody according to any one of claims 1 to 45.

49. An isolated host cell comprising a nucleic acid according to claim 47 or a vector according to claim 48.

50. A method for producing an antibody that specifically binds to PAD4, comprising culturing the host cell according to claim 49 under conditions suitable for expression of the antibody.

51. The method according to claim 50, further comprising recovering the antibody from the host cell.

52. An antibody produced by the method according to claim 50 or 51.

53. Use of an antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46 for the preparation of a medicament for the treatment of an autoimmune disorder.

54. Use according to claim 53, wherein the autoimmune disorder is rheumatoid arthritis, lupus (e.g. systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g. ANCA-associated vasculitis), thrombosis (e.g. venous thrombosis) or inflammatory bowel disease (IBD) (e.g. ulcerative colitis, Crohn's disease).

55. An isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46 for use in treating an autoimmune disorder.

56. The antibody for use according to claim 55, wherein the autoimmune disorder is rheumatoid arthritis, lupus (e.g. systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g. ANCA-associated vasculitis), thrombosis (e.g. venous thrombosis) or inflammatory bowel disease (IBD) (e.g. ulcerative colitis, Crohn's disease).

57. A method of treating an autoimmune disorder in a subject in need thereof, comprising administering to the subject an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

58. The method according to claim 57, wherein the autoimmune disorder is rheumatoid arthritis, lupus (e.g. systemic lupus erythematosus (SLE)), lupus nephritis, vasculitis (e.g. ANCA-associated vasculitis), thrombosis (e.g. venous thrombosis) or inflammatory bowel disease (IBD) (e.g. ulcerative colitis, Crohn's disease).

59. The use, antibody for use or method according to any one of claims 53 to 58, wherein the method or use comprises administering at least one additional therapeutic agent, optionally wherein the at least one additional therapeutic agent is one or more of methotrexate, adalimumab, etanercept, infliximab, hydroxychloroquine, sulfasalazine, leflunomide, abatacept, anakinra, certolizumab, golimumab, rituximab, sarilumab, tocilizumab, baricitinib, tofacitinib or upadacitinib.

60. Use of an antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46 for the preparation of a medicament for treating an individual at risk of developing RA.

61. An isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46 for use in treating a subject at risk of developing RA.

62. A method of treating an individual at risk of developing RA, comprising administering to the individual an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

63. The use, isolated antibody or method according to any one of claims 60 to 62, wherein the subject has one or more of the following conditions: a. At least one first-degree relative (e.g., parent or sibling) with RA; b. The presence of anti-citrullinated protein antibodies (ACPA) in the serum; c. The presence of rheumatoid factor (RF) in serum; d. Arthralgia in at least one joint; e. Inflammation of at least one joint as detected by ultrasound or magnetic resonance imaging (MRI); or f. Undifferentiated arthritis.

64. The use, isolated antibody or method according to any one of claims 60 to 63, wherein the subject is in remission.

65. The use, isolated antibody or method according to any one of claims 60 to 64, wherein the treatment is for delaying the onset of RA.

66. Use of an antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46 for the preparation of a medicament for inhibiting NETosis or METosis in an individual.

67. An isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46 for use in inhibiting NETosis or METosis in an individual.

68. A method of inhibiting NETosis or METosis in a subject, comprising administering to the subject an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

69. A method of inhibiting NETosis in an individual, the method comprising administering to the individual an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

70. A method of inhibiting METosis in a subject, the method comprising administering to the subject an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

71. A method of inhibiting NETosis or METosis in a biological sample in vitro, comprising administering to the biological sample an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

72. A method of inhibiting citrullination in a subject, the method comprising administering to the subject an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

73. A method of inhibiting citrullination in a biological sample in vitro, the method comprising administering to the biological sample an effective amount of an isolated antibody according to any one of claims 1 to 45 or a pharmaceutical composition according to claim 46.

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