Antibodies that bind to human PAD4 and their use

Abstract

This application relates, in particular, to anti-PAD4 (peptidylarginine deiminase 4) antibodies, nucleic acids encoding the antibodies, vectors and host cells containing the nucleic acids, and methods for producing and using the antibodies.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 369,184, filed Jul. 22, 2022, and U.S. Provisional Patent Application No. 63 / 397,698, filed Aug. 12, 2022, the entireties of both of which are incorporated herein by reference.

[0002] Field This application relates to specific anti - PAD4 (peptidylarginine deiminase 4) antibodies, nucleic acids encoding the antibodies, vectors and host cells containing the nucleic acids, and methods of making and using the antibodies.

Background Art

[0003] PAD4 (peptidylarginine deiminase 4) is one of five PAD enzymes (PAD1, 2, 3, 4, and 6) in humans, is found in some immune cell types, and acts by converting arginine to citrulline (Chavanas et al., Gene 330:19-27 (2004).). Such 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, in which neutrophils extrude a complex of decondensed chromatin structures 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). Moreover, proteins citrullinated by PAD4 become antigenic substrates and are targets of both 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.). Thus, PAD4 activity can lead to the production of anti-citrullinated protein antibodies (ACPA).

[0004] PAD4 has been suggested to play a role in diseases such as rheumatoid arthritis (RA), lupus (including systemic lupus erythematosus (SLE), lupus nephritis), vasculitis (including antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis), inflammatory bowel disease (IBD) (including ulcerative colitis and Crohn's disease), thrombosis (e.g., venous thrombosis), antiphospholipid antibody syndrome, cystic fibrosis, and cancer, etc. (see, for example, Curran et al.; Yadav et al., J. Cyst. Fibros. 18:636-45 (2018); Wang et al., Front. Immunol. 13:895216 (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., Authorea 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 with both tissue (i.e., joint and non-joint) and systemic inflammation. The progression of RA is associated with an increase in autoantibody levels, and approximately 70% of RA patients are positive for anti-citrullinated protein antibodies (ACPA). ACPA appears in the lungs and blood of people at risk of RA, and high ACPA levels are associated with severe RA symptoms such as bone destruction and certain comorbidities, such as cardiovascular disease. In addition, the presence of ACPA worsened RA symptoms in a murine spontaneous arthritis model.

[0005] ACPA recognizes neoantigens, particularly those from proteins citrullinated by PAD enzymes. PAD4 is the major PAD enzyme found in synovial tissue. For example, PAD4 can mediate ACPA-dependent and ACPA-independent functions important for the development of RA and other diseases. In addition, approximately 15% of RA patients have antibodies that activate PAD4. The presence of such PAD4-activating antibodies correlates with severe joint-destructive disease. At the gene level, single nucleotide polymorphisms (SNPs) in the PADI4 gene encoding PAD4 that contribute to RA susceptibility haplotypes have been identified (Suzuki et al., Nat. Genet. 34:395-402 (2003)). Furthermore, epigenetic changes in the promoter region of PADI4 are associated with disease activity with lower methylation in RA patients compared to healthy individuals (Kolarz et al., Clin. Med. 9(7):2049 (2020)). RA patients with susceptibility haplotypes for PADI4 have been found to have higher titers of anti-citrullinated peptide antibodies (ACPA) than RA patients without the haplotype (Reyes-Castillo et al., Clin. Exp. Immunol. 182(2):119-31 (2015)).

[0006] Accordingly, blocking the activity of PAD4 may be useful in the treatment of RA, and for treating subjects at risk of developing RA, as well as in the treatment of other inflammatory and autoimmune diseases, or diseases associated with NETosis, METosis, the presence of anti-citrullinated protein antibodies (ACPA), increased PAD4 expression, or increased PAD4 activity, such as increased citrullination of polypeptides.

[0007] The present disclosure provides anti-PAD4 antibodies that inhibit the activity of PAD4, nucleic acids encoding the antibodies, vectors and host cells comprising the antibodies, and methods of making and using the antibodies.

Summary of the Invention

Problems to be Solved by the Invention

[0008] The present disclosure relates in particular to anti-PAD4 antibodies, nucleic acids encoding the antibodies, vectors and host cells comprising the antibodies, and methods of making and using the antibodies. For example, embodiments of the present disclosure include the following.

[0009] 1. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO: 4 or 62, and an HCDR3 having the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO: 7.

[0010] 2. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO: 4 or 62, an HCDR2 having the amino acid sequence of SEQ ID NO: 5, and an HCDR3 having the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO: 7, an LCDR2 having the amino acid sequence of SEQ ID NO: 8, and an LCDR3 having the amino acid sequence of SEQ ID NO: 9.

[0011] 3. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) having the amino acid sequences of SEQ ID NO: 221 or 225 and SEQ ID NO: 222; and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO: 223.

[0012] 4. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) having 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) having the amino acid sequence of SEQ ID NO: 223, further comprising a light chain constant region having the amino acid sequence of SEQ ID NO: 224 or 235.

[0013] 5. An isolated antibody according to any one of embodiments 1 to 4, comprising a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to any one of the amino acid sequences of SEQ ID NO: 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 any one of the amino acid sequences of SEQ ID NO: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70.

[0014] 6. An isolated antibody according to any one of embodiments 1 to 4, comprising a VH comprising any one of the amino acid sequences of SEQ ID NO: 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; and a VL comprising any one of the amino acid sequences 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.

[0015] 7. An isolated antibody according to any one of embodiments 1 to 6, wherein the heavy chain variable region comprises glycine at Kabat position 94 (corresponding to position 98 of SEQ ID NO: 10).

[0016] An isolated antibody according to any one of embodiments 1-7, comprising a VH comprising any one amino acid sequence of SEQ ID NOs: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68.

[0017] An isolated antibody according to any one of embodiments 1-8, comprising a VL comprising any one amino acid sequence of SEQ ID NOs: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70.

[0018] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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.

[0019] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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.

[0020] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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.

[0021] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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.

[0022] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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.

[0023] 15. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated 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.

[0024] 16. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0025] 17. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0026] 18. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0027] 19. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0028] 20. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0029] 21. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0030] 22. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0031] 23. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0032] 24. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0033] 25. The isolated antibody according to any one of embodiments 1 to 23, which binds to an epitope on PAD4 comprising SEQ ID NO: 217 and SEQ ID NO: 218.

[0034] 26. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 72, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 73, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 74; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 75, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 76, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 77.

[0035] An isolated antibody according to embodiment 26, comprising a VH that is at least 90%, at least 95%, at least 97%, or at least 99% identical to any one of the amino acid sequences of SEQ ID NO: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134; and a VL that is at least 90%, at least 95%, at least 97%, or at least 99% identical to any one of the amino acid sequences of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136.

[0036] An isolated antibody according to embodiment 26, comprising a VH comprising any one of the amino acid sequences of SEQ ID NO: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134, which is 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 any one of the amino acid sequences of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136, which is modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions.

[0037] An isolated antibody according to embodiment 25, 26, or 27, comprising a VH comprising any one of the amino acid sequences of SEQ ID NO: 78, 82, 86, 90, 94, 98, 102, 106, 110, 114, 118, 122, 126, 130, or 134.

[0038] An isolated antibody according to any one of embodiments 26 - 29, comprising a VL comprising any one of the amino acid sequences of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136.

[0039] 31. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0040] 32. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0041] 33. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0042] 34. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0043] 35. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0044] 36. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0045] 37. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0046] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0047] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0048] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0049] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0050] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0051] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0052] An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0053] 45. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a VH containing the amino acid sequence of SEQ ID NO: 134 and a VL containing the amino acid sequence of SEQ ID NO: 136.

[0054] 46. The isolated antibody according to any one of embodiments 26 to 45, which binds to an epitope on PAD4 comprising SEQ ID NO: 219 and SEQ ID NO: 220.

[0055] 47. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), a. Heavy chain complementarity-determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 138, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 138, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 138; b. Heavy chain complementarity-determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 140, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 140; c. Heavy chain complementarity-determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 142, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 142, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 142; d. Heavy chain complementarity-determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 144, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 144, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 144; e. Heavy chain complementarity-determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 146, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 146, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 146; f. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 148, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 148, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 148; g. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 150, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 150, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 150; h. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 152, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 152, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 152; i. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 154, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 154, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 154; j. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 156, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 156, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 156; k. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 158, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 158, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 158; l. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 160, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 160, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 160; m. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 162, HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 162, and HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 162; n. Heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 164, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 164, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 164; o. Heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 166, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 166, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 166; p. Heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 168, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 168, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 168; or q. Heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 170, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 170, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 170 comprising a heavy chain variable region (VH); and a light chain variable region (VL) comprising light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 170, LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and LCDR3 comprising the amino acid sequence of residues 93-101 of SEQ ID NO: 170, an isolated antibody.

[0056] 48. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), a. Heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 138, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 138, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 138; b. Heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 140, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 140, and HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 140; c. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 142, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 142, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 142; d. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 144, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 144, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 144; e. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 146, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 146, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 146; f. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 148, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 148, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 148; g. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 150, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 150, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 150; h. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 152, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 152, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 152; i. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 154, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 154, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 154; j. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 156, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 156, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 156; k. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 158, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 158, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 158; l. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 160, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 160, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 160; m. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 162, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 162, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 162; n. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 164, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 164, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 164; o. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 166, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 166, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 166; or p. Heavy chain complementarity-determining region 1 (HCDR1) containing the amino acid sequence at positions 26-35 of SEQ ID NO: 168, HCDR2 containing the amino acid sequence at positions 50-66 of SEQ ID NO: 168, and HCDR3 containing the amino acid sequence at positions 99-108 of SEQ ID NO: 168; comprising a heavy chain variable region (VH) comprising; and a light chain variable region (VL) comprising a light chain complementarity-determining region 1 (LCDR1) containing the amino acid sequence of residues 24-38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54-60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93-101 of SEQ ID NO: 172, an isolated antibody.

[0057] 49. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) containing a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 168, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 168, and an HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 168; and a light chain variable region (VL) containing a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 172, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 172, and an LCDR3 comprising the amino acid sequence of residues 93-101 of SEQ ID NO: 172, the isolated antibody.

[0058] 50. The isolated antibody according to embodiment 49, which binds to PAD4 at pH 6.0 with a 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.

[0059] 51. The isolated antibody according to any one of embodiments 1-50, which is an IgA, IgG, or IgM antibody.

[0060] 52. The isolated antibody according to any one of embodiments 1-50, which is an IgG antibody, such as a human IgG1, IgG2, IgG3, or IgG4 antibody, or a mouse IgG1 or IgG2 antibody.

[0061] 53. The isolated antibody according to embodiment 52, which comprises a wild-type, human IgG1, IgG2, or IgG4 heavy chain constant region.

[0062] An isolated antibody according to any one of embodiments 1 to 50, comprising a human IgG1 or IgG2 heavy chain constant region that is at least 90%, at least 95%, at least 97%, or at least 99% identical to any one of the amino acid sequences of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and / or comprising a light chain constant region 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.

[0063] An isolated antibody according to any one of embodiments 1 to 50, comprising a human IgG1 or IgG2 heavy chain constant region modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions compared to any one of the amino acid sequences of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and / or comprising a light chain constant region 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.

[0064] An isolated antibody according to any one of embodiments 1 to 55, comprising a human IgG1 or IgG2 heavy chain constant region comprising any one of the amino acid sequences of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192.

[0065] An isolated antibody according to any one of embodiments 1 to 56, comprising a light chain constant region comprising the amino acid sequence of SEQ ID NO: 194.

[0066] An isolated antibody according to any one of embodiments 1 to 57, which is a full-length antibody.

[0067] An isolated antibody according to any one of embodiments 1 to 57, which is an IgG antibody lacking a C-terminal lysine in the heavy chain constant region.

[0068] 60. An isolated antibody according to any one of embodiments 1 to 50, which is an antibody fragment, such as Fv, single-chain Fv (scFv), Fab, Fab', or (Fab')2.

[0069] 61. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 68 and the subsequent amino acid sequence of SEQ ID NO: 178, and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 70 and the subsequent amino acid sequence of SEQ ID NO: 194.

[0070] 62. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 68 and the subsequent amino acid sequence of SEQ ID NO: 180, and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 70 and the subsequent amino acid sequence of SEQ ID NO: 194.

[0071] 63. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising an 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.

[0072] 64. An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising an 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.

[0073] 65. An isolated antibody according to any one of embodiments 1 to 64, which is a bispecific or multispecific antibody, or is conjugated to at least one other molecule by a covalent or non-covalent bond.

[0074] 66. An isolated antibody according to embodiment 65, which is conjugated by covalent or non-covalent bonds to at least one other molecule, and wherein the at least one other molecule comprises a detection label and / or a drug.

[0075] 67. An isolated antibody that specifically binds to mouse protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 31-35 of SEQ ID NO: 208, an HCDR2 comprising the amino acid sequence at positions 50-56 of SEQ ID NO: 208, and an HCDR3 comprising the amino acid sequence at positions 99-107 of SEQ ID NO: 208; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 210, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 210, and an LCDR3 comprising the amino acid sequence of residues 93-101 of SEQ ID NO: 210.

[0076] 68. An isolated antibody according to embodiment 67, comprising a human IgG1 heavy chain variable region (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 / or 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.

[0077] 69. An isolated antibody according to embodiment 67, comprising a VH amino acid sequence modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions compared to any one of the amino acid sequences of SEQ ID NO: 208; and / or a VL amino acid sequence modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 210.

[0078] 70. An isolated antibody according to any one of embodiments 67-69, comprising the VH amino acid sequence of SEQ ID NO: 208 and / or the VL amino acid sequence of SEQ ID NO: 210.

[0079] 71. An isolated antibody according to any one of embodiments 67-70, comprising 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 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.

[0080] 72. An isolated antibody according to any one of embodiments 67-71, comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 212, and / or a light chain comprising the amino acid sequence of SEQ ID NO: 214.

[0081] 73. An isolated antibody according to any one of embodiments 1-66 that does not bind to human protein arginine deiminase 2 (PAD2).

[0082] 74. An isolated antibody according to any one of embodiments 1-66 or 73 that inhibits the conversion of arginine to citrulline by PAD4 in the peptide substrate TSTGGRQGSHH (SEQ ID NO: 216) with an IC50 of 10-200 nM, 50-200 nM, 10-100 nM, 20-100 nM, or 50-100 nM.

[0083] 75. An isolated antibody according to any one of embodiments 1-66, 73, or 74 that dose-dependently inhibits the conversion of arginine to citrulline by PAD4 in the peptide substrate (SHQESTRGKSKGKAAAAA; SEQ ID NO: 232) with an IC50 of 0.1-10 nM, for example 0.2-5 nM, at an appropriate PAD4 concentration of 1-8 μg / mL.

[0084] An isolated antibody according to any one of embodiments 1-66 or 73-75 that inhibits PAD4 deamination of Nα-benzoyl-L-arginine ethyl ester hydrochloride (BAEE).

[0085] 77. An isolated antibody according to any one of embodiments 1-66 or 73-76 that specifically binds to PAD4 with a K of less than 77.5 nM, less than 3 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, from 0.01 nM to 5 nM, from 0.01 nM to 1 nM, from 0.05 nM to 1 nM, from 0.1 nM to 1 nM, from 0.1 nM to 0.5 nM, from 0.05 nM to 0.5 nM, from 0.05 nM to 0.1 nM, or from 0.5 nM to 1 nM. D

[0086] 78. An isolated antibody according to embodiment 77 that specifically binds to PAD4 with 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, from 0.01 nM to 5 nM, from 0.01 nM to 1 nM, from 0.05 nM to 1 nM, from 0.1 nM to 1 nM, from 0.1 nM to 0.5 nM, from 0.05 nM to 0.5 nM, from 0.05 nM to 0.1 nM, or from 0.5 nM to 1 nM as determined by surface plasmon resonance (SPR) (a) in the presence of 1-2 mM calcium chloride (e.g., 1 mM calcium chloride) at 37°C and / or (b) in the absence of added Ca and in the presence of 2 mM EDTA at 37°C. 2+ D

[0087] 79. An isolated antibody according to any one of embodiments 1-66 or 73-78 that has an ECM score of less than 50, less than 30, less than 10, less than 5, from 1 to 30, from 1 to 20, from 1 to 10, from 1 to 5, from 1 to 3, 1, 2, 3, 4, or 5 in an ECM assay.

[0088] An isolated antibody according to any one of embodiments 1-66 or 73-79, which is less immunogenic than one, two, or all three of Campath® (alemtuzumab), Rituxan® (rituximab), and Zenapax® (daclizumab) when measured in an in silico immunogenicity assay.

[0089] An isolated antibody according to any one of embodiments 1-66 or 73-80, which reduces the amount of extracellular citrullinated histone H3 in LPS-stimulated human blood monocytes and / or reduces the secretion of GM-CSF and / or the gene expression of GM-CSF as compared to an isotype control antibody.

[0090] An isolated antibody according to any one of embodiments 1-66 or 73-81, which is capable of internalizing by LPS-stimulated human blood monocytes.

[0091] An isolated antibody according to any one of embodiments 1-82, which inhibits PAD4 function in an inflamed lung, as demonstrated by a decrease in citrullination (e.g., of H3) in bronchoalveolar lavage fluid (BALF) collected from the lung.

[0092] An isolated antibody according to any one of embodiments 1-83, which inhibits PAD4 function in an inflamed joint, as demonstrated by a decrease in citrullination (e.g., of ITIH4 and / or PRG4) in joint tissue.

[0093] An isolated antibody according to any one of embodiments 1-84, which reduces citrullination (e.g., of H3) in neutrophils, monocytes, M1 macrophages, and / or M2 macrophages as compared to an isotype control antibody.

[0094] 86. The amount of extracellular citrullination (e.g., of H3 and / or ITIH4) in BALF in an LPS-induced acute lung inflammation model using human PAD4 knock-in mice is reduced as compared to an isotype control antibody, and the amount of citrullinated H3 is reduced by at least 15%, at least 30%, at least 40%, at least 50%, or at least 60% as appropriate, and / or the amount of citrullinated ITIH4 is reduced by at least 50%, at least 70%, or at least 80%, the isolated antibody according to any one of Embodiments 1 to 66 or 73 to 85.

[0095] 87. The amount of extracellular citrullination (e.g., of PRG4 and / or ITIH4) in the mouse patella in an LPS-induced acute joint injury model using human PAD4 knock-in mice is reduced as compared to an isotype control antibody, and the EC50 of the reduction of 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 as appropriate, the isolated antibody according to any one of Embodiments 1 to 66 or 73 to 86.

[0096] 88. A pharmaceutical composition comprising the antibody according to any one of Embodiments 1 to 87 and a pharmaceutically acceptable carrier.

[0097] 89. An isolated nucleic acid or a set of two or more nucleic acids encoding the antibody according to any one of Embodiments 1 to 87.

[0098] 90. An isolated vector comprising one or more nucleic acids encoding the heavy chain and the light chain of the antibody according to any one of Embodiments 1 to 87.

[0099] 91. An isolated host cell comprising the nucleic acid according to Embodiment 89 or the vector according to Embodiment 90.

[0100] 92. A method for producing an antibody that specifically binds to PAD4, comprising culturing the host cell according to Embodiment 91 under conditions suitable for the expression of the antibody.

[0101] The method according to embodiment 92, further comprising recovering the antibody from the host cell.

[0102] An antibody produced by the method of embodiment 92 or 93.

[0103] Use of the antibody according to any one of embodiments 1 to 87, or the pharmaceutical composition according to embodiment 88, in the preparation of a medicament for treating an autoimmune disease.

[0104] 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.

[0105] 97. An isolated antibody according to any one of embodiments 1 to 87 or a pharmaceutical composition according to embodiment 88 for use in the treatment of an autoimmune disorder.

[0106] 98. The antibody for use according to 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.

[0107] 99. A method for 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 embodiments 1 to 87 or a pharmaceutical composition according to embodiment 88.

[0108] 100. The method according to 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.

[0109] 101. The use, antibody for use, or method according to any one of embodiments 95 to 100, comprising 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.

[0110] Further exemplary embodiments include the antibody according to any one of embodiments 1 to 87, comprising at least one post-translational modification of the amino acid sequence. In some embodiments, the modification is the 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.

[0111] The disclosure of this specification also includes the use of an antibody described in any one of Embodiments 1 to 87 above or the pharmaceutical composition described in Embodiment 88 above in the preparation of a medicament for treating a subject at risk of developing RA, or the use of an isolated antibody described in any one of Embodiments 1 to 87 above or the pharmaceutical composition of Embodiment 88 above for use in treating a subject at risk of developing RA. The disclosure also includes a method of treating a subject at risk of developing RA, the method comprising administering to the subject an effective amount of an isolated antibody of any one of Embodiments 1 to 87 above or the pharmaceutical composition of Embodiment 88. In some examples, 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 the serum; (d) joint pain in at least one joint; (e) the presence of inflammation in at least one joint observed by ultrasound or magnetic resonance imaging (MRI); or (f) undifferentiated arthritis. In some examples, the subject is in remission. In some examples, the method or use includes administering at least one additional therapeutic agent, and optionally 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.

[0112] In a further embodiment, the antibody described in any one of Embodiments 1 to 87 or the pharmaceutical composition described in Embodiment 88 may be used in the preparation of a medicament for inhibiting NETosis or METosis in a subject. In some embodiments, the subject is a subject having 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, such as rheumatoid arthritis. Additional embodiments include an isolated antibody described in any one of Embodiments 1 to 87 or the pharmaceutical composition described in Claim 88 for use in inhibiting NETosis or METosis in a subject. In some embodiments, the subject is a subject having 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 a subject at risk of developing an autoimmune disorder, such as rheumatoid arthritis. Further embodiments also include a method of inhibiting NETosis or METosis in a subject, the method comprising administering to the subject an effective amount of the antibody described in any one of Embodiments 1 to 87 or the pharmaceutical composition described in Embodiment 88. In some embodiments, the subject is a subject having 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 subject is a subject at risk of developing an autoimmune disorder, such as rheumatoid arthritis.In some examples, the method or use includes administering at least one additional therapeutic agent, and optionally, 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. The present disclosure herein also encompasses an in vitro method of inhibiting NETosis or METosis in a biological sample, the method comprising administering to the biological sample an effective amount of the isolated antibody according to any one of Embodiments 1 to 87 or the pharmaceutical composition according to Embodiment 88. The methods and uses herein also include a method of inhibiting citrullination in a subject, the method comprising administering to the subject an effective amount of the isolated antibody according to any one of Embodiments 1 to 87, or the use of the isolated antibody according to any one of Embodiments 1 to 87 for inhibiting citrullination in a subject, or the use of the 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 disclosure also includes an in vitro method of inhibiting citrullination in a biological sample, the method comprising administering to the biological sample an effective amount of the isolated antibody according to any one of Embodiments 1 to 87.

[0113] In a further embodiment, the antibody described in any one of Embodiments 1 to 87 or the pharmaceutical composition described in Embodiment 88 may be used for the preparation of a medicament for preventing the occurrence 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, or the isolated antibody described in any one of Embodiments 1 to 87 or the pharmaceutical composition described in Claim 88 may be for use in preventing the occurrence 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. A still further embodiment includes a method of preventing the occurrence 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, the method comprising administering to the subject an effective amount of the isolated antibody described in any one of Embodiments 1 to 87 or the pharmaceutical composition described in Claim 88. In some embodiments, the subject has been determined to be susceptible to the occurrence or recurrence of the disorder. In some such uses and methods herein, the antibody is administered to the subject together with an additional therapeutic agent.

[0114] In additional embodiments, the isolated antibody described in any one of Embodiments 1-87 or the pharmaceutical composition described in Embodiment 88 may be used for the preparation of a medicament for treating cancer, or the isolated antibody or pharmaceutical composition may be for use in treating cancer. Further embodiments also include a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of the isolated antibody described in any one of Embodiments 1-87 or the pharmaceutical composition described in Embodiment 88. In some examples, such a method or use comprises administering to the subject at least one additional therapeutic agent, such as an immune checkpoint inhibitor, a chemotherapeutic agent, an anti-angiogenic agent, or an anti-neoplastic agent, or additional treatments such as radiation therapy, hormone therapy, or surgical procedures.

[0115] In further embodiments, the antibody described in any one of Embodiments 1-87 or the pharmaceutical composition described in Embodiment 88 may be used for the preparation of a medicament for treating an infectious disease, or the isolated antibody described in any one of Embodiments 1-87 or the pharmaceutical composition described in Claim 88 may be for use in treating an infectious disease. Still further embodiments include a method of treating an infectious disease in a subject in need thereof, the method comprising administering an effective amount of the antibody described herein. In some such uses and methods herein, the antibody is administered with an additional therapeutic agent.

[0116] The foregoing general description and the following detailed description are exemplary and explanatory only and are not to be construed as limiting the claims. All references cited herein are hereby incorporated by reference in their entirety. BRIEF DESCRIPTION OF THE DRAWINGS

[0117]

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Modes for Carrying Out the Invention

[0118] I. Definitions Unless otherwise defined, scientific and technical terms used in connection with the present invention have the meanings commonly understood by those skilled in the art. Further, unless the context specifically requires otherwise, terms in the singular form include the plural, and terms in the plural form include the singular.

[0119] In the present application, the use of "or" means "and / or" unless otherwise stated. In the context of multiple dependent claims, the use of "or" is limited to alternatives and refers back to one or more preceding independent or dependent claims. Similarly, the terms "element" or "component" include both elements and components that include one unit and elements and components that include more than one subunit, unless otherwise stated.

[0120] Exemplary techniques used in connection 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, N.Y. (1989)).

[0121] As used herein, the term "about" refers to numerical values, including, for example, integers, decimals, and percentages, whether or not explicitly indicated. The term "about" generally refers to a range of numerical values (e.g., + / - 5 to 10% of the recited range) that a person of ordinary skill in the art would consider equivalent to the recited value (e.g., having the same function or result). When terms such as at least and about precede a list of numerical values or ranges, the terms modify all of the values or ranges provided in the list. In some instances, the term about may include numerical values rounded to the nearest significant digit.

[0122] The term "polypeptide" refers to a polymer of amino acid residues and is not limited to a minimum length. A "protein" may contain one or more polypeptides. Such polymers of amino acid residues may contain, but are not limited to, natural or non-natural amino acid residues and include peptides, oligopeptides, dimers, trimers, and multimers of amino acid residues. Both full-length proteins and fragments thereof are included in the definition. The term also includes post-expression modifications of polypeptides, such as glycosylation, sialylation, acetylation, phosphorylation, etc. Further, for the purposes of the present invention, "polypeptide" or "protein" each refers to a polypeptide or protein that includes modifications such as deletions, additions, and substitutions (generally, essentially conservative) with respect to the native sequence, 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 that gives rise to the protein or errors during PCR amplification. A protein may include two or more polypeptides.

[0123] "PAD4" or "protein arginine deiminase 4" or "peptidyl arginine deiminase 4", as used herein, refers to human PAD4 (huPAD4; UniProt ID: Q9UM07) unless expressly stated otherwise (i.e., mouse PAD4, cynomolgus monkey 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.

[0124] The term "antibody" refers to a molecule that includes at least complementarity determining regions (CDRs) 1, 2, and 3 of the heavy chain and at least CDRs 1, 2, and 3 of the light chain and is capable of binding to an antigen. This term is used in the broadest sense and includes, but is not limited to, various antibody structures such as 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.

[0125] An "isolated" antibody is one that has been separated from the components of its natural environment. In some embodiments, the antibody is purified to a purity greater than 95% or 98% as determined by, for example, electrophoretic (e.g., SDS-PAGE, isoelectric focusing electrophoresis (IEF), capillary electrophoresis) or chromatographic (e.g., ion exchange or reverse phase HPLC) methods. For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[0126] "Antigen" refers to the target of an antibody, i.e., the molecule to which the antibody specifically binds. The term "epitope" refers to the site on a protein-like or non-protein-like antigen to which an antibody binds. Epitopes on a protein can be formed from either a continuous stretch of amino acids (linear epitope) or from non-contiguous amino acids that are spatially proximate, e.g., by the folding of the antigen, i.e., the three-dimensional folding of a protein-like antigen (conformational epitope). Linear epitopes typically remain bound to an antibody even after the protein-like antigen has been exposed to a denaturing agent, whereas conformational epitopes are typically disrupted by treatment with a denaturing agent.

[0127] "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.

[0128] The term "heavy chain" refers to a polypeptide that includes at least a heavy chain variable region, with or without a leader sequence. In some embodiments, the heavy chain includes at least a portion of the heavy chain constant region. The term "full-length heavy chain" refers to a polypeptide that includes a heavy chain variable region and a heavy chain constant region, with or without a leader sequence.

[0129] The term "light chain" refers to a polypeptide that includes at least a light chain variable region, with or without a leader sequence. In some embodiments, the light chain includes at least a portion of the light chain constant region. The term "full-length light chain" refers to a polypeptide that includes a light chain variable region and a light chain constant region, with or without a leader sequence.

[0130] "Complementary determining region" ("CDR"), as used herein, refers to each of the regions of the antibody variable region where the sequence is highly variable and determines antigen-binding specificity. Generally, an antibody includes six CDRs: three in VH (CDR-H1 or heavy chain CDR1, CDR-H2, CDR-H3) and three in VL (CDR-L1, CDR-L2, CDR-L3). Unless otherwise indicated, CDRs are determined according to the sequence listing herein.

[0131] "Framework" or "FR" refers to the residues of the variable region residues that are not part of the complementary determining region (CDR). The FRs of the variable region generally consist of four FRs: FR1, FR2, FR3, and FR4. Thus, CDR and FR sequences generally 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.

[0132] The term "variable region" or "variable domain" refers to the domain of the antibody heavy or light chain that is involved in binding of the antibody to an antigen. The variable domains of the heavy and light chains of a native antibody (VH and VL, respectively) generally have a similar structure, and each domain includes four conserved framework regions (FRs) and three complementary determining regions (CDRs). For example, Kindt et al. Kuby Immunology, 6 thSee, e.g., ed., W.H. Freeman and Co., page 91 (2007). The variable domain includes heavy chain (HC) CDR1-FR2-CDR2-FR3-CDR3, with or without all or part of FR1 and / or FR4; and light chain (LC) CDR1-FR2-CDR2-FR3-CDR3, with or without all or part of FR1 and / or FR4. That is, the variable domain may lack part 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. Further, an antibody that binds a particular antigen may be isolated using a VH or VL domain from an antibody that binds the antigen and screening a library 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).

[0133] The "constant regions" of the light and heavy chains of an antibody refer to additional sequence portions outside of the FRs, CDRs, and variable regions. Certain antibody fragments may lack all or part of the constant regions. Each heavy chain has, from N- to C-terminus, three heavy chain constant domains (CH1, CH2, and CH3) following the variable domain, also called the variable heavy chain domain or heavy chain variable region (VH). Similarly, each light chain has, from N- to C-terminus, a light chain constant (CL) domain following the variable domain, also called the light chain variable domain or light chain variable region (VL).

[0134] The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the 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 Pro230 to the carboxyl terminus Gly446 and Lys447 (EU numbering) of the heavy chain. Antibodies produced by a host cell may undergo post-translational cleavage of one or more, particularly one or two, amino acids from the C-terminus of the heavy chain. Thus, an antibody produced by a host cell upon expression of a particular nucleic acid molecule encoding a full-length heavy chain may contain the full-length heavy chain or a cleavage variant of the full-length heavy chain. This can be the case when the last two C-terminal amino acids of the heavy chain are glycine and lysine, respectively. Thus, the C-terminal lysine, or the C-terminal glycine and lysine, of the Fc region may or may not be present. Thus, for example, a "full-length heavy chain constant region" or "full-length antibody" that is a human IgG1 antibody includes IgG1 having both C-terminal glycine and lysine, those having no C-terminal lysine, or those having neither C-terminal glycine and lysine. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region follows EU numbering, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5 th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991, and is referred to as EU numbering.

[0135] "Effector function" refers to those biological activities attributable to the Fc region of an antibody that vary by 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 receptors); and B cell activation.

[0136] The "class" of an antibody refers to the type of constant domain or constant region carried by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of these may be further classified 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, β, ε, γ, and μ, respectively. The light chain of an antibody can be assigned to one of two types called kappa (κ) and lambda (λ) based on the amino acid sequence of its constant domain.

[0137] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to an antigen (i.e., PAD4) to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab’, Fab’-SH, F(ab’)2; diabody; linear antibody; single-chain antibody molecule (e.g., scFv, and scFab); single domain antibody (dAbs); and multispecific antibodies formed from antibody fragments. For an assessment of certain antibody fragments, see Holliger and Hudson, Nature Biotechnology 23:1126-1136 (2005).

[0138] The terms "full-length antibody", "intact antibody", and "whole antibody" are used interchangeably herein and refer to an antibody having a structure substantially similar to the native antibody structure, or, in the case of an IgG antibody, an antibody having a heavy chain containing the Fc region as defined above herein.

[0139] The term "chimeric" antibody refers to an antibody in which a portion of the heavy chain and / or light chain is derived from a particular origin or species, but the remainder of the heavy chain and / or light chain is derived from a different origin or species.

[0140] A "humanized" antibody refers to a chimeric antibody that contains amino acid residues from non-human CDRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody comprises substantially all of at least one, typically two, variable domains in which all or substantially all of the CDRs correspond to those of a non-human antibody and all or substantially all of the FRs correspond to those of a human antibody. A humanized antibody may comprise at least a portion of an antibody constant region derived from a human antibody. An "humanized form" of an antibody, e.g., a non-human antibody, refers to an antibody that has been humanized.

[0141] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical and / or bind the same epitope, except for possible variants that are present in minor amounts, typically containing naturally occurring mutations or mutations that occur during production of the monoclonal antibody preparation. In contrast to polyclonal antibody preparations that typically include different antibodies against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation binds to a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method.

[0142] A "bispecific" antibody is an antibody that specifically binds to more than one target antigen, and a "bispecific" antibody is an antibody that specifically binds to two antigens. An "antibody conjugate" is an antibody conjugated to one or more heterologous molecules, including, but not limited to, a therapeutic agent or a label.

[0143] Antibodies can be modified as part of the production process through metabolism in a particular host cell or in vivo. The amino acid sequences of the antibodies or antibody regions herein are intended to include not only specific amino acid sequences but also post-translationally modified sequences including, for example, side chain modifications and cleavages. Such post-translational modifications can occur, for example, as a result of antibody production in a host cell and / or as a result of post-translational modification in vivo in an animal (e.g., a human).

[0144] In some embodiments, the antibodies disclosed herein include post-translational modifications (e.g., one or more post-translational modifications). Post-translational modifications can include, for example, ubiquitination, phosphorylation, acetylation, hydroxylation, methylation, glycosylation, AMPylation, prenylation, deamidation, elimylation, citrullination, and carbamoylation. In some embodiments, the antibody is not post-translationally modified.

[0145] As noted above, antibodies can undergo post-translational cleavage of one or more, particularly one or two, amino acids, often Gly-Lys, from the C-terminus of the heavy chain. This cleavage can occur, for example, as a result of the antibody production process in a host cell. Antibodies produced by expression of a particular nucleic acid molecule encoding a full-length heavy chain can include the full-length heavy chain or a cleaved variant of the full-length heavy chain, e.g., a heavy chain lacking the C-terminal Lys or C-terminal Gly-Lys.

[0146] Other types of post-translational modifications can occur during antibody production or otherwise in vivo, e.g., during modification of amino acid side chains. For example, the N-terminal Glu or Gln residue in an antibody chain can be post-translationally modified to N-terminal pyroglutamate (also known as pyrrolidine carboxylate; the abbreviation pE).

[0147] "Percent (%) amino acid sequence identity" and "homology" with respect to a peptide, polypeptide, or antibody sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a specific peptide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignments for the purpose of determining percent amino acid sequence can be achieved in a variety of ways within the skill of the art, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGNTM (DNASTAR) software. One of ordinary skill in the art can determine appropriate parameters for measuring the alignment, including any algorithms necessary to achieve the maximum alignment over the full length of the sequences being compared.

[0148] The terms "signal sequence" or "leader sequence" refer to the sequence of amino acid residues located at the N-terminus of a polypeptide that facilitates the secretion of the polypeptide from mammalian cells. The leader sequence can be cleaved when the polypeptide is exported from mammalian cells to form the mature protein. The leader sequence can be natural or synthetic, and they can be heterologous or homologous to the protein to which they are 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 selected from a natural leader sequence and a heterologous leader sequence.

[0149] The term "nucleic acid molecule" or "polynucleotide" includes any compound and / or substance that comprises a polymer of nucleotides. Each nucleotide is composed of a base, specifically a purine or pyrimidine base (i.e., cytosine (C), guanine (G), adenine (A), thymine (T) or uracil (U)), a sugar (i.e., deoxyribose or ribose), and a phosphate group. Often, nucleic acid molecules are described by the sequence of bases, which represents the primary structure of the nucleic acid molecule (linear structure). The sequence of bases is typically represented 5' to 3'. As used herein, the term nucleic acid molecule includes deoxyribonucleic acid (DNA), including, for example, complementary DNA (cDNA) and genomic DNA, ribonucleic acid (RNA), particularly messenger RNA (mRNA), synthetic forms of DNA or RNA, and hybrid polymers containing two or more of these molecules. Nucleic acid molecules can be linear or circular. Additionally, the term nucleic acid molecule includes both sense and antisense strands, as well as single-stranded and double-stranded forms. Moreover, the nucleic acid molecules described herein can contain nucleotides that are naturally occurring or non-naturally occurring. Examples of non-naturally occurring nucleotides include modified nucleotide bases having derivatized sugars or phosphate backbone linkages or chemically modified residues. Nucleic acid molecules also include, for example, DNA and RNA molecules suitable as vectors for the in vitro and / or in vivo direct expression of the antibodies of the present invention in a host or patient. Such DNA (e.g., cDNA) or RNA (e.g., mRNA, circular RNA) vectors can be unmodified or modified.

[0150] "Isolated" nucleic acids refer to nucleic acid molecules that are separated from the components of their natural environment. Isolated nucleic acids typically include nucleic acid molecules contained in cells that contain the nucleic acid molecule, but the nucleic acid molecule is present at an extrachromosomal location or at a chromosomal location different from its location on the natural chromosome.

[0151] "Isolated nucleic acid encoding an anti-PAD4 antibody" refers to one or more nucleic acid molecules encoding an anti-PAD4 antibody heavy chain and light chain (or fragments thereof), such nucleic acid molecules in a single vector or separate vectors, and such nucleic acid molecules present at one or more locations in a host cell.

[0152] 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 as 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 operably linked. Such vectors are referred to herein as "expression vectors".

[0153] The terms "host cell", "host cell line", and "host cell culture" are used interchangeably and refer to a cell into which exogenous nucleic acid has been introduced, including progeny of such cells.

[0154] In the present disclosure, "binding", "is bound", or "specifically binds", and similar terms, when referring to a protein and its ligand or an antibody and its antigen target, for example, or other binding pairs, mean that the binding affinity between members of the binding pair is strong enough that the interaction cannot be due to random molecular association (i.e., "non-specific binding"). Such binding typically requires a dissociation constant (K D ) of 1 μM or less and often can be accompanied by a K D of 100 nM or less.

[0155] "Affinity" refers to the overall strength of the non-covalent interaction 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 is generally the dissociation constant (K D) can be represented by. The affinity of an antibody for an antigen can be measured by common methods known in the art, such as surface plasmon resonance (SPR).

[0156] As used herein, the term "agonist" refers to a substance, such as an antibody, that causes an increase in at least one activity or function of the molecule to which it binds, or otherwise activates or helps to activate the molecule. The term "antagonist" or "inhibitor" as used herein refers to a substance, such as an antibody, that causes a decrease in at least one activity or function of the molecule to which it binds, or otherwise blocks or inhibits at least one activity or function of the molecule.

[0157] The term "inhibit" or "inhibiting" more generally refers to a decrease or cessation of any event (e.g., protein ligand binding), or a decrease or cessation of any phenotypic characteristic, or a decrease or cessation of the occurrence, degree, or likelihood of that characteristic. "Reduce" or "inhibit" means to decrease, lower, or stop an activity, function, and / or amount as compared to a reference. The inhibition or reduction need not be complete. For example, in some embodiments, "reduce" or "inhibit" means the ability to cause an overall decrease of 20% or greater. In another embodiment, "reduce" or "inhibit" means the ability to cause an overall decrease of 50% or greater. In yet another embodiment, "reduce" or "inhibit" means the ability to cause an overall decrease of 75%, 85%, 90%, 95%, or greater.

[0158] "Treatment", as used herein, encompasses any administration or application of a therapeutic agent for a disease in a human, and includes inhibiting the progression of a disease, or a disease or one or more disease symptoms, inhibiting or delaying a disease or its progression or one or more of its symptoms, halting its occurrence, partially or completely alleviating a disease or one or more of its symptoms, or preventing recurrence of one or more symptoms of a disease.

[0159] The terms "subject" and "patient" are used interchangeably herein and refer to a human unless otherwise clearly indicated (i.e., a mouse subject or the like).

[0160] "Autoimmune disease", or "autoimmune disorder", as used herein, encompasses diseases characterized by the subject'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. The present disclosure provides many non-limiting examples of autoimmune diseases throughout. 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.

[0161] The term "effective amount" or "therapeutically effective amount" refers to an amount of a drug that is effective for treating a disease or disorder in a subject, e.g., effective for partially or completely alleviating one or more symptoms. In some embodiments, an effective amount refers to an amount effective at the dosage and for the period required to achieve a desired therapeutic or prophylactic result.

[0162] "Biological sample", as used herein, refers to a sample taken from a subject 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. The biological sample may be taken directly from the subject or may be chemically or physically modified first in some way before use, for example, to assist in the analysis of the sample.

[0163] II. Exemplary anti-PAD4 antibodies Provided herein are antibodies that specifically bind to protein arginine deiminase 4 (PAD4). In some embodiments, the antibody inhibits the activity of PAD4, such as the citrullination of arginine.

[0164] Clone 13 and related antibodies The present disclosure relates to a group of antibodies based on a mouse anti-human antibody, such as that referred to as "Clone 13" and further described, for example, in Example 1 below. For example, Clone 13 was prepared in its original mouse anti-human form and then humanized to generate a series of antibodies designated hz13-1 to hz13-12, among which hz13-5 and hz13-12 were further modified at position 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 and paratope mapping to identify portions of the Clone 13 variable region and its humanized mutants that directly contact PAD4 provided additional structural information regarding portions of the Clone 13 variable region and its related humanized antibodies that direct 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 subsection and Example 14).

[0165] For example, in some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 9.

[0166] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 62, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 7, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 9.

[0167] In some embodiments, the present disclosure provides an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) having the amino acid sequence of SEQ ID NO: 4 or 62, and an HCDR3 having the amino acid sequence of SEQ ID NO: 6; and the light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) having the amino acid sequence of SEQ ID NO: 7. In particular, from the paratope mapping and structural analysis of the antibody comprising the above set of heavy and light chain CDRs, it was found that HCDR1, HCDR3, and LCDR1 contact PAD4 (see FIGS. 8A-8B). Thus, in some embodiments, the antibodies herein comprise the above set of these three CDRs. In some embodiments, the antibody comprises a VH comprising an HCDR1 having the amino acid sequence of SEQ ID NO: 4 and an HCDR3 having the amino acid sequence of SEQ ID NO: 6; and a VL comprising an LCDR1 having the amino acid sequence of SEQ ID NO: 7. In other embodiments, the antibody comprises a VH comprising an HCDR1 having the amino acid sequence of SEQ ID NO: 62 and an HCDR3 having the amino acid sequence of SEQ ID NO: 6; and a VL comprising an LCDR1 having the amino acid sequence of SEQ ID NO: 7.

[0168] In some embodiments, the antibody comprises a heavy chain variable region (VH) having the amino acid sequences of SEQ ID NO: 221 or 225 and SEQ ID NO: 222; and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO: 223. Each of these paratope regions was identified as contacting PAD4 as described in the Examples and FIGS. 8A-8B. In some embodiments, the antibody further comprises a light chain constant region having the amino acid sequence of SEQ ID NO: 224.

[0169] In some embodiments, the antibody comprises a VH that includes glycine (Gly94) at Kabat position 94 (see Figure 1E). Thus, for example, the originally identified murine anti-human clone 13 antibody includes glycine at Kabat position 94 but does not include some human framework regions. Thus, in certain humanized antibodies, a back mutation is required to obtain glycine at Kabat position 94. Gly94 is an amino acid residue in a loop adjacent to VH CDR3 (i.e., the VH CDR3 loop). In some embodiments, Gly94 is related to the mobility and / or geometry of the VH CDR3 loop. In some embodiments, Gly94 can interact with VH CDR3. In some embodiments, Gly94 is related to the activity of the anti-PAD4 antibody. In some embodiments, mutation of Gly94 to a different amino acid, such as threonine, results in a decrease in the mobility of the VH CDR3 loop, a change in the geometry of the VH CDR3 loop, a decrease in the interaction between the amino acid at position 94 and the VH CDR3 loop, a decrease in the binding of the anti-PAD4 antibody to PAD4, a decrease in the activity of the anti-PAD4 antibody, an increase in the binding of the anti-PAD4 antibody to extracellular matrix (ECM) proteins, a change in the secondary structure of the anti-PAD4 antibody, a decrease in the stability of the anti-PAD4 antibody, or any combination thereof. Thus, in some embodiments, the VH of the antibody includes glycine at Kabat position 94 (position 98 of SEQ ID NO: 10) (see Figure 1E for a depiction of this residue position located immediately prior to HCDR3).

[0170] 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 any one of the amino acid sequences of SEQ ID NO: 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 any one of the amino acid sequences of SEQ ID NO: 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 any one of the amino acid sequences of SEQ ID NO: 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 any one of the amino acid sequences of SEQ ID NO: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70. In some embodiments, the antibody comprises a VH comprising any one of the amino acid sequences of SEQ ID NO: 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 any one of the amino acid sequences 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 an 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, and a VL comprising an 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. In some such examples, the antibody VH and VL further comprise (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4 or 62, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, and an LCDR1 comprising the amino acid sequence of SEQ ID NO: 7; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4 or 62, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 7, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 8, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 9; or (c) further comprising 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, and further optionally comprising a light chain constant region comprising the amino acid sequence of SEQ ID NO: 224. Thus, in such examples, the mutations in VH and VL compared to the above sequence identifiers are located in regions outside of these specific CDR or paratope sequences. In some embodiments, the antibody comprises a VH comprising an 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 an 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.

[0171] In a further embodiment, the antibody comprises a VH comprising any one of the amino acid sequences of SEQ ID NO: 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, or 68; and a VL comprising any one of the amino acid sequences of SEQ ID NO: 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, or 70. Thus, for example, the following exemplary antibodies are within the scope of the present disclosure: - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), the isolated antibody comprising 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.

[0172] In some examples, in any of the above antibodies, the antibody binds to an epitope on PAD4 comprising SEQ ID NO: 217 and SEQ ID NO: 218.

[0173] In some embodiments of the present specification, the antibody is an IgA, IgG, or IgM antibody. In some examples, the antibody is an IgG antibody, such as a human IgG1, IgG2, IgG3, or IgG4 antibody, or a mouse IgG1 or IgG2 antibody. In some examples, 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 examples, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In still other examples, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some examples, the antibody is an antibody fragment, such as an Fv, single-chain Fv (scFv), Fab, Fab’, or (Fab’)2.

[0174] In some embodiments, the antibody is a bispecific or multispecific antibody or is conjugated to at least one other molecule by covalent or non-covalent bonding. In some embodiments, the antibody is conjugated to at least one other molecule by covalent or non-covalent bonding, and the at least one other molecule comprises a detection label and / or a drug.

[0175] In some embodiments, the antibody comprises a human IgG1 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 any one of the amino acid sequences 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 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. In some embodiments, the antibody comprises both a human IgG1 heavy chain constant region that is at least 90%, at least 95%, at least 97%, or at least 99% identical to any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and 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. In some examples, 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some examples, 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. In some examples, the antibody comprises both 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and 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.In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising any one amino acid sequence 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 any one amino acid sequence 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.

[0176] Clone 20 and related antibodies The present disclosure further relates to a second mouse anti-human antibody, clone 20, and its humanized mutants hz20-1 to hz20-14 described in the following examples. Thus, in some embodiments, the present disclosure provides an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 72, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 73, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 74; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 75, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 76, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 77.

[0177] 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 any one of the amino acid sequences of SEQ ID NO: 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 any one of the amino acid sequences of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136. In some embodiments, the antibody comprises a VH that comprises any one of the amino acid sequences of SEQ ID NO: 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 the antibody comprises a VL that comprises any one of the amino acid sequences of SEQ ID NO: 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. In some such examples, the antibody VH further comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 72, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 73, or an HCDR3 comprising the amino acid sequence of SEQ ID NO: 74. In some examples, the antibody VL further comprises an LCDR1 comprising the amino acid sequence of SEQ ID NO: 75, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 76, or an LCDR3 comprising the amino acid sequence of SEQ ID NO: 77. In some embodiments, the antibody comprises a VH that comprises any one of the amino acid sequences of SEQ ID NO: 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 comprises any one of the amino acid sequences of SEQ ID NO: 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, or 136.

[0178] In yet further 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, for example, the following exemplary antibodies are within the scope of the present disclosure: - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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; - An isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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.

[0179] In some examples, in any of the above antibodies, the antibody binds to an epitope on PAD4 comprising SEQ ID NO: 219 and SEQ ID NO: 220.

[0180] In some embodiments of the present specification, the antibody is an IgA, IgG, or IgM antibody. In some examples, the antibody is an IgG antibody, such as a human IgG1, IgG2, IgG3, or IgG4 antibody, or a mouse IgG1 or IgG2 antibody. In some examples, the antibody comprises a wild-type, human IgG1, IgG2, or IgG4 heavy chain constant region. In certain embodiments, the antibody is an antibody of the human IgG1 isotype. In certain embodiments, the antibody is an antibody of the human IgG1 isotype having P329G, L234A, and L235A (LALAPG; EU numbering) mutations to reduce the Fc region effector function. In other embodiments, the antibody is an antibody of the human IgG2 isotype. In certain embodiments, the antibody is an antibody of the IgG4 isotype having an S228P mutation (EU numbering) in the hinge region to improve the stability of the IgG4 antibody. In some such embodiments, the antibody (e.g., a non-humanized antibody) may have a non-human IgG constant region, for example, a mouse IgG2a antibody, such as a mouse IgG2a LALAPG antibody. In some embodiments, the antibody comprises a full-length heavy chain and / or a full-length light chain. In other examples, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In still other examples, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some examples, the antibody is an antibody fragment, such as an Fv, single-chain Fv (scFv), Fab, Fab’, or (Fab’)2.

[0181] In some embodiments, the antibody is a bispecific or multispecific antibody, or is conjugated to at least one other molecule by a covalent or non-covalent bond. In some embodiments, the antibody is conjugated to at least one other molecule by a covalent or non-covalent bond, and the at least one other molecule comprises a detection label and / or a drug.

[0182] In some embodiments, the antibody comprises a human IgG1 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 any one of the amino acid sequences 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 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. In some embodiments, the antibody comprises both a human IgG1 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and 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. In some examples, 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some examples, 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. In some examples, the antibody comprises both 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and 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.In some embodiments, the antibody comprises a human IgG1 heavy chain constant region comprising any one amino acid sequence 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 the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antibody comprises both a human IgG1 heavy chain constant region comprising any one amino acid sequence of SEQ ID NOs: 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.

[0183] pH-Dependent Clone 13-Related Antibody In some embodiments, the present disclosure relates to one or more antibody variants of parental antibody clone 13 or hz13-5, wherein specific residues are modified as described in Example 14 below. In certain instances, although not in all instances, these modifications have been shown to affect the pH-dependence of PAD4 binding as described in Example 14.

[0184] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a VH comprising an HCDR1 comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 138, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 138, and an HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 138. In some of the above embodiments, the antibody further comprises a VL comprising an LCDR1 comprising the amino acid sequence at residues 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence at residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence at residues 93-101 of SEQ ID NO: 170. In other embodiments of the above, the antibody comprises a VL comprising an LCDR1 comprising the amino acid sequence at residues 24-38 of SEQ ID NO: 172, an LCDR2 comprising the amino acid sequence at residues 54-60 of SEQ ID NO: 172, and an LCDR3 comprising the amino acid sequence at residues 93-101 of SEQ ID NO: 172.

[0185] In some embodiments, the antibody is - a VH comprising an HCDR1 comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 138, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 138, and an HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 138, and a VL comprising an LCDR1 comprising the amino acid sequence at residues 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence at residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence at residues 93-101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 140, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 140, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 140, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 142, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 142, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 142, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 144, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 144, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 144, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 146, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 146, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 146, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 148, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 148, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 148, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 150, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 150, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 150, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 152, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 152, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 152, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 154, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 154, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 154, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 156, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 156, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 156, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 158, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 158, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 158, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 160, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 160, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 160, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 162, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 162, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 162, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 164, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 164, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 164, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 166, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 166, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 166, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 168, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 168, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 168, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 170, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 170 comprising.

[0186] In some embodiments, the antibody is - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 138, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 138, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 138, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 140, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 140, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 140, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 142, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 142, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 142, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 144, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 144, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 144, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 146, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 146, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 146, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 148, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 148, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 148, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 150, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 150, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 150, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 152, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 152, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 152, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 154, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 154, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 154, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 156, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 156, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 156, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 158, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 158, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 158, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 160, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 160, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 160, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 162, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 162, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 162, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 164, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 164, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 164, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 166, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 166, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 166, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172; - A VH comprising an HCDR1 containing the amino acid sequence at positions 26 to 35 of SEQ ID NO: 168, an HCDR2 containing the amino acid sequence at positions 50 to 66 of SEQ ID NO: 168, and an HCDR3 containing the amino acid sequence at positions 99 to 108 of SEQ ID NO: 168, and a VL comprising an LCDR1 containing the amino acid sequence of residues 24 to 38 of SEQ ID NO: 172, an LCDR2 containing the amino acid sequence of residues 54 to 60 of SEQ ID NO: 172, and an LCDR3 containing the amino acid sequence of residues 93 to 101 of SEQ ID NO: 172 is included.

[0187] 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 that is 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 provided above.

[0188] 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 also comprises 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 also comprises a VL comprising the amino acid sequence of SEQ ID NO: 172.

[0189] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 138, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 138, and an HCDR3 comprising positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 138 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0190] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0191] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0192] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4) and comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 140, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 140, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 140 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 140 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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 that comprises 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 that comprises 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. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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.

[0193] In some embodiments, the antibody comprises a VH that comprises the amino acid sequence of SEQ ID NO: 140. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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: 140; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both a VH that comprises the amino acid sequence of SEQ ID NO: 140, and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0194] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0195] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 142, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 142, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 142 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 142 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0196] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0197] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0198] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 144, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 144, and an HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 144; and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 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 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 144 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions, and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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 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: 172 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 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.

[0199] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0200] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0201] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of positions 26-35 of SEQ ID NO: 146, an HCDR2 comprising the amino acid sequence of positions 50-66 of SEQ ID NO: 146, and an HCDR3 comprising the amino acid sequence of positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 146 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 146 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0202] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0203] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0204] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) containing a heavy chain complementarity determining region 1 (HCDR1) with the amino acid sequence at positions 26-35 of SEQ ID NO: 148, an HCDR2 with the amino acid sequence at positions 50-66 of SEQ ID NO: 148, and an HCDR3 with the amino acid sequence at positions 99-108 of SEQ ID NO: 148; and a light chain variable region (VL) containing a light chain complementarity determining region 1 (LCDR1) with the amino acid sequence of residues 24-38 of SEQ ID NO: 170, an LCDR2 with the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 with the amino acid sequence of residues 93-101 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 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: 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 containing the amino acid sequence of SEQ ID NO: 148 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL containing the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH containing the amino acid sequence of SEQ ID NO: 148 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL containing the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 that comprises 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 that comprises 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. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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.

[0205] In some embodiments, the antibody comprises a VH that comprises the amino acid sequence of SEQ ID NO: 148. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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: 148; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both a VH that comprises the amino acid sequence of SEQ ID NO: 148; and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0206] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0207] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 150, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 150, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 150 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0208] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0209] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0210] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising 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, an HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 152, and an 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, an LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an 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 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: 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 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 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: 170 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 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: 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 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: 172 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 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.

[0211] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0212] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0213] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) including a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 154, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 154, and an HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 154; and a light chain variable region (VL) including a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 154 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 154 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0214] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0215] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0216] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 156, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 156, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 156 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions, and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0217] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0218] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0219] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 158, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 158, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 158 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 158 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0220] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0221] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0222] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 160, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 160, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 160 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 160 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions, and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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 that comprises 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 that comprises 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. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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.

[0223] In some embodiments, the antibody comprises a VH that comprises the amino acid sequence of SEQ ID NO: 160. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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: 160; and a VL that comprises the amino acid sequence of SEQ ID NO: 170. In some embodiments, the antibody comprises both a VH that comprises the amino acid sequence of SEQ ID NO: 160, and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0224] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0225] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4) and comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 162, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 162, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 162 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 162 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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 that comprises 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 that comprises 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. In some embodiments, the antibody comprises both a VH that comprises 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 that comprises 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.

[0226] In some embodiments, the antibody comprises a VH that comprises the amino acid sequence of SEQ ID NO: 162. In some embodiments, the antibody comprises a VH that comprises 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 that comprises 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 that comprises the amino acid sequence of SEQ ID NO: 162, and a VL that comprises the amino acid sequence of SEQ ID NO: 170.

[0227] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0228] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4) and 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, an HCDR2 comprising the amino acid sequence of positions 50 to 66 of SEQ ID NO: 164, and an 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, an LCDR2 comprising the amino acid sequence of residues 54 to 60 of SEQ ID NO: 170, and an 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 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: 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 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 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: 170 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 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 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: 172 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 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.

[0229] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0230] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0231] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) including a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 166, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 166, and an HCDR3 comprising the amino acid sequence at positions 99-108 of SEQ ID NO: 166; and a light chain variable region (VL) including a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 166 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions, and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0232] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0233] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0234] In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to protein arginine deiminase 4 (PAD4), comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence at positions 26-35 of SEQ ID NO: 168, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 168, and an HCDR3 comprising the amino acid sequence at positions 99-108 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 24-38 of SEQ ID NO: 170, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 170, and an LCDR3 comprising the amino acid sequence of residues 93-101 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 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: 170. In some embodiments, the antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 168 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions. In some embodiments, the antibody comprises both a VH comprising the amino acid sequence of SEQ ID NO: 168 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 amino acid substitutions and a VL comprising the amino acid sequence of SEQ ID NO: 170 modified by 1-10 amino acid substitutions, 1-5 amino acid substitutions, or 1-3 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: 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 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: 172 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 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.

[0235] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 170.

[0236] 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 comprising 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 comprising the amino acid sequence of SEQ ID NO: 172.

[0237] In some examples, the antibody is an IgG antibody, such as a human IgG1, IgG2, IgG3, or IgG4 antibody, or a mouse IgG1 or IgG2 antibody. In some examples, 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 examples, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In still other examples, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some examples, the antibody is an antibody fragment, such as an Fv, single-chain Fv (scFv), Fab, Fab’, or (Fab’)2.

[0238]

[0239] In some embodiments, the antibody is a bispecific or multispecific antibody, or is conjugated to at least one other molecule by covalent or non-covalent bonding. In some embodiments, the antibody is conjugated to at least one other molecule by covalent or non-covalent bonding, and the at least one other molecule comprises a detection label and / or a drug.In some embodiments, the antibody comprises a human IgG1 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 any one of the amino acid sequences 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 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. In some embodiments, the antibody comprises both a human IgG1 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and 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. In some examples, 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192. In some examples, 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. In some examples, the antibody comprises both 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 any one of the amino acid sequences of SEQ ID NO: 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192; and a light chain constant region comprising an amino acid sequence modified by 1 - 10 amino acid substitutions, 1 - 5 amino acid substitutions, or 1 - 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 any one amino acid sequence 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 any one amino acid sequence 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.

[0240] Mouse antibody In some embodiments, the present disclosure relates to an isolated antibody that specifically binds to mouse protein arginine deiminase 4 (PAD4), comprising a VH comprising an HCDR1 comprising the amino acid sequence at positions 31-35 of SEQ ID NO: 208, an HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 208, and an HCDR3 comprising the amino acid sequence at positions 99-107 of SEQ ID NO: 208; and a light chain constant region (VL) comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 210, an LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 210, and an LCDR3 comprising the amino acid sequence of residues 93-101 of SEQ ID NO: 210.

[0241] 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. 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 that comprises the 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 that comprises any one of the amino acid sequences of SEQ ID NO: 210 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 that comprises the 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; and a VL that comprises any one of the amino acid sequences of SEQ ID NO: 210 modified by 1 to 10 amino acid substitutions, 1 to 5 amino acid substitutions, or 1 to 3 amino acid substitutions. In some such examples, the antibody further comprises HCDR1 comprising the amino acid sequence at positions 31-35 of SEQ ID NO: 208, HCDR2 comprising the amino acid sequence at positions 50-66 of SEQ ID NO: 208, and HCDR3 comprising the amino acid sequence at positions 99-107 of SEQ ID NO: 208, LCDR1 comprising the amino acid sequence of residues 24-38 of SEQ ID NO: 210, LCDR2 comprising the amino acid sequence of residues 54-60 of SEQ ID NO: 210, and LCDR3 comprising the amino acid sequence of residues 93-101 of SEQ ID NO: 210.

[0242] 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.

[0243] In some embodiments herein, the antibody is an IgA, IgG, or IgM antibody. In some examples, the antibody is a mouse IgG1 or IgG2 antibody. In some embodiments, the antibody comprises a full-length heavy chain and / or a full-length light chain. In other examples, the antibody lacks a C-terminal lysine at the end of the heavy chain constant region. In still other examples, the antibody lacks a C-terminal glycine-lysine at the end of the heavy chain constant region. In some examples, the antibody is an antibody fragment, such as an Fv, single-chain Fv (scFv), Fab, Fab’, or (Fab’)2.

[0244] In some embodiments, the antibody is a bispecific or multispecific antibody, or is conjugated to at least one other molecule by a covalent or non-covalent bond. In some embodiments, the antibody is conjugated to at least one other molecule by a covalent or non-covalent bond, and the at least one other molecule comprises a detection label and / or a drug.

[0245] In some embodiments, the antibody comprises a HC comprising the amino acid sequence of SEQ ID NO: 212. In some embodiments, the antibody comprises an 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 an LC comprising the amino acid sequence of SEQ ID NO: 214.

[0246] III. Exemplary Antibody Variants, Fragments, and Constant Regions In many embodiments, an antibody that specifically binds to PAD4 may further incorporate any one or combination of the features described in the following sections.

[0247] A. Antibody Fragments 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, Pluckthuen, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315(1994); similarly WO93 / 16185; and U.S. Pat. Nos. 5,571,894 and 5,877,458. For a discussion of Fab and F(ab”)2 fragments that contain salvage receptor binding epitope residues and have increased in vivo half-lives, see U.S. Pat. No. 5,869,046.

[0248] A diabody is an antibody fragment that has two antigen-binding sites that can be bivalent or bispecific. See, for example, European Patent No. 404,097; WO1993 / 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).

[0249] A single domain antibody is an antibody fragment that comprises all or part of the heavy chain variable domain or all or part 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, for example, U.S. Pat. No. 6,248,516).

[0250] Antibody fragments can be made by a variety of techniques including, but not limited to, proteolytic digestion of intact antibodies as well as production by recombinant host cells [e.g., E. coli or phage] described herein.

[0251] B. Bispecific or multispecific antibodies In certain embodiments, the antibodies provided herein are multispecific antibodies, e.g., bispecific antibodies. Multispecific antibodies are monoclonal antibodies having binding specificities 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, the bispecific antibody can bind to two different epitopes of TREM2. Bispecific antibodies can also be used to localize drugs such as cytotoxic agents or to localize detection labels to cells expressing TREM2. In some embodiments, the multispecific antibody (e.g., bispecific antibody) comprises a first variable domain comprising a CDR or variable region described herein. Bispecific antibodies can be prepared as full-length antibodies or antibody fragments.

[0252] Techniques for making bispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs having different specificities (see Milstein and Cuello, Nature 305:537 (1983)), WO93 / 08829, and Traunecker et al., EMBO J. 10:3655 (1991)), and "knob-into-hole" engineering (see, e.g., U.S. Patent No. 5,731,168). Bispecific antibodies include engineering of electrostatic steering effects for making antibody Fc-heterodimer molecules (WO2009 / 089004A1); cross-linking of two or more antibodies or fragments (see, e.g., U.S. Patent No. 4,676,980, and Brennan et al., Science, 229:81 (1985)); use of leucine zippers for producing bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 18(5):1547-1553 (1992)); use of the "diabod" technique for making bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and use of single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and can also be made by, for example, the preparation of trispecific antibodies described in Tutt et al. J. Immunol. 147:60 (1991).

[0253] Engineered antibodies having three or more functional antigen-binding sites, including "octopus antibodies", are also included herein (see, e.g., U.S. Patent Application Publication No. 2006 / 0025576).

[0254] C. Chimeric and Humanized Antibodies In certain embodiments, the antibodies provided herein are chimeric antibodies. Certain chimeric antibodies are described, for example, in U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In one example, 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 a further example, a chimeric antibody is a "class switch" antibody in which the class or subclass has changed from that of the parental antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0255] In certain embodiments, the chimeric antibody is a humanized antibody. Typically, non-human antibodies are humanized to reduce their immunogenicity in humans while retaining the specificity and affinity of the parental non-human antibody. Generally, a humanized antibody comprises one or more variable domains in which the HVRs, e.g., CDRs (or a portion thereof), are derived from a non-human antibody and the FRs (or a portion thereof) are derived from a human antibody sequence. A humanized antibody may also comprise 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., the antibody from which the HVR residues are derived) to, for example, restore or improve the specificity or affinity of the antibody.

[0256] Humanized antibodies and methods for making them are reviewed, for example, in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and are 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 "guide selection" approach to FR shuffling].

[0257] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best fit" method (e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from consensus sequences of human antibodies of certain subgroups of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatic 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 of FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0258] In some embodiments, the humanized antibody can include a human IgG1, IgG2, IgG3, or IgG4 heavy chain constant region.

[0259] D. Glycosylation and PEGylation Variants In certain embodiments, the glycosylation of the antibody is modified. For example, deglycosylated antibodies can be made (i.e., the antibody lacks glycosylation). The glycosylation can be altered, for example, to increase the affinity of the antibody for an antigen. Such carbohydrate modifications can be achieved, for example, by altering one or more sites of glycosylation within the antibody sequence. For example, removal of one or more variable region framework glycosylation sites can be effected and one or more amino acid substitutions can be made to remove glycosylation at that site. Such deglycosylation can increase the affinity of the antibody for an antigen. Such an approach is described in more detail in U.S. Pat. Nos. 5,714,350 and 6,350,861 by Co et al.

[0260] Glycosylation of the constant region at N297 can be prevented by mutating the N297 residue to another residue, e.g., N297A, and / or by mutating an adjacent amino acid, e.g., 298, thereby reducing glycosylation at N297.

[0261] In addition or alternatively, antibodies having an altered type of glycosylation can be produced, such as afucosylated antibodies having a reduced amount of fucosyl residues, or antibodies having an increased bisected GlcNAc structure. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of the antibody. Such sugar chain modifications can be achieved, for example, by expressing the antibody in a host cell by an altered glycosylation mechanism. Cells having an altered glycosylation mechanism are described in the art. The recombinant antibodies described herein can be used as host cells for expressing the antibody and thereby producing an antibody having altered glycosylation. For example, European Patent No. 1,176,195 by Hanai et al. describes a cell line in which the FUT8 gene encoding fucosyltransferase is functionally disrupted such that the antibody expressed in such cell lines exhibits afucosylation. PCT Application WO03 / 035835 by Presta describes a mutant CHO cell line, Led3 cells, in which the ability to attach fucose to the Asn(297)-linked sugar chain is reduced, resulting in afucosylation of the antibody expressed in the host cell as well. (See also Shields, R.L. et al. (2002) J. Biol. Chem. 277:26733-26740). PCT Application WO99 / 54342 by Umana et al. describes a cell line engineered to express a glycoprotein-modifying glycosyltransferase (e.g., beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) such that the antibody expressed in the engineered cell line exhibits an increase in bisected GlcNAc structure resulting in an increase in the ADCC activity of the antibody. (See also Umana et al. (1999) Nat. Biotech. 17:176-180).

[0262] Another modification of the antibodies described herein is pegylation. The antibody can be pegylated, for example, to increase the biological (e.g., in serum) half-life of the antibody. To pegylate the antibody, the antibody or fragment thereof is reacted with polyethylene glycol (PEG), e.g., a reactive ester or aldehyde derivative of PEG, under conditions such that one or more PEG groups become attached to the antibody or antibody fragment. In some embodiments, pegylation is effected via an acylation or alkylation reaction with a reactive PEG molecule (or similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any type of PEG that has been used to derivatize other proteins, such as mono(CI-CIO)alkoxy- or aryloxy-polyethylene glycol, or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is a deglycosylated antibody. Methods for pegylating proteins are known in the art and can be applied to the antibodies described herein. See, for example, European Patent No. 0 154 316 by Nishimura et al. and European Patent No. 0 401 384 by Ishikawa et al.

[0263] E. Constant region In some embodiments, the antibodies described herein include one or more human constant regions. In some embodiments, the human heavy chain constant region is a region of an isotype selected from IgA, IgG, and IgD. In some embodiments, the human light chain constant region is a region of an isotype selected from κ and λ. In some embodiments, the antibodies described herein include a human IgG constant region, e.g., IgG1, IgG2, IgG3, or IgG4. In some embodiments, the antibodies described herein include a human IgG4 heavy chain constant region. In some such embodiments, the antibodies described herein include the S241P mutation in the human IgG4 constant region. In some embodiments, the antibodies described herein include a human IgG4 constant region and a human κ light chain.

[0264] The selection of the heavy chain constant region can determine whether the antibody has effector function in vivo. Such effector functions, in some embodiments, include antibody-dependent cell-mediated cytotoxicity (ADCC) and / or complement-dependent cytotoxicity (CDC), which can result in the killing of the cells to which the antibody binds. In some treatment methods, including some methods of treating certain cancers, the killing of cells may be desirable, for example, when the antibody binds to cells that support tumor maintenance or growth. Exemplary cells that can support tumor maintenance or growth include, but are not limited to, the tumor cells themselves, cells that assist in the recruitment of the vasculature to the tumor, and the details tail that provides ligands, growth factors, or counter-receptors that support or promote tumor growth or tumor survival. In some embodiments, when effector function is desired, an antibody comprising a human IgG1 heavy chain or a human IgG3 heavy chain is selected.

[0265] In certain embodiments, the antibodies provided herein are modified to increase or decrease the degree to which the antibody is glycosylated. The addition or deletion of glycosylation sites to the antibody can be readily achieved by altering the amino acid sequence such that one or more glycosylation sites are created or removed.

[0266] If the antibody contains an Fc region, the sugar chains bound thereto may be modified. Natural antibodies produced by mammals typically contain branched, biantennary oligosaccharides that are commonly bound by N-linkage to Asn297 in the CH2 domain of the Fc region. See, for example, Wright et al. TIBTECH 15:26-32 (1997). The oligosaccharides can include various sugar chains such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose bound to GlcNAc in the "stem" of the biantennary oligosaccharide structure. In some embodiments, modification of the oligosaccharides in the antibodies of the present invention can be performed to produce antibodies having certain improved properties. For example, in some embodiments, the antibody may be defucosylated by mutating residues such as Asn297 that are normally glycosylated by fucose-containing glycosylation, or through other means. In some embodiments, the antibodies herein may include a defucosylated human IgG1 constant region.

[0267] For example, there is further provided an antibody having a biantennary oligosaccharide in which the biantennary oligosaccharide bound to the Fc region of the antibody is bisected by GlcNAc. Such antibodies may have reduced fucosylation and / or improved ADCC function. Examples of such antibodies are described, for example, in WO2003 / 011878 (Jean-Mairet et al.); U.S. Patent No. 6,602,684 (Umana et al.); and U.S. Patent Application Publication No. 2005 / 0123546 (Umana et al.). Antibodies in which at least one galactose residue in the oligosaccharide is bound to the Fc region are also provided. Such antibodies may have improved CDC function. Such antibodies are described, for example, in WO1997 / 30087 (Patel et al.); WO1998 / 58964 (Raju, S.); and WO1999 / 22764 (Raju, S.).

[0268] Antibodies having an amino-terminal leader extension are also provided. For example, one or more amino acid residues of the amino-terminal leader sequence are present at the amino terminus of one or more heavy or light chains of the antibody. Exemplary amino-terminal leader extensions include or consist of three amino acid residues present on one or both light chains of the antibody, VHS.

[0269] The in vivo or serum half-life of a human FcRn high-affinity binding polypeptide can be assayed, for example, in transgenic mice, humans, or non-human primates that are administered a polypeptide having a variant Fc region. See, similarly, for example, Petkova et al. International Immunology 18(12):1759-1769 (2006).

[0270] In some embodiments of the invention, the defucosylated antibody mediates ADCC more effectively than the fucose-containing parental antibody in the presence of human effector cells. In general, ADCC activity can be determined using the in vitro ADCC assays disclosed herein, although other assays or methods for determining ADCC activity, such as in animal models, are contemplated.

[0271] In certain embodiments, the Fc region is modified by replacing at least one amino acid residue with a different amino acid residue to alter 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) can be replaced with different amino acid residues such that the antibody has an altered affinity for an effector ligand but retains the antigen-binding ability of the parental antibody. The effector ligand with altered affinity can be, for example, an Fc receptor or the C1 component of complement. This approach is described in more detail in U.S. Pat. Nos. 5,624,821 and 5,648,260 by Winter et al.

[0272] In some examples, one or more amino acids selected from amino acid residues 329, 331, and 322 can be replaced with different amino acid residues such that the antibody has altered C1q binding and / or reduced or abolished complement-dependent cytotoxicity (CDC). This approach is described in more detail in U.S. Patent No. 6,194,551 by Idusogie et al.

[0273] In some examples, one or more amino acid residues within amino acid positions 231 and 239 are altered, thereby altering the ability of the antibody to fix complement. This approach is described in detail in PCT application WO94 / 29351 by Bodmer et al. In some examples, the Fc region is modified 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 numbering) to reduce antibody-dependent cell cytotoxicity (ADCC) and / or modify to reduce the affinity for Fcγ receptors. 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 / 268F7324T (EU numbering). Other Fc modifications that can be made to Fc are modifications to reduce or eliminate binding to FcγR 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), and the numbering follows 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), and the numbering follows the EU index. The Fc variant may include 236R / 328R. Other modifications for reducing FcγR and complement interaction include the substitutions 297A, 234A, 235A, 237A, 318A, 228P, 236E, 268Q, 309L, 330S, 331S, 220S, 226S, 229S, 238S, 233P, and 234V, and removal of glycosylation at position 297 by mutation or enzymatic means or by production in organisms such as bacteria that do not glycosylate the protein. 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 L234A, L235E, and G237A substitutions. IgG1fa.P238K (or IgG1.P238K) contains the P238K substitution. IgG1.1f includes the L234A, L235E, G237A, A330S, and P331S substitutions (all numbering according to the EU index).

[0274] Fc variants that enhance the affinity for the inhibitory receptor FcγRIIb can also be used. Such variants can provide, for example, Fc fusion proteins having immunomodulatory activities associated with FcγRIIb cells, including B cells and monocytes. In one embodiment, the Fc variant provides selectively enhanced affinity for FcγRIIb compared to one or more activating receptors. Modifications that alter the 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 that enhance FcγRllb affinity 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 enhancing the binding to FcγRIIb include 235Y / 267E, 236D / 267E, 239D / 268D, 239D / 267E, 267E / 268D, 267E / 268E, and 267E / 328F (all numbering is according to the EU index).

[0275] Other modifications to enhance FcγR and complement interactions 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 amino acid positions 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, 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 in the Fc region, and the numbering of residues in the Fc region is the EU index numbering as described in patent application WO00 / 42072.

[0276] The Fc region may contain amino acid residues that are not naturally present at additional and / or alternative positions known to those skilled in the art (see, for example, U.S. Patent Nos. 5,624,821; 6,277,375; 6,737,056; 6,194,551; 7,317,091; 8,101,720; PCT patent applications WO00 / 42072; WO01 / 58957; WO02 / 06919; WO04 / 016750; WO04 / 029207; WO04 / 035752; WO04 / 074455; WO04 / 099249; WO04 / 063351; WO05 / 070963; WO05 / 040217; WO05 / 092925 and WO06 / 020114).

[0277] The affinity and binding characteristics of the Fc region to the ligand can be determined by a variety of in vitro assay methods (biochemical or immunology-based methods) known in the art, including but not limited to equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA), or radioimmunoassay (RIA)), or kinetics (e.g., BIACORE analysis), and 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 can utilize labels on one or more of the components being tested and use a variety of detection methods including chromogenic, fluorescent, luminescent, or isotope labeling. A detailed description of binding affinity and kinetics can be found in Paul, W. E., ed., Fundamental immunology, 4th Ed., Lippincott-Raven, Philadelphia (1999), which focuses on antibody-immunogen interactions.

[0278] In certain embodiments, the antibody is modified to increase its biological half-life. Various approaches are possible. For example, this can be done by increasing the binding affinity of the Fc region for FcRn, for example, by mutating one or more of the following residues: 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 increase the biological half-life, the antibody can be modified within the CH1 or CL region to contain a salvage receptor binding epitope derived from two loops of the CH2 domain of the Fc region of IgG as described in U.S. Patents Nos. 5,869,046 and 6,121,022 by 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, 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, 2001, 276(9):6591-6604), 252F, 252T, 252Y, 252W, 254T, 256S, 256R, 256Q, 256E, 256D, 256T, 309P, 311S, 433R, 433S, 4331, 433P, 433Q, 434H, 434F, 434Y, 252Y / 254T / 256E, 433K / 434F / 436H, 308T / 309P / 311S (Dall’Acqua et al. Journal of Immunology, 2002, 169:5171-5180, Dal’Acqua et al., 2006, Journal of Biological Chemistry 281:23514-23524). Other modifications that modulate FcRn binding are described in Yeung et al., 2010, J Immunol, 182:7663-7671.

[0279] In certain embodiments, hybrid IgG isotypes having specific biological characteristics can be used. For example, IgG1 / IgG3 hybrid variants can be constructed by substituting the IgG1 positions in the CH2 and / or CH3 regions with amino acids from IgG3 at positions where the two isotypes differ. Thus, hybrid variant IgG antibodies can be constructed that include one or more substitutions, such as 274Q, 276K, 300F, 339T, 356E, 358M, 384S, 392N, 397M, 422I, 435R, and 436F. In some embodiments described herein, IgG1 / IgG2 hybrid variants can be constructed by substituting the IgG2 positions in the CH2 and / or CH3 regions with amino acids from IgG1 at positions where the two isotypes differ. Thus, hybrid variant IgG antibodies can be constructed that include one or more substitutions, such as one or more of the following amino acid substitutions: 233E, 234L, 235L, +236G (referring to the insertion of glycine at position 236), and 327A.

[0280] In addition, the binding sites on human IgG1 for FcγRI, FcγRII, FcγRIII and FcRn have been mapped and variants with improved binding have been described (see Shields, R.L. 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. In addition, the following combinatorial mutants have been shown to improve FcγRIII binding: T256A / S298A, S298A / E333A, S298A / K224A and S298A / E333A / K334A, which have been shown to exhibit enhanced FcγRIIIa activity and ADCC activity (Shields et al., 2001). Other IgG1 variants with strongly enhanced binding to FcγRIIIa have been identified, including variants with the S239D / I332E and S239D / I332E / A330L mutations, which showed the greatest increase in affinity for FcγRIIIa, a decrease in FcγRIIb binding, and strong cytotoxic activity in cynomolgus monkeys (Lazar et al., 2006). Introduction of three mutations into antibodies such as alemtuzumab (CD52-specific), trastuzumab (HER2 / neu-specific), rituximab (CD20-specific), and cetuximab (EGFR-specific) resulted in a large enhancement of ADCC activity in vitro, and the S239D / I332E variant showed enhanced ability to deplete B cells in monkeys (Lazar et al., 2006). In addition, IgG1 mutants containing the L235V, F243L, R292P, Y300L and P396L mutations have been identified that showed enhanced binding to FcγRIIIa and simultaneous enhancement of ADCC activity in transgenic mice expressing human FcγRIIIa in B cell malignancies and breast cancer models (Stavenhagen et al., 2007; Nordstrom et al., 2011). Other Fc mutants that can be used are.It includes S298A / E333A / L334A, S239D / I332E, S239D / I332E / A330L, L235V / F243L / R292P / Y300L / P396L, and M428L / N434S.

[0281] In certain embodiments, an Fc with reduced binding to FcγR is selected. Exemplary Fcs, e.g., an IgG1 Fc with reduced FcγR binding, include the following three amino acid substitutions: L234A, L235E, and G237A.

[0282] In certain embodiments, an Fc with reduced complement fixation is selected. Exemplary Fcs, e.g., an IgG1 Fc with reduced complement fixation, have the following two amino acid substitutions: A330S and P331S.

[0283] In certain embodiments, an Fc that is essentially effector function - free is selected, i.e., the Fc has reduced binding to FcγR and reduced complement fixation. Exemplary Fcs that are effector - function - free, e.g., an IgG1 Fc, include the following five mutations: L234A, L235E, G237A, A330S, and P331S.

[0284] When using the IgG4 constant domain, it may include the substitution S228P that mimics the hinge sequence in IgG1 and thereby stabilizes the IgG4 molecule. Fc modifications described in WO2017 / 087678 or WO2016081746 may also be used.

[0285] In certain embodiments, the glycosylation of the antibody is modified. For example, deglycosylated antibodies can be produced (i.e., the antibody lacks glycosylation). The glycosylation can be altered, for example, to increase the affinity of the antibody for the antigen. Such glycan modifications increase the affinity of the antibody for the antigen. Such glycan 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 that result in the removal of one or more variable region framework glycosylation sites, thereby removing the glycosylation at that site. Such deglycosylation can increase the affinity of the antibody for the antigen. Such an approach is described in more detail in U.S. Pat. Nos. 5,714,350 and 6,350,861 by Co et al.

[0286] Glycosylation of the constant region at N297 can be prevented by mutating the N297 residue to another residue, e.g., N297A, and / or mutating an adjacent amino acid, e.g., 298, thereby reducing the glycosylation at N297.

[0287] In addition or alternatively, antibodies having an altered type of glycosylation can be produced. For example, hypofucosylated antibodies having a reduced amount of fucosyl residues or antibodies having an increased bisected GlcNac structure can be generated. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of the antibody. Such glycan modifications can be achieved, for example, by expressing the antibody in a host cell having an altered glycosylation machinery. Cells having an altered glycosylation machinery are described in the art and can be used as host cells for expressing the recombinant antibodies described herein and thereby producing antibodies having the altered glycosylation. For example, European Patent No. 1,176,195 by Hanai et al. describes cell lines in which the FUT8 gene encoding fucosyltransferase is functionally disrupted such that the antibodies expressed in such cell lines exhibit hypofucosylation. PCT Application WO03 / 035835 by Presta describes a mutant CHO cell line, Led3 cells, which have a reduced ability to attach fucose to the Asn(297)-linked glycans and which likewise result in hypofucosylation of the antibodies expressed in that host cell (see also Shields, R.L. et al. (2002) J. Biol. Chem. 277:26733-26740). PCT Application WO99 / 54342 by Umana et al. describes cells engineered to express a glycoprotein-modifying glycosyltransferase (e.g., beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII)) such that the antibodies expressed in the engineered cell lines exhibit an increase in the bisected GlcNac structure, which results in an increase in the ADCC activity of the antibody (see also Umana et al. (1999) Nat. Biotech. 17:176-180).

[0288] Another modification of the antibodies described herein is pegylation. The antibody can be pegylated, for example, to increase the biological (e.g., serum) half-life of the antibody. To pegylate the antibody, the antibody or a fragment thereof is reacted with polyethylene glycol (PEG), e.g., a reactive ester or aldehyde derivative of PEG, under conditions such that one or more PEG groups become attached to the antibody or antibody fragment. In some embodiments, pegylation is effected via an acylation or alkylation reaction with a reactive PEG molecule (or similar reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any form of PEG that has been used to derivatize other proteins, such as mono(CI-CIO)alkoxy- or aryloxy-polyethylene glycol, or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is a deglycosylated antibody. Methods for pegylating proteins are known in the art and can be applied to the antibodies described herein. See, for example, European Patent No. 0 154 316 by Nishimura et al. and European Patent 0 401 384 by Ishikawa et al.

[0289] IV. Exemplary Properties of Specific Anti-PAD4 Antibodies In many embodiments, an antibody that specifically binds to PAD4 can include any of the following properties, alone or in combination.

[0290] In some examples, the binding of an antibody to a ligand, such as PAD4, may be determined by surface plasmon resonance (SPR). In some embodiments, the antibody has a K D for specific binding to PAD4 of less than 5 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, from 0.01 nM to 5 nM, from 0.01 nM to 1 nM, from 0.05 nM to 1 nM, from 0.1 nM to 1 nM, from 0.1 nM to 0.5 nM, from 0.05 nM to 0.5 nM, from 0.05 nM to 0.1 nM, or from 0.5 nM to 1 nM in an SPR assay. For example, such a K Dcan be obtained by SPR assay. In some embodiments, the antibody binds specifically to PAD4 with a K of less than 5 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, from 0.01 nM to 5 nM, from 0.01 nM to 1 nM, from 0.05 nM to 1 nM, from 0.1 nM to 1 nM, from 0.1 nM to 0.5 nM, from 0.05 nM to 0.5 nM, from 0.05 nM to 0.1 nM, or from 0.5 nM to 1 nM in the presence of 1-2 mM calcium chloride (e.g., in the presence of 1 mM calcium chloride) in an SPR assay. D In some embodiments, the antibody binds specifically to PAD4 with a K of less than 5 nM, less than 1 nM, less than 0.5 nM, less than 0.1 nM, from 0.01 nM to 5 nM, from 0.01 nM to 1 nM, from 0.05 nM to 1 nM, from 0.1 nM to 1 nM, from 0.1 nM to 0.5 nM, from 0.05 nM to 0.5 nM, from 0.05 nM to 0.1 nM, or from 0.5 nM to 1 nM both in the presence of 1-2 mM calcium chloride (e.g., in the presence of 1 mM calcium chloride) and in the absence of calcium ions (absence of added calcium salt and presence of EDTA, e.g., 1 mM or 2 mM EDTA) by SPR. D In this way, in such cases, the antibody binds specifically to PAD4 both in the presence and absence of calcium ions such as calcium chloride. For example, see Tables 2 and 3 herein for examples.

[0291] In some embodiments, the anti-PAD4 antibody does not bind to human protein arginine deiminase 2 (PAD2). Thus, in a binding assay such as SPR, the binding to PAD2 is too weak to be detected. In some examples, the antibody specifically binds to human PAD4, but not to mouse PAD4 (i.e., the binding to mouse PAD4 is not detected in an SPR or similar assay), as described immediately above. In some examples, the antibody specifically binds to human PAD4, but not to cynomolgus PAD4 (i.e., the binding to cynomolgus PAD4 is not detected in an SPR assay), as described immediately above, while in other examples, the antibody specifically binds to both human and cynomolgus PAD4 in an SPR assay. In some examples, the antibody specifically binds to both human and cynomolgus PAD4 by SPR, but not to mouse PAD4. In other examples, the antibody specifically binds to human PAD4 as determined by SPR, but does not bind to cynomolgus or mouse PAD4.

[0292] In some embodiments, the antibody has an ECM score of less than 50, less than 30, less than 10, less than 5, 1-30, 1-20, 1-10, 1-5, 1-3, 1, 2, 3, 4, or 5 in an ECM assay (see, e.g., Table 5). As described in Example 5 below, the ECM score indicates the degree to which the antibody non-specifically binds to the extracellular matrix (ECM). The assay can be performed using a pre-coated ECM plate, such as a commercially available 96-well plate. The ECM score can be determined by incubating the antibody with the ECM-coated plate for 1 hour, then incubating with an HRP-conjugated detection antibody, reacting with a TMB substrate, and then dividing the absorbance value measured at 450 nm by the value of a control well without the added antibody (see Example 5 below).

[0293] In some embodiments, the antibodies herein have a retention time of 9 - 11 minutes at a flow rate of 1.0 ml / min and a column temperature of 25°C using a linear gradient of mobile phase A (0.1 M sodium phosphate pH 7.0, 2 M sodium sulfate) and mobile phase B (0.1 M sodium phosphate solution pH 7.0) over 20 minutes in hydrophobic interaction chromatography (HIC) on a TSKgel butyl-NPR column (4.6 mm × 3.5 cm, 2.5 μm particle size, Tosoh P / N 14947) in an Agilent 1260 Infinity II HPLC system (see Example 6 below). In some embodiments, the antibody has a melting temperature (Tm) between 60 - 70°C at both pH 6.0 and pH 8.3 as measured by intrinsic fluorescence. In some embodiments, the antibody has an aggregation temperature (Tagg) between 64 - 75°C at pH 6.0 and an aggregation temperature between 60 - 70°C at pH 8.3 as measured by static light scattering (see Example 7).

[0294] In some embodiments, the antibody has relatively low immunogenicity as measured using the Epivax® immunogenicity assay. A commercially available in silico immunogenic risk assessment algorithm [Epivax®] ranked peptide MHC class II binding for eight human HLA DRB1 allele supertypes to include >90% of the variation present in the human population (De Groot and Martin, Clin Immunol, 2009, 131(2): p. 189-201.) (see Example 15). For example, the in silico immunogenicity scores of the drugs Campath® (alemtuzumab), Rituxan® (rituximab), Zenapax® (daclizumab), Humicade® (CDP-571), Mylotarg® (gemtuzumab), and Avastin® (bevacizumab) range from 0% to 45% as shown in Table 10 below, and Campath® (alemtuzumab), Rituxan® (rituximab), Zenapax® (daclizumab), and Humicade® (CDP-571) are each in the range of 45% to 7%, respectively. In contrast, in some embodiments, the antibodies herein have immunogenicity scores in the range of 0 to 5%, such as 0 to 2%, or 0 to 1% in the same assay. Certain antibodies herein have an immunogenicity score of 0% (see Table 10). Thus, in certain instances, the antibody is less immunogenic than one, two, or all three of Campath® (alemtuzumab), Rituxan® (rituximab), and Zenapax® (daclizumab) as measured by an in silico immunogenicity assay.

[0295] In some embodiments, the antibody inhibits the enzymatic activity of PAD4 in vitro. For example, in some examples, the antibody inhibits the conversion of arginine to citrulline by PAD4 in the peptide substrate TSTGGRQGSHH (SEQ ID NO: 216) in vitro with an IC50 of 10 - 200 nM, 50 - 200 nM, 10 - 100 nM, 20 - 100 nM, or 50 - 100 nM (see Example 4, Table 4). For example, in some examples, the antibody inhibits the conversion of arginine to citrulline by PAD4 in the peptide substrate (SHQESTRGKSKGKAAAAA; SEQ ID NO: 232) in vitro. For example, in some examples, the antibody inhibits dose-dependently at an IC50 of 0.1 - 10 nM, for example 0.2 - 5 nM, at an appropriately 1 - 8 μg / mL PAD4 concentration (see, for example, Example 16 below).

[0296] 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 subject (e.g., a subject having a disease, e.g., a disease disclosed herein, e.g., an inflammatory disease or an autoimmune disease, e.g., a subject having rheumatoid arthritis). The endogenous anti-PAD4 antibodies can, for example, include or be 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 an endogenous antibody (activating antibody) that binds to and activates human PAD4. In some embodiments, the activating antibody is an antibody from a patient having rheumatoid arthritis. In some embodiments, the activating antibody is an antibody that is cross-reactive with respect to human PAD3 and human PAD4.

[0297] In some examples, the antibody reduces the amount of extracellular citrullinated histone H3 in LPS-stimulated human blood monocytes compared to an isotype control antibody. In some examples, the antibody reduces the secretion of GM-CSF in LPS-stimulated human blood monocytes compared to an isotype control antibody. In some examples, the antibody reduces the gene expression of GM-CSF in LPS-stimulated human blood monocytes compared to an isotype control antibody. In some examples, the antibody has two of these three properties. In some examples, 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 internalizing by LPS-stimulated human blood monocytes. For example, the antibody may be labeled with a dye that detects internalization by monocytes. In some examples, the antibody can be internalized by LPS-stimulated CD14+ human monocytes (see Example 18). Such results indicate that the anti-PAD4 antibodies herein can act intracellularly in monocytes and, in some embodiments, can block the function of PAD4 both extracellularly and intracellularly.

[0298] In some embodiments, the antibody inhibits PAD4 function in the inflamed lung. For example, this can be demonstrated by a decrease in citrullination of histone H3 or ITIH4 in bronchoalveolar lavage fluid (BALF) collected from the lung, as described in Examples 20 or 25 herein. For example, BALF can be collected from mice with acute or chronic lung inflammation and assayed for the amount of citrullinated H3 or ITIH4 protein in the presence of the antibody and in the presence of wild-type controls. In some embodiments, the mice are human PAD4 knock-in mice. In some examples, in human PAD4 knock-in mice, in the presence of the antibody, the amount of citrullinated H3 or ITIH4 is measured with respect to untreated (vehicle) mice, mice administered IC, and mice administered anti-PAD4 antibody, for example, by the following formula: % inhibition = [(IC - vehicle) - (PAD4 mAb - vehicle) / (IC - vehicle)] × 100. When calculated, the amount of citrullinated H3 is reduced by at least 15%, at least 30%, at least 40%, at least 50%, or at least 60% compared to an isotype control (IC) antibody, and / or the amount of citrullinated ITIH4 is reduced by at least 50%, at least 70%, or at least 80%.

[0299] In some embodiments, the antibody inhibits PAD4 function in joints with inflammation, as described, for example, in Examples 21, 22, and / or 26. This can be demonstrated by a decrease in citrullination of ITIH4 and / or PRG4 in joint tissue in a mouse model of acute or chronic joint inflammation induced, for example, by intra-articular injection of LPS. In some examples, the mouse can be a human PAD4 knock-in mouse. In some examples, the antibody reduces the amount of extracellular citrullinated PRG4 and / or citrullinated ITIH4 in the mouse patella in a mouse model of LPS-induced acute joint injury compared to an isotype control antibody. In some examples, the EC50 for the reduction of citrullinated ITIH4 and / or citrullinated PRG4 is 2 nM or less, 1 nM or less, 0.1 nM or less, 0.05 - 2 nM, or 0.1 - 1 nM.

[0300] In some embodiments, the antibodies herein can reduce pristane-induced extracellular trap-forming neutrophils (NETosis) and / or extracellular trap-forming monocytes (METosis) (see Example 23). METosis is the process by which extracellular traps consisting of cellular DNA and cellular proteins dotted with histones are released from monocytes or macrophages. This net-like material formed by either METosis or NETosis (neutrophil origin) is important in defense against microorganisms but is also a major promoter of autoimmune conditions and sterile inflammation. For example, pristane may be intraperitoneally injected into mice, such as human PAD4 knock-in mice, for example, after treatment with the antibodies herein or isotype control antibodies. In some examples, in such a pristane-induced mouse model, the antibodies herein can reduce citrullination of H3 in neutrophils, monocytes, M1 macrophages, and / or M2 macrophages in ascites as compared to isotype control antibodies. In further examples, the antibodies can also reduce the amount of soluble markers of neutrophils and monocytes / macrophages in mouse ascites, such as elastase, MPO, MIP-2 alpha, GRO alpha / KC, MCP1, MIP 1 beta, IL6, and MIP3 alpha (see Example 23).

[0301] In some embodiments, in a murine collagen-induced arthritis model, the antibodies herein may inhibit PAD4-dependent responses (see Example 24). For example, in some embodiments, the antibodies herein significantly reduce the arthritis clinical score of mice in an arthritis model as compared to an isotype control according to the following scale: (1) normal; (2) mild, with distinct erythema and swelling of the ankle or wrist, or distinct erythema and swelling limited to individual fingers, regardless of the number of affected fingers; (3) moderate erythema and swelling of the ankle or wrist; (4) severe erythema and swelling of the entire foot including the toes; (5) extremities with maximal inflammation involving multiple joints. For example, in some instances, the antibodies herein also reduce NETosis and METosis, as evaluated 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 citrulline in the H3 protein in neutrophils, monocytes, and / or macrophages (see Example 24).

[0302] In some embodiments, the antibody inhibits the citrullination of one or more of proteoglycan 4 (PRG4), fibrinogen A (FGA), inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), alpha-1-microglobulin / bikunin precursor (AMBP), and gelsolin (GSN) in serum both in the presence and absence of a disease-related anti-PAD4 antibody, as described, for example, in Example 27. In some...

Claims

1. An isolated antibody that specifically binds to the protein arginine deiminase 4 (PAD4), (a) A heavy chain variable region (VH) comprising a heavy chain complementarity determination region 1 (HCDR1) containing the amino acid sequence of SEQ ID NO: 62 and a heavy chain variable region (VL) comprising a light chain complementarity determination region 1 (LCDR1) containing the amino acid sequence of SEQ ID NO: 7; (b) A heavy chain variable region (VH) comprising a heavy chain complementarity determination region 1 (HCDR1) containing the amino acid sequence of SEQ ID NO: 62, an HCDR2 containing the amino acid sequence of SEQ ID NO: 5, and an HCDR3 containing the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determination region 1 (LCDR1) containing the amino acid sequence of SEQ ID NO: 7, an LCDR2 containing the amino acid sequence of SEQ ID NO: 8, and an LCDR3 containing the amino acid sequence of SEQ ID NO: 9; (c) 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; (d) A heavy chain variable region (VH) comprising a heavy chain complementarity determination region 1 (HCDR1) containing the amino acid sequence of SEQ ID NO: 62 and an HCDR3 containing the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determination region 1 (LCDR1) containing 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; (e) A heavy chain variable region (VH) comprising a heavy chain complementarity determination region 1 (HCDR1) containing the amino acid sequence of SEQ ID NO: 62, an HCDR2 containing the amino acid sequence of SEQ ID NO: 5, and an HCDR3 containing the amino acid sequence of SEQ ID NO: 6; and a light chain variable region (VL) comprising a light chain complementarity determination region 1 (LCDR1) containing the amino acid sequence of SEQ ID NO: 7, an LCDR2 containing the amino acid sequence of SEQ ID NO: 8, and an LCDR3 containing 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; or (f) 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. Isolated antibodies.

2. The isolated antibody according to claim 1, 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.

3. The isolated antibody according to claim 1, wherein VH contains glycine at position 94 of Kabat (corresponding to position 98 of SEQ ID NO: 68).

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

70.

5. An isolated antibody according to any one of claims 1 to 4, comprising SEQ ID NO: 217, or comprising SEQ ID NO: 217 and SEQ ID NO: 218, which binds to an epitope on PAD4.

6. An isolated antibody according to any one of claims 1 to 4, wherein the antibody is an IgA, IgG, or IgM antibody.

7. The isolated antibody according to claim 6, wherein the antibody is IgG and comprises a wild-type, human IgG1, IgG2, or IgG4 heavy chain constant region.

8. (a) comprising a human IgG1 or IgG2 heavy chain constant region comprising an amino acid sequence which is at least 90%, at least 95%, at least 97%, or at least 99% identical to any one of the amino acid sequences of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, and 192, and / or A light chain constant region containing 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, or (b) 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 any one of the amino acid sequences of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, and 192, and / or The light chain constant region includes 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. An isolated antibody according to any one of claims 1 to 4.

9. A human IgG1 or IgG2 heavy chain constant region containing one of the amino acid sequences of SEQ ID NOs: 174, 176, 178, 180, 182, 184, 186, 188, 190, and 192, and / or Including the light chain constant region containing the amino acid sequence of SEQ ID NO: 194, An isolated antibody according to any one of claims 1 to 4.

10. An isolated antibody according to any one of claims 1 to 4, which is a full-length antibody or an IgG antibody lacking C-terminal lysine in the heavy chain constant region.

11. Antibody fragments, e.g., Fv, single-stranded Fv (scFv), Fab, Fab', or (Fab') ２ The isolated antibody according to any one of claims 1 to 4.

12. An isolated antibody that specifically binds to the protein arginine deiminase 4 (PAD4), comprising a heavy chain (HC) containing 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) containing the amino acid sequence of SEQ ID NO: 70 followed by the amino acid sequence of SEQ ID NO:

194.

13. An isolated antibody that specifically binds to the protein arginine deiminase 4 (PAD4), comprising a heavy chain (HC) containing 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) containing the amino acid sequence of SEQ ID NO: 70 followed by the amino acid sequence of SEQ ID NO:

194.

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

200.

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

200.

16. An isolated antibody according to any one of claims 1, 4, or 12-15, which is a bispecific or polyspecific antibody, or is conjugated covalently or noncovalently to at least one other molecule, wherein the at least one other molecule optionally comprises a detection label and / or a drug.

17. An isolated antibody according to any one of claims 1, 4, or 12-15, which does not bind to human protein arginine deiminase 2 (PAD2).

18. The following features: (a) When measured using ELISA at an IC50 of 0.1–10 nM, for example 0.2–5 nM, it inhibits in vitro PAD4 citrullination of arginine in a peptide substrate (SHQESTRGKSKGKAAAAAAA; SEQ ID NO: 232) treated with recombinant human PAD4 at a concentration of 1 μg / mL–8 μg / mL; (b) Inhibits PAD4 deiminolysis of Nα-benzoyl-L-arginine ethyl hydrochloride (BAEE); (c) K less than 5 nM Ｄ It specifically binds to PAD4; (d) (i) at 37°C in the presence of 1 mM calcium chloride, and (ii) at 37°C with added Ca ２＋ In the absence of and in the presence of 2 mM EDTA, the K2 is 0.05 nM to 0.5 nM as determined by surface plasmon resonance (SPR). Ｄ It specifically binds to PAD4; (e) Having 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; (f) Lower immunogenicity than one, two, or all three of Campath® (aremtuzumab), Rituxan® (rituximab), and Zenapax® (daclizumab) as measured using the Epivax antibody immunogenicity scale in an in silico immunogenicity assay; (g) Compared to an isotype control antibody, it reduces one or more of the extracellular citrullinated histone H3 levels, GM-CSF secretion, and GM-CSF gene expression in LPS-stimulated human blood monocytes; (h) It is possible to move it into the body by LPS-stimulated human blood monocytes; (i) Inhibit PAD4 function in inflamed joints; (j) Reduce the amount of one or both of extracellular citrullinated H3 and citrullinated ITIH4 in BALF in a human PAD4 knock-in mouse LPS-induced acute pneumonia model compared to an isotype control antibody; and / or (k) Reduces the amount of one or both of extracellular citrullinated PRG4 and citrullinated ITIH4 in mouse patella in an LPS-induced acute joint injury model using human PAD4 knock-in mice, compared to isotype control antibodies. An isolated antibody according to any one of claims 1, 4, or 12-15, comprising one or more of the above.

19. A pharmaceutical composition comprising an isolated antibody and a pharmaceutically acceptable carrier according to any one of claims 1 to 18.

20. An isolated nucleic acid or a set of two or more nucleic acids encoding an isolated antibody according to any one of claims 1 to 18.

21. An isolated vector comprising one or more nucleic acids encoding the heavy chain and light chain of an isolated antibody according to any one of claims 1 to 18.

22. An isolated host cell comprising an isolated nucleic acid or a set of two or more nucleic acids as described in claim 20, or a vector comprising the isolated nucleic acid or a set of two or more nucleic acids.

23. A method for producing an isolated antibody that specifically binds to PAD4, comprising culturing the isolated host cells described in claim 22 under conditions suitable for the expression of the antibody, and optionally further comprising recovering the antibody from the host cells.

24. An isolated antibody produced by the method of claim 23.

25. An isolated antibody according to any one of claims 1 to 18, for use in the treatment of autoimmune disorders.

26. The isolated antibody for use according to claim 25, wherein the autoimmune disorder is rheumatoid arthritis (RA), 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).

27. ​​The isolated antibody for use according to claim 25, wherein the autoimmune disorder is rheumatoid arthritis (RA).

28. An isolated antibody for use according to claim 25, wherein at least one further therapeutic agent is administered, the at least one further therapeutic agent being one or more of methotrexate, adalimumab, etanercept, infliximab, hydroxychloroquine, sulfasalazine, leflunomide, abatacept, anakinra, certolizumab, golimumab, rituximab, sarilumab, tocilizumab, baricitinib, tofacitinib, and upadacitinib.

29. The pharmaceutical composition according to claim 19 for use in the treatment of autoimmune disorders.

30. The pharmaceutical composition for use according to claim 29, wherein the autoimmune disorder is rheumatoid arthritis (RA), 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).

31. The pharmaceutical composition for use according to claim 29, wherein the autoimmune disorder is rheumatoid arthritis (RA).

32. An isolated antibody according to any one of claims 1 to 18, for use in the treatment of inflammatory bowel disease (IBD).

33. An isolated antibody for use according to claim 32, wherein the IBD is ulcerative colitis or Crohn's disease.

34. An isolated antibody according to any one of claims 1 to 18, for use in the treatment of subjects at risk of developing rheumatoid arthritis (RA).

35. The above subject is subject to the following conditions: a. At least one first-degree relative (e.g., parent or sibling) who has RA; b. Presence of anti-citrullinated protein antibodies (ACPA) in serum; c. Presence of rheumatoid factor (RF) in the serum; d. Joint pain in at least one joint; e. The presence of inflammation in at least one joint as observed by ultrasound or magnetic resonance imaging (MRI); and f. Unclassifiable arthritis An isolated antibody for use according to claim 34, having one or more of the following:

36. The isolated antibody for use according to claim 34, wherein the subject is in remission.

37. The isolated antibody for use according to claim 34, wherein the treatment is for delaying the onset of RA.

38. An isolated antibody according to any one of claims 1 to 18, for use in inhibiting NETosis or METosis in a target.

39. An isolated antibody according to any one of claims 1 to 18, for use in inhibiting citrullination in a target.

40. An isolated antibody according to any one of claims 1 to 18, for use in therapeutic purposes.

41. An in vitro method for inhibiting NETosis or METosis in a biological sample, comprising administering an effective amount of an isolated antibody according to any one of claims 1 to 18 to the biological sample.

42. An in vitro method for inhibiting citrullination in a biological sample, comprising administering an effective amount of an isolated antibody according to any one of claims 1 to 18 to the biological sample.

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