CNX antigen-binding molecules

Abstract

Antigen binding molecules capable of binding to calnexin (CNX), comprising (i) a CNX-binding moiety and (ii) a linker-payload moiety are disclosed herein. Also disclosed are compositions comprising such antigen binding molecules. Uses and methods involving antigen binding molecules capable of binding to calnexin (CNX) are also disclosed.

Description

[0001] CNX Antigen-Binding Molecules

[0002] This application claims priority from US 63 / 614,286 filed 22 December 2023, the contents and elements of which are incorporated by reference for all purposes.

[0003] Technical Field

[0004] The present disclosure relates to the fields of molecular biology, more specifically antibody technology and antibody-drug conjugates (ADCs). The present disclosure also relates to methods of medical treatment and prophylaxis.

[0005] Background

[0006] CNX is an endoplasmic reticulum (ER)-resident lectin chaperone protein, which binds to N-glycoproteins bearing monoglucosylated glycans, and recruits various other chaperones that mediate protein disulfide formation, proline isomerisation, and protein folding.

[0007] Recent studies have implicated CNX and CNX-containing complexes (e.g. CNX:ERp57) in the pathology of diseases / conditions including cancers, particularly through their ECM degrading activity (see Ros et al. Nat. Cell Biol. (2020) 22(11 ):1371 -1381 ). CNX:ERp57 complexes have also been shown to translocate to the surface of cancer cells, where they reduce disulfide bridges in the extracellular matrix (Ros et al., Nat. Cell Biol. 22, 1371-1381, 2020).

[0008] Ros et al. Nat. Cell Biol. (2020) 22(11 ): 1371 -1381 discloses anti-CNX antibodies ab10286 and ab22595. Abcam’s ab10286 and ab22595 are each rabbit polyclonal antibody preparations to human CNX. There remains a need to develop CNX-targeted therapeutics suitable for use in methods of medical treatment and prophylaxis.

[0009] Antibody-drug conjugates (ADCs) typically comprise a monoclonal antibody linked to a biologically active payload (e.g. a cytotoxic molecule) via a chemical linker. ADCs combine both the advantages of highly specific targeting ability and highly potent killing effect to achieve accurate and efficient elimination of target cells (e.g. cancer or disease-associated cells).

[0010] Summary

[0011] In a first aspect, the present disclosure provides an antigen-binding molecule that binds to CNX, comprising (i) a CNX-binding moiety and (ii) a linker-payload moiety.

[0012] In some embodiments, the CNX-binding moiety comprises:

[0013] (a)

[0014] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0015] HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and

[0016] (II) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; or

[0017] (b)

[0018] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0019] HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and

[0020] (II) a light chain variable (VL) region incorporating the following CDRs:

[0021] LC-CDR1 having the amino acid sequence of SEQ ID NO:179 LC-CDR2 having the amino acid sequence of SEQ ID NQ:180 LC-CDR3 having the amino acid sequence of SEQ ID NO:173; or

[0022] (c)

[0023] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0024] HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and

[0025] (II) a light chain variable (VL) region incorporating the following CDRs:

[0026] LC-CDR1 having the amino acid sequence of SEQ ID NQ:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or

[0027] (d)

[0028] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0029] HC-CDR1 having the amino acid sequence of SEQ ID NO:18 HC-CDR2 having the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NQ:20; and

[0030] (II) a light chain variable (VL) region incorporating the following CDRs:

[0031] LC-CDR1 having the amino acid sequence of SEQ ID NO:25 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:27; or

[0032] (e)

[0033] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0034] HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:49; and

[0035] (II) a light chain variable (VL) region incorporating the following CDRs:

[0036] LC-CDR1 having the amino acid sequence of SEQ ID NO:53 LC-CDR2 having the amino acid sequence of SEQ ID NO:54 LC-CDR3 having the amino acid sequence of SEQ ID NO:55; or

[0037] (f)

[0038] (I) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:62 HC-CDR3 having the amino acid sequence of SEQ ID NO:63; and

[0039] (II) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:68 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:69; or

[0040] (g)

[0041] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0042] HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:62 HC-CDR3 having the amino acid sequence of SEQ ID NO:63; and

[0043] (II) a light chain variable (VL) region incorporating the following CDRs:

[0044] LC-CDR1 having the amino acid sequence of SEQ ID NO:73 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:74; or

[0045] (h)

[0046] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0047] HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:62 HC-CDR3 having the amino acid sequence of SEQ ID NO:63; and

[0048] (II) a light chain variable (VL) region incorporating the following CDRs:

[0049] LC-CDR1 having the amino acid sequence of SEQ ID NO:78 LC-CDR2 having the amino acid sequence of SEQ ID NO:79 LC-CDR3 having the amino acid sequence of SEQ ID NQ:80; or

[0050] (i)

[0051] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0052] HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:83; and

[0053] (II) a light chain variable (VL) region incorporating the following CDRs:

[0054] LC-CDR1 having the amino acid sequence of SEQ ID NO:73 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:74; or

[0055] (j)

[0056] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0057] HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:86; and

[0058] (II) a light chain variable (VL) region incorporating the following CDRs:

[0059] LC-CDR1 having the amino acid sequence of SEQ ID NO:89 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NQ:90; or (k)

[0060] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0061] HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:95 HC-CDR3 having the amino acid sequence of SEQ ID NO:96; and

[0062] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0063] LC-CDR1 having the amino acid sequence of SEQ ID NQ:101 LC-CDR2 having the amino acid sequence of SEQ ID NQ:102 LC-CDR3 having the amino acid sequence of SEQ ID NQ:103; or

[0064] (l)

[0065] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0066] HC-CDR1 having the amino acid sequence of SEQ ID NQ:108 HC-CDR2 having the amino acid sequence of SEQ ID NQ:109 HC-CDR3 having the amino acid sequence of SEQ ID NQ:110; and

[0067] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0068] LC-CDR1 having the amino acid sequence of SEQ ID NO:115 LC-CDR2 having the amino acid sequence of SEQ ID NO:116 LC-CDR3 having the amino acid sequence of SEQ ID NO:117; or

[0069] (m)

[0070] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0071] HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:122; and

[0072] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0073] LC-CDR1 having the amino acid sequence of SEQ ID NO:125 LC-CDR2 having the amino acid sequence of SEQ ID NO:126 LC-CDR3 having the amino acid sequence of SEQ ID NO:127; or

[0074] (n)

[0075] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0076] HC-CDR1 having the amino acid sequence of SEQ ID NO:132 HC-CDR2 having the amino acid sequence of SEQ ID NO:133 HC-CDR3 having the amino acid sequence of SEQ ID NO:134; and

[0077] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0078] LC-CDR1 having the amino acid sequence of SEQ ID NO:139 LC-CDR2 having the amino acid sequence of SEQ ID NQ:140 LC-CDR3 having the amino acid sequence of SEQ ID NQ:80; or

[0079] (o)

[0080] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0081] HC-CDR1 having the amino acid sequence of SEQ ID NO:146 HC-CDR2 having the amino acid sequence of SEQ ID NO:147 HC-CDR3 having the amino acid sequence of SEQ ID NO:148; and

[0082] (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:153; or (P)

[0083] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0084] HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:156; and

[0085] (II) a light chain variable (VL) region incorporating the following CDRs:

[0086] LC-CDR1 having the amino acid sequence of SEQ ID NO:158 LC-CDR2 having the amino acid sequence of SEQ ID NO:159 LC-CDR3 having the amino acid sequence of SEQ ID NQ:160; or

[0087] (q)

[0088] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0089] HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and

[0090] (II) a light chain variable (VL) region incorporating the following CDRs:

[0091] LC-CDR1 having the amino acid sequence of SEQ ID NO:171 LC-CDR2 having the amino acid sequence of SEQ ID NO:172 LC-CDR3 having the amino acid sequence of SEQ ID NO:173; or

[0092] (r)

[0093] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0094] HC-CDR1 having the amino acid sequence of SEQ ID NO:185 HC-CDR2 having the amino acid sequence of SEQ ID NO:186 HC-CDR3 having the amino acid sequence of SEQ ID NO:187; and

[0095] (II) a light chain variable (VL) region incorporating the following CDRs:

[0096] LC-CDR1 having the amino acid sequence of SEQ ID NO:73 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:194; or

[0097] (s)

[0098] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0099] HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:199 HC-CDR3 having the amino acid sequence of SEQ ID NQ:200; and

[0100] (II) a light chain variable (VL) region incorporating the following CDRs:

[0101] LC-CDR1 having the amino acid sequence of SEQ ID NQ:205 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NQ:206; or

[0102] (t)

[0103] (I) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:211

[0104] HC-CDR3 having the amino acid sequence of SEQ ID NO:212; and

[0105] (II) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:216 LC-CDR2 having the amino acid sequence of SEQ ID NO:172 LC-CDR3 having the amino acid sequence of SEQ ID NO:217; or

[0106] (u)

[0107] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0108] HC-CDR1 having the amino acid sequence of SEQ ID NO:222 HC-CDR2 having the amino acid sequence of SEQ ID NO:223 HC-CDR3 having the amino acid sequence of SEQ ID NO:224; and

[0109] (II) a light chain variable (VL) region incorporating the following CDRs:

[0110] LC-CDR1 having the amino acid sequence of SEQ ID NO:229 LC-CDR2 having the amino acid sequence of SEQ ID NO:172 LC-CDR3 having the amino acid sequence of SEQ ID NQ:230; or

[0111] (v)

[0112] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0113] HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:199 HC-CDR3 having the amino acid sequence of SEQ ID NQ:200; and

[0114] (II) a light chain variable (VL) region incorporating the following CDRs:

[0115] LC-CDR1 having the amino acid sequence of SEQ ID NO:235 LC-CDR2 having the amino acid sequence of SEQ ID NO:236 LC-CDR3 having the amino acid sequence of SEQ ID NO:237; or

[0116] (w)

[0117] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0118] HC-CDR1 having the amino acid sequence of SEQ ID NO:185 HC-CDR2 having the amino acid sequence of SEQ ID NO:243 HC-CDR3 having the amino acid sequence of SEQ ID NO:244; and

[0119] (II) a light chain variable (VL) region incorporating the following CDRs:

[0120] LC-CDR1 having the amino acid sequence of SEQ ID NO:248 LC-CDR2 having the amino acid sequence of SEQ ID NO:249 LC-CDR3 having the amino acid sequence of SEQ ID NQ:250.

[0121] In some embodiments, the CNX-binding moiety comprises: a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32, 165, 1 , 17, 47, 60, 82, 85, 94, 107, 121 , 131 , 154, 155, 184, 198, 210, 221 or 242; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:40, 178, 9, 24, 52, 67, 72, 77, 88, 100, 114, 124, 138, 152, 157, 170, 191, 204, 215, 228, 234 or 247. In some embodiments, the CNX-binding moiety comprises:

[0122] (i) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:40; or

[0123] (ii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:165; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:178; or

[0124] (iii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:1; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:9; or

[0125] (iv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:17; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:24; or

[0126] (v) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:47; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:52; or

[0127] (vi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:60; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:67; or

[0128] (vii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:60; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:72; or

[0129] (viii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:60; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:77; or (ix) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:82; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:72; or

[0130] (x) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:85; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:88; or

[0131] (xi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:94; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:100; or

[0132] (xii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:107; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:114; or

[0133] (xiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:121 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:124; or

[0134] (xiv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:131 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:138; or

[0135] (xv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:145; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:152; or

[0136] (xvi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:155; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:157; or

[0137] (xvii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:165; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:170; or

[0138] (xviii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:184; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:191; or

[0139] (xix) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:198; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:204; or

[0140] (xx) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:210; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:215; or

[0141] (xxi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:221 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:228; or

[0142] (xxii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:198; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:234; or

[0143] (xxiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:242; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:247.

[0144] In some embodiments, the linker-payload moiety comprises or consists of a cytotoxic agent.

[0145] In some embodiments, the linker-payload moiety comprises or consists of: a microtubule-targeting agent, a DNA-targeting agent, an RNA-targeting agent, an immune system-activating agent, an apoptosispromoting agent, a metabolism-inhibiting agent and a proteasome inhibiting agent.

[0146] In some embodiments, the linker-payload moiety comprises or consists of a microtubule-targeting agent.

[0147] In some embodiments, the linker-payload moiety comprises or consists of an auristatin. In some embodiments, the linker-payload moiety comprises or consists of monomethyl auristatin E (MMAE) or auristatin F.

[0148] In some embodiments, the linker-payload moiety comprises or consists of a DNA-targeting agent. In some embodiments, the linker-payload moiety comprises or consists of a topoisomerase inhibitor, optionally wherein the topoisomerase inhibitor is a TOP1 inhibitor. In some embodiments, the linkerpayload moiety comprises or consists of camptothecin or a derivative thereof. In some embodiments, the linker-payload moiety comprises or consists of deruxtecan (Dxd).

[0149] In some embodiments, the linker-payload moiety comprises cathepsin-cleavable linker. In some embodiments, the linker-payload moiety comprises a valine-citruline linker or a glycine-glycine- phenylalanine-glycine linker.

[0150] In some embodiments, the antigen-binding molecule is selected from one of: (a) wherein ABM is an antigen-binding moiety which comprises:

[0151] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0152] HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and

[0153] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0154] LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; wherein ABM is an antigen-binding moiety which comprises:

[0155] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0156] HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and

[0157] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0158] LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; or

[0159] (c) wherein SNAP is a SNAP protein, and wherein ABM is an antigen-binding moiety which comprises:

[0160] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0161] HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and

[0162] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0163] LC-CDR1 having the amino acid sequence of SEQ ID NO:179

[0164] LC-CDR2 having the amino acid sequence of SEQ ID NQ:180

[0165] LC-CDR3 having the amino acid sequence of SEQ ID NO:173.

[0166] In some embodiments, the drug to antibody ratio (DAR) is between 1 and 8.

[0167] The present disclosure also provides a composition comprising an antigen-binding molecule according to the present disclosure, and a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0168] The present disclosure also provides an antigen-binding molecule according to the present disclosure, or a composition according to the present disclosure, for use as a medicament.

[0169] The present disclosure also provides an antigen-binding molecule according to the present disclosure, or a composition according to the present disclosure, for use in a method of medical treatment or prophylaxis, wherein the disease is characterised by cells which exhibit increased levels of GALA activation and / or increased cell surface expression of CNX. The present disclosure also provides an antigen-binding molecule according to the present disclosure, or a composition according to the present disclosure, for use in treating or preventing a cancer.

[0170] The present disclosure also provides the use of an antigen-binding molecule according to the present disclosure, or a composition according to the present disclosure, in the manufacture of a medicament for treating or preventing a cancer.

[0171] The present disclosure also provides a method of treating or preventing a cancer, comprising administering to a subject a therapeutically- or prophylactically-effective amount of an antigen-binding molecule according to the present disclosure, or a composition according to the present disclosure.

[0172] In some embodiments, the cancer is selected from: liver cancer, breast cancer, oral cancer, oral squamous cell carcinoma, sarcoma, lung cancer, prostate cancer, bladder cancer, renal cancer, melanoma, pancreatic cancer, endometrial cancer, colorectal cancer, ovarian cancer, cervical cancer, brain cancer, bile duct cancer, testicular cancer, and thyroid cancer.

[0173] The present disclosure also provides the use of antigen-binding molecule according to the present disclosure to deplete cells expressing CNX or increase killing of cells expressing CNX.

[0174] The present disclosure also provides an in vitro complex, optionally isolated, comprising an antigenbinding molecule according to the present disclosure bound to CNX.

[0175] The present disclosure also provides a method comprising contacting a cell expressing or suspected of expressing CNX with an antigen-binding molecule that binds to CNX, wherein the antigen binding molecule comprises (i) a CNX-binding moiety and (ii) a linker-payload moiety, wherein the antigen-binding molecule kills the cell.

[0176] The present disclosure also provides a method of killing a cell expressing or suspected of expressing CNX, said method comprising contacting the cell with an antigen-binding molecule that binds to CNX, wherein the antigen binding molecule comprises (i) a CNX-binding moiety and (ii) a linker-payload moiety.

[0177] In some embodiments, the cell is a cancer cell. In some embodiments, the cancer cell is a liver cancer cell, a pancreatic cancer cell or a breast cancer cell.

[0178] Description

[0179] The present disclosure relates to antigen-binding molecules comprising a CNX-binding moiety, optionally further comprising a linker-payload moiety.

[0180] CNX and CRT

[0181] The present disclosure relates to CNX-specific antigen-binding molecules. Human CNX (also known as CNX, CANX or I P90) is the protein identified by UniProt P27824. Alternative splicing of mRNA encoded by the human CANXgene yields three main CNX isoforms: isoform 1 (SEQ ID NO:333), isoform 2 (SEQ ID NO:334) and isoform 3 (SEQ ID NO:335). Isoform 2 differs from isoform 1 by insertion of a 35 amino acid sequence after position 1 of SEQ ID NO:333. Positions 1 to 108 of SEQ ID NO:333 are absent from isoform 3.

[0182] Human CNX isoform 1 comprises an N-terminal signal peptide (SEQ ID NO:336), followed by a calcium- binding lumenal domain (SEQ ID NO:337), a single-pass transmembrane domain (SEQ ID NO:338) and an acidic cytoplasmic domain (SEQ ID NO:339) at the C-terminus. The lumenal domain comprises a globular lectin domain (SEQ ID NQ:340), followed by an arm-like, proline-rich P-domain (SEQ ID NO:341) and a second lectin domain (SEQ ID NO:342). The mature form of human CNX isoform 1 is shown in SEQ ID NO:343.

[0183] In this specification ‘CNX’ refers to CNX from any species, and includes isoforms, fragments, variants or homologues from any species. In some embodiments CNX is CNX from a mammal (e.g. a therian, placental, epitherian, preptotheria, archontan, primate (rhesus, cynomolgous, non-human primate or human)). In some embodiments, the CNX is human CNX or mouse CNX.

[0184] As used herein, a ‘fragment’, ‘variant’, ‘isoform’ or ‘homologue’ of a given protein may optionally be characterised as having at least 60%, preferably one of 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater amino acid sequence identity to the amino acid sequence of the reference protein (e.g. a reference isoform).

[0185] A ‘fragment’ generally refers to a fraction of the reference protein. A ‘variant’ generally refers to a protein having an amino acid sequence comprising one or more amino acid substitutions, insertions, deletions or other modifications relative to the amino acid sequence of the reference protein but retaining a considerable degree of sequence identity (e.g. at least 60%) to the amino acid sequence of the reference protein. An ‘isoform’ generally refers to a variant of the reference protein expressed by the same species as the species of the reference protein (e.g. human CNX isoform 1 , isoform 2 and isoform 3 are all isoforms of one another). A ‘homologue’ generally refers to a variant of the reference protein produced by a different species as compared to the species of the reference protein. For example, human CNX isoform 1 (UniProt: P27824-1, v2; SEQ ID NO:333) and mouse CNX (UniProt: P35564-1 , v1 ; SEQ ID NO:344) are homologues of one another. Homologues include orthologues.

[0186] Isoforms, fragments, variants or homologues of CNX according to the present disclosure may optionally be characterised as having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to the amino acid sequence of an immature or mature CNX isoform from a given species, e.g. human.

[0187] Isoforms, fragments, variants or homologues may optionally be functional isoforms, fragments, variants or homologues, e.g. having a functional property / activity of the reference CNX (e.g. human CNX isoform 1 ), as determined by analysis by a suitable assay for the functional property / activity. For example, an isoform, fragment, variant or homologue of CNX may display binding to a monoglucosylated glycan- bearing N-glycoprotein, and / or association with ERp57, cyclophilin B and / or ERp29.

[0188] In some embodiments, the CNX comprises, or consists of, an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:333, 334, 335 or 343.

[0189] In some embodiments, the CNX comprises, or consists of, an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:344 or 352.

[0190] A ‘fragment’ of a reference protein may be of any length (by number of amino acids), although may optionally be at least 25% of the length of the reference protein (that is, the protein from which the fragment is derived) and may have a maximum length of one of 50%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the length of the reference protein.

[0191] A fragment of CNX may have a minimum length of one of 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550 or 600 amino acids, and may have a maximum length of one of 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550 or 600 amino acids.

[0192] In some embodiments, a fragment of CNX comprises, or consists of, an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:343, 337, 338, 339, 340, 341 or 342.

[0193] In some embodiments, a fragment of CNX comprises, or consists of, an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:352, 346, 347, 348, 349, 350 or 351.

[0194] In some embodiments, the antigen-binding molecules of the present disclosure display binding to Calreticulin (CRT).

[0195] Inventors observed that CNX-depleted cells can compensate for CNX loss through the action of CRT. Therefore, the inventors identified antigen-binding molecules capable of binding both CNX and CRT.

[0196] In some embodiments, the antigen-binding molecule is cross-reactive for human CNX and CRT.

[0197] As used herein, a ‘cross-reactive’ antigen-binding molecule / moiety / domain / polypeptide binds to the target antigens for which the antigen-binding molecule / moiety / domain is cross-reactive. For example, an antigen-binding molecule / moiety / domain / polypeptide which is cross-reactive for CNX and CRT binds to CNX and is also capable of binding to CRT. Cross-reactive antigen-binding molecules / moieties / domains / polypeptides may display specific binding to each of the target antigens. Human CRT (also known as calreticulin, calregulin or ERp60) is the protein identified by UniProt P27797. Human CRT has the amino acid sequence shown in SEQ ID NO:353. Human CRT comprises an N- terminal signal peptide (SEQ ID NO:354), followed by a calcium-binding N-domain (SEQ ID NO:355), and an acidic C-domain (SEQ ID NO:356) at the C-terminus. The N-domain comprises a globular lectin domain (SEQ ID NO:357), followed by an arm-like, proline-rich P-domain (SEQ ID NO:359) and a second lectin domain (SEQ ID NO:358). The mature form of human CRT is shown in SEQ ID NQ:360.

[0198] In this specification ‘CRT’ refers to CRT from any species, and includes isoforms, fragments, variants or homologues from any species. In some embodiments CRT is CRT from a mammal (e.g. a therian, placental, epitherian, preptotheria, archontan, primate (rhesus, cynomolgous, non-human primate or human)). In some embodiments, the CRT is human CRT or mouse CRT.

[0199] Isoforms, fragments, variants or homologues of CRT according to the present disclosure may optionally be characterised as having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to the amino acid sequence of an immature or mature CRT isoform from a given species, e.g. human.

[0200] Isoforms, fragments, variants or homologues of CRT may optionally be functional isoforms, fragments, variants or homologues, e.g. having a functional property / activity of the reference CRT (e.g. human CRT), as determined by analysis by a suitable assay for the functional property / activity. For example, an isoform, fragment, variant or homologue of CRT may display binding to a monoglucosylated glycan- bearing N-glycoprotein, and / or association with ERp57, cyclophilin B and / or ERp29.

[0201] In some embodiments, the CRT comprises, or consists of, an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:353 or 360.

[0202] A fragment of CRT may have a minimum length of one of 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, or 400 amino acids, and may have a maximum length of one of 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, or 400 amino acids.

[0203] In some embodiments, a fragment of CRT comprises, or consists of, an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NQ:360, 355, 356, 357, 358 or 359.

[0204] The structure and function of CNX and CRT is reviewed e.g. in Kozlov and Gehring, FEBS J. (2020) 287(20):4322-4340, which is hereby incorporated by reference in its entirety.

[0205] CNX and CRT are endoplasmic reticulum (ER)-resident lectin chaperone proteins. CNX / CRT bind N-glycoproteins bearing monoglucosylated glycans and recruit various other chaperones which mediate protein disulfide formation, proline isomerisation, and protein folding. CNX / CRT are able to associate with the protein folding enzyme ERp57 to catalyse glycoprotein-specific disulfide bond formation. CNX:ERp57 complexes have also been shown to translocate to the surface of cancer cells, where they reduce disulfide bridges in the extracellular matrix (Ros et al., Nat. Cell Biol. 22, 1371-1381 , 2020). The reduction of disulfide bridges has been shown to be essential for the effective activity of matrix metalloproteinases (MMPs), and thus for the degradation of the extracellular matrix in cancer. CNX / CRT also associate with the peptidyl-prolyl cis-trans isomerase cyclophilin B (CypB), for the proline isomerisation of peptide bonds. CNX / CRT have also been reported to associate with ERp29 to form CNX / CRT:ERp29 complexes, which have a general chaperone function. CNX also functions as a chaperone for the folding of MHC class I a-chain in the membrane of the ER.

[0206] Processing by glucosidase II removes the glucose residue of the monoglucosylated N-glycan required for interaction of the glycoprotein with CNX / CRT, resulting in liberation of the mature, processed glycoprotein from CNX / CRT. For proteins that have not yet folded properly, UDP-glucose:glycoprotein glucosyltransferase (UGGT) acts as a checkpoint by re-adding a glucose residue back onto the N-glycan, reconstituting the interaction site for CNX / CRT. In this way, misfolded proteins re-associate with CNX / CRT for additional rounds of chaperone-mediated refolding, and their exit from the ER and progression to the Golgi is prevented. If multiple folding cycles are unsuccessful, terminally misfolded proteins are transported to the cytoplasm for degradation via the ER-associated protein degradation (ERAD) pathway.

[0207] Antigen-binding molecules and antigen-binding moieties

[0208] The present disclosure provides antigen-binding molecules capable of binding to CNX.

[0209] An ‘antigen-binding molecule’ refers to a molecule that binds to a given target antigen. Antigen-binding molecules comprise an antigen-binding moiety through which the antigen-binding molecule binds to its target antigen. The antigen-binding molecules of the present disclosure comprise an antigen-binding moiety that binds to CNX ( / .e. a CNX-binding moiety). In some embodiments, the antigen-binding molecules of the present disclosure comprise an antigen-binding moiety that binds to CNX and CRT.

[0210] Antigen-binding moieties may comprise, or may be derived from, antibodies ( / .e. immunoglobulins (Igs)) and antigen-binding fragments thereof. As used herein, ‘antibodies’ include monoclonal antibodies, polyclonal antibodies, monospecific and multispecific (e.g., bispecific, trispecific, etc.) antibodies, and antibody-derived antigen-binding molecules such as scFv, scFab, diabodies, triabodies, scFv-Fc, minibodies, single domain antibodies (e.g. VhH), etc.). Antigen-binding fragments of antibodies include e.g. Fv, Fab, F(ab’)2 and F(ab’) fragments. In some embodiments, an antigen-binding moiety may be an antibody or an antigen-binding fragment thereof (e.g. an scFv).

[0211] In some embodiments, the moiety capable of binding to a target antigen comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) of an antibody capable of specific binding to the target antigen.

[0212] The skilled person is readily able to produce antigen-binding moieties that bind to a given target antigen, in view of techniques that are well-known in the art. For example, Park and Smolen, Advances in Protein Chemistry (2001 ) 56: 369-421 describes approaches to the production of monoclonal antibodies suitable for use in humans, including raising xenogeneic antibodies and their subsequent humanisation, identification by human antibody gene-phage display, and production of antibodies in transgenic mice harbouring human antibody genes.

[0213] As used herein, a ‘peptide’ refers to a chain of two or more amino acid monomers linked by peptide bonds. A peptide typically has a length in the region of about 2 to 50 amino acids. A ‘polypeptide’ is a polymer chain of two or more peptides. Polypeptides typically have a length greater than about 50 amino acids.

[0214] The antigen-binding molecules of the present disclosure generally comprise an antigen-binding moiety comprising a VH and a VL of an antibody capable of specific binding to the target antigen (e.g. CNX). The antigen-binding domain formed by a VH and a VL may also be referred to herein as an Fv region.

[0215] An antigen-binding moiety may be, or may comprise, an antigen-binding polypeptide, or an antigenbinding polypeptide complex. An antigen-binding moiety may comprise more than one polypeptide which together form an antigen-binding moiety. The polypeptides may associate covalently or non-covalently. In some embodiments, the polypeptides form part of a larger polypeptide comprising the polypeptides (e.g. in the case of scFv comprising VH and VL, or in the case of scFab comprising VH-CH1 and VL-CL).

[0216] In some embodiments, an antigen-binding moiety according to the present disclosure comprises, or consists of, a polypeptide complex formed by proteimprotein interaction between constituent peptides / polypeptides of the antigen-binding moiety. An antigen-binding moiety may refer to a non- covalent or covalent complex of more than one polypeptide (e.g. 2, 3, 4, 6, or 8 polypeptides), e.g. an IgG-like antigen-binding moiety comprising two heavy chain polypeptides and two light chain polypeptides.

[0217] The antigen-binding moieties of the present disclosure may be designed and prepared using the sequences of monoclonal antibodies (mAbs) capable of binding to CNX. Antigen-binding regions of antibodies, such as single chain variable fragment (scFv), Fab and F(ab’)2 fragments may also be used / provided. An ‘antigen-binding region’ is any fragment of an antibody that binds to the target for which the given antibody is specific.

[0218] Antibodies generally comprise six complementarity-determining regions CDRs; three in the heavy chain variable (VH) region: HC-CDR1 , HC-CDR2 and HC-CDR3, and three in the light chain variable (VL) region: LC-CDR1 , LC-CDR2, and LC-CDR3. The six CDRs together define the paratope of the antibody, which is the part of the antibody that binds to the target antigen.

[0219] The VH region and VL region comprise framework regions (FRs) either side of each CDR, which provide a scaffold for the CDRs. From N-terminus to C-terminus, VH regions comprise the following structure: N term-[HC-FR1]-[HC-CDR1]-[HC-FR2]-[HC-CDR2]-[HC-FR3]-[HC-CDR3]-[HC-FR4]-C term; and VL regions comprise the following structure: N term-[LC-FR1]-[LC-CDR1]-[LC-FR2]-[LC-CDR2]-[LC-FR3]- [LC-CDR3]-[LC-FR4]-C term.

[0220] There are several different conventions for defining antibody CDRs and FRs, such as those described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991), Chothia et al., J. Mol. Biol. 196:901-917 (1987), and VBASE2, as described in Retter et al., NucL Acids Res. (2005) 33 (suppl 1 ): D671-D674. The CDRs and FRs of the VH regions and VL regions of the antibody clones described herein were defined according to the international IMGT (ImMunoGeneTics) information system (LeFranc et al., Nucleic Acids Res. (2015) 43 (Database issue):D413-22), which uses the IMGT V-DOMAIN numbering rules as described in Lefranc et al., Dev. Comp. Immunol. (2003) 27:55-77. In preferred embodiments, the CDRs and FRs of antigenbinding molecules referred to herein are defined according to the IMGT information system.

[0221] The antigen-binding molecules of the present disclosure comprise an antigen-binding moiety that binds to CNX.

[0222] In some embodiments, the antigen-binding moiety comprises the CDRs of an antigen-binding moiety that binds to CNX. In some embodiments, the antigen-binding moiety comprises the FRs of an antigenbinding moiety that binds to CNX. In some embodiments, the antigen-binding moiety comprises the CDRs and the FRs of an antigen-binding moiety that binds to CNX. That is, in some embodiments, the antigenbinding moiety comprises the VH region and the VL region of an antigen-binding moiety that binds to CNX.

[0223] In some embodiments, the antigen-binding moiety comprises or consists of monoclonal antibody clone AF18 (Invitrogen Cat. No. MA3-027), clone AF8 (Merck Cat. No. MABF2067), clone TO-5 (Merck Cat. No. C7617), clone 3H4A7 (Invitrogen Cat. No. MA5-15389), clone ARC0648 (Invitrogen Cat. No. MAS- 35588), clone GT1563 (GeneTex Cat. No. GTX629976), clone CANX / 1541 (GeneTex Cat. No. GTX34446), clone IE2.1C12 (Novus Biologicals Cat No. NBP2-36571), clone 1C2.2D11 (Novus Biologicals Cat No. NBP2-36570SS), clone 2A2C6 (Proteintech Cat. No. 66903-1-lg), clone C5C9 (Cell Signaling Technology, Inc Cat. No. 2679), clone E-10 (Santa Cruz Biotechnology Cat No. sc-46669), polyclonal antibodies ab10286 and ab22595 (Abeam), and anti-CNX antibodies disclosed in CN 101659702 A (e.g. the antibody produced by hybridoma CGMCC No. 3240).

[0224] In some embodiments, the antigen-binding moiety comprises the CDRs, FRs and / or the VH and / or VL regions of a CNX-binding antibody clone described herein, or CDRs, FRs and / or VH and / or VL regions which are derived from those of a CNX-binding antibody clone described herein. In some embodiments, a CNX-binding antibody clone is selected from:1D3, 1 D6, 1 E1, 1 E6, 2C6, 2H6, 3D1. 2G9, 2G12, 2H5, 3F8, 3F9, 4G9, 5A3, 5E8, C001, C008, C010, C023, C025, C040, C046 and C117.

[0225] In some embodiments, the antigen-binding moiety comprises a VH region according to one of (1 ) to (19) below:

[0226] (1 ) a VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:2

[0227] HC-CDR2 having the amino acid sequence of SEQ ID NO:3

[0228] HC-CDR3 having the amino acid sequence of SEQ ID NO:4, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0229] (2) a VH region incorporating the following CDRs:

[0230] HC-CDR1 having the amino acid sequence of SEQ ID NO:18

[0231] HC-CDR2 having the amino acid sequence of SEQ ID NO:19

[0232] HC-CDR3 having the amino acid sequence of SEQ ID NQ:20, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0233] (3) a VH region incorporating the following CDRs:

[0234] HC-CDR1 having the amino acid sequence of SEQ ID NO:33

[0235] HC-CDR2 having the amino acid sequence of SEQ ID NO:34

[0236] HC-CDR3 having the amino acid sequence of SEQ ID NO:35, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0237] (4) a VH region incorporating the following CDRs:

[0238] HC-CDR1 having the amino acid sequence of SEQ ID NO:48

[0239] HC-CDR2 having the amino acid sequence of SEQ ID NO:3

[0240] HC-CDR3 having the amino acid sequence of SEQ ID NO:49, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0241] (5) a VH region incorporating the following CDRs:

[0242] HC-CDR1 having the amino acid sequence of SEQ ID NO:61

[0243] HC-CDR2 having the amino acid sequence of SEQ ID NO:62

[0244] HC-CDR3 having the amino acid sequence of SEQ ID NO:63, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0245] (6) a VH region incorporating the following CDRs:

[0246] HC-CDR1 having the amino acid sequence of SEQ ID NO:2

[0247] HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:83, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0248] (7) a VH region incorporating the following CDRs:

[0249] HC-CDR1 having the amino acid sequence of SEQ ID NO:48

[0250] HC-CDR2 having the amino acid sequence of SEQ ID NO:3

[0251] HC-CDR3 having the amino acid sequence of SEQ ID NO:86, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0252] (8) a VH region incorporating the following CDRs:

[0253] HC-CDR1 having the amino acid sequence of SEQ ID NO:61

[0254] HC-CDR2 having the amino acid sequence of SEQ ID NO:95

[0255] HC-CDR3 having the amino acid sequence of SEQ ID NO:96, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0256] (9) a VH region incorporating the following CDRs:

[0257] HC-CDR1 having the amino acid sequence of SEQ ID NQ:108

[0258] HC-CDR2 having the amino acid sequence of SEQ ID NQ:109

[0259] HC-CDR3 having the amino acid sequence of SEQ ID NQ:110, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0260] (10) a VH region incorporating the following CDRs:

[0261] HC-CDR1 having the amino acid sequence of SEQ ID NO:2

[0262] HC-CDR2 having the amino acid sequence of SEQ ID NO:3

[0263] HC-CDR3 having the amino acid sequence of SEQ ID NO:122, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0264] (11 ) a VH region incorporating the following CDRs:

[0265] HC-CDR1 having the amino acid sequence of SEQ ID NO:132

[0266] HC-CDR2 having the amino acid sequence of SEQ ID NO:133

[0267] HC-CDR3 having the amino acid sequence of SEQ ID NO:134, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0268] (12) a VH region incorporating the following CDRs:

[0269] HC-CDR1 having the amino acid sequence of SEQ ID NO:146

[0270] HC-CDR2 having the amino acid sequence of SEQ ID NO:147

[0271] HC-CDR3 having the amino acid sequence of SEQ ID NO:148, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0272] (13) a VH region incorporating the following CDRs:

[0273] HC-CDR1 having the amino acid sequence of SEQ ID NO:48

[0274] HC-CDR2 having the amino acid sequence of SEQ ID NO:3

[0275] HC-CDR3 having the amino acid sequence of SEQ ID NO:156, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0276] (14) a VH region incorporating the following CDRs:

[0277] HC-CDR1 having the amino acid sequence of SEQ ID NO:166

[0278] HC-CDR2 having the amino acid sequence of SEQ ID NO:167

[0279] HC-CDR3 having the amino acid sequence of SEQ ID NO:168, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0280] (15) a VH region incorporating the following CDRs:

[0281] HC-CDR1 having the amino acid sequence of SEQ ID NO:185

[0282] HC-CDR2 having the amino acid sequence of SEQ ID NO:186

[0283] HC-CDR3 having the amino acid sequence of SEQ ID NO:187, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0284] (16) a VH region incorporating the following CDRs:

[0285] HC-CDR1 having the amino acid sequence of SEQ ID NO:48

[0286] HC-CDR2 having the amino acid sequence of SEQ ID NO:199

[0287] HC-CDR3 having the amino acid sequence of SEQ ID NQ:200, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0288] (17) a VH region incorporating the following CDRs:

[0289] HC-CDR1 having the amino acid sequence of SEQ ID NO:48

[0290] HC-CDR2 having the amino acid sequence of SEQ ID NO:211

[0291] HC-CDR3 having the amino acid sequence of SEQ ID NO:212, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0292] (18) a VH region incorporating the following CDRs:

[0293] HC-CDR1 having the amino acid sequence of SEQ ID NO:222

[0294] HC-CDR2 having the amino acid sequence of SEQ ID NO:223

[0295] HC-CDR3 having the amino acid sequence of SEQ ID NO:224, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0296] (19) a VH region incorporating the following CDRs:

[0297] HC-CDR1 having the amino acid sequence of SEQ ID NO:185

[0298] HC-CDR2 having the amino acid sequence of SEQ ID NO:243

[0299] HC-CDR3 having the amino acid sequence of SEQ ID NO:244, or a variant thereof in which 1 or 2 or 3 amino acids in HC-CDR1 , and / or in which 1 or 2 or 3 amino acids in HC-CDR2, and / or in which 1 or 2 or 3 amino acids in HC-CDR3 are substituted with another amino acid.

[0300] In some embodiments, the antigen-binding moiety comprises a VH region according to one of (20) to (37) below:

[0301] (20) a VH region incorporating the following FRs:

[0302] HC-FR1 having the amino acid sequence of SEQ ID NO:5

[0303] HC-FR2 having the amino acid sequence of SEQ ID NO:6

[0304] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0305] HC-FR4 having the amino acid sequence of SEQ ID NO:8, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0306] (21 ) a VH region incorporating the following FRs:

[0307] HC-FR1 having the amino acid sequence of SEQ ID NO:21

[0308] HC-FR2 having the amino acid sequence of SEQ ID NO:22 HC-FR3 having the amino acid sequence of SEQ ID NO:23

[0309] HC-FR4 having the amino acid sequence of SEQ ID NO:8, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0310] (22) a VH region incorporating the following FRs:

[0311] HC-FR1 having the amino acid sequence of SEQ ID NO:36

[0312] HC-FR2 having the amino acid sequence of SEQ ID NO:37

[0313] HC-FR3 having the amino acid sequence of SEQ ID NO:38

[0314] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0315] (23) a VH region incorporating the following FRs:

[0316] HC-FR1 having the amino acid sequence of SEQ ID NQ:50

[0317] HC-FR2 having the amino acid sequence of SEQ ID NO:6

[0318] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0319] HC-FR4 having the amino acid sequence of SEQ ID NO:51 , or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0320] (24) a VH region incorporating the following FRs:

[0321] HC-FR1 having the amino acid sequence of SEQ ID NO:64

[0322] HC-FR2 having the amino acid sequence of SEQ ID NO:65

[0323] HC-FR3 having the amino acid sequence of SEQ ID NO:66

[0324] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0325] (25) a VH region incorporating the following FRs:

[0326] HC-FR1 having the amino acid sequence of SEQ ID NO:84

[0327] HC-FR2 having the amino acid sequence of SEQ ID NO:6

[0328] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0329] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid. (26) a VH region incorporating the following FRs:

[0330] HC-FR1 having the amino acid sequence of SEQ ID NO:87

[0331] HC-FR2 having the amino acid sequence of SEQ ID NO:6

[0332] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0333] HC-FR4 having the amino acid sequence of SEQ ID NO:51 , or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0334] (27) a VH region incorporating the following FRs:

[0335] HC-FR1 having the amino acid sequence of SEQ ID NO:97

[0336] HC-FR2 having the amino acid sequence of SEQ ID NO:98

[0337] HC-FR3 having the amino acid sequence of SEQ ID NO:66

[0338] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0339] (28) a VH region incorporating the following FRs:

[0340] HC-FR1 having the amino acid sequence of SEQ ID NO:111

[0341] HC-FR2 having the amino acid sequence of SEQ ID NO:112

[0342] HC-FR3 having the amino acid sequence of SEQ ID NO:113

[0343] HC-FR4 having the amino acid sequence of SEQ ID NO:51 , or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0344] (29) a VH region incorporating the following FRs:

[0345] HC-FR1 having the amino acid sequence of SEQ ID NO:123

[0346] HC-FR2 having the amino acid sequence of SEQ ID NO:6

[0347] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0348] HC-FR4 having the amino acid sequence of SEQ ID NO:51 , or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0349] (30) a VH region incorporating the following FRs:

[0350] HC-FR1 having the amino acid sequence of SEQ ID NO:135

[0351] HC-FR2 having the amino acid sequence of SEQ ID NO:136

[0352] HC-FR3 having the amino acid sequence of SEQ ID NO:137

[0353] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0354] (31 ) a VH region incorporating the following FRs:

[0355] HC-FR1 having the amino acid sequence of SEQ ID NO:149

[0356] HC-FR2 having the amino acid sequence of SEQ ID NO:150

[0357] HC-FR3 having the amino acid sequence of SEQ ID NO:151

[0358] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0359] (32) a VH region incorporating the following FRs:

[0360] HC-FR1 having the amino acid sequence of SEQ ID NO:87

[0361] HC-FR2 having the amino acid sequence of SEQ ID NO:169

[0362] HC-FR3 having the amino acid sequence of SEQ ID NO:168

[0363] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0364] (33) a VH region incorporating the following FRs:

[0365] HC-FR1 having the amino acid sequence of SEQ ID NO:188

[0366] HC-FR2 having the amino acid sequence of SEQ ID NO:189

[0367] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0368] HC-FR4 having the amino acid sequence of SEQ ID NO:8, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0369] (34) a VH region incorporating the following FRs:

[0370] HC-FR1 having the amino acid sequence of SEQ ID NQ:201

[0371] HC-FR2 having the amino acid sequence of SEQ ID NQ:202

[0372] HC-FR3 having the amino acid sequence of SEQ ID NQ:203

[0373] HC-FR4 having the amino acid sequence of SEQ ID NO:8, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0374] (35) a VH region incorporating the following FRs:

[0375] HC-FR1 having the amino acid sequence of SEQ ID NO:213 HC-FR2 having the amino acid sequence of SEQ ID NO:214

[0376] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0377] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0378] (36) a VH region incorporating the following FRs:

[0379] HC-FR1 having the amino acid sequence of SEQ ID NO:225

[0380] HC-FR2 having the amino acid sequence of SEQ ID NO:226

[0381] HC-FR3 having the amino acid sequence of SEQ ID NO:227

[0382] HC-FR4 having the amino acid sequence of SEQ ID NO:8, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0383] (37) a VH region incorporating the following FRs:

[0384] HC-FR1 having the amino acid sequence of SEQ ID NO:245

[0385] HC-FR2 having the amino acid sequence of SEQ ID NO:246

[0386] HC-FR3 having the amino acid sequence of SEQ ID NQ:190

[0387] HC-FR4 having the amino acid sequence of SEQ ID NO:39, or a variant thereof in which 1 or 2 or 3 amino acids in HC-FR1 , and / or in which 1 or 2 or 3 amino acids in HC-FR2, and / or in which 1 or 2 or 3 amino acids in HC-FR3, and / or in which 1 or 2 or 3 amino acids in HC-FR4 are substituted with another amino acid.

[0388] In some embodiments, the antigen-binding moiety comprises a VH region comprising the CDRs according to any one of (1 ) to (19) above, and the FRs according to any one of (20) to (37) above.

[0389] In some embodiments, the antigen-binding moiety comprises a VH region according to one of (38) to (56) below:

[0390] (38) a VH region comprising the CDRs according to (1 ) and the FRs according to (20).

[0391] (39) a VH region comprising the CDRs according to (2) and the FRs according to (21 ).

[0392] (40) a VH region comprising the CDRs according to (3) and the FRs according to (22).

[0393] (41 ) a VH region comprising the CDRs according to (4) and the FRs according to (23).

[0394] (42) a VH region comprising the CDRs according to (5) and the FRs according to (24).

[0395] (43) a VH region comprising the CDRs according to (6) and the FRs according to (25). (44) a VH region comprising the CDRs according to (7) and the FRs according to (26).

[0396] (45) a VH region comprising the CDRs according to (8) and the FRs according to (27).

[0397] (46) a VH region comprising the CDRs according to (9) and the FRs according to (28).

[0398] (47) a VH region comprising the CDRs according to (10) and the FRs according to (29).

[0399] (48) a VH region comprising the CDRs according to (11 ) and the FRs according to (30).

[0400] (49) a VH region comprising the CDRs according to (12) and the FRs according to (31 ).

[0401] (50) a VH region comprising the CDRs according to (13) and the FRs according to (26).

[0402] (51 ) a VH region comprising the CDRs according to (14) and the FRs according to (32).

[0403] (52) a VH region comprising the CDRs according to (15) and the FRs according to (33).

[0404] (53) a VH region comprising the CDRs according to (16) and the FRs according to (34).

[0405] (54) a VH region comprising the CDRs according to (17) and the FRs according to (35).

[0406] (55) a VH region comprising the CDRs according to (18) and the FRs according to (36).

[0407] (56) a VH region comprising the CDRs according to (19) and the FRs according to (37).

[0408] In some embodiments, the antigen-binding moiety comprises a VH region according to one of (57) to (75) below:

[0409] (57) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:1.

[0410] (58) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:17.

[0411] (59) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:32. (60) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:47.

[0412] (61 ) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:60.

[0413] (62) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:82.

[0414] (63) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:85.

[0415] (64) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:94.

[0416] (65) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NQ:107.

[0417] (66) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:121 .

[0418] (67) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:131 .

[0419] (68) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:145.

[0420] (69) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:155. (70) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:165.

[0421] (71 ) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:184.

[0422] (72) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:198.

[0423] (73) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NQ:210.

[0424] (74) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:221 .

[0425] (75) a VH region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:242.

[0426] In some embodiments, the antigen-binding moiety comprises a VL region according to one of (76) to (97) below:

[0427] (76) a VL region incorporating the following CDRs:

[0428] LC-CDR1 having the amino acid sequence of SEQ ID NQ:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0429] (77) a VL region incorporating the following CDRs:

[0430] LC-CDR1 having the amino acid sequence of SEQ ID NO:25 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:27, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0431] (78) a VL region incorporating the following CDRs:

[0432] LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42

[0433] LC-CDR3 having the amino acid sequence of SEQ ID NO:43, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0434] (79) a VL region incorporating the following CDRs:

[0435] LC-CDR1 having the amino acid sequence of SEQ ID NO:53

[0436] LC-CDR2 having the amino acid sequence of SEQ ID NO:54

[0437] LC-CDR3 having the amino acid sequence of SEQ ID NO:55, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0438] (80) a VL region incorporating the following CDRs:

[0439] LC-CDR1 having the amino acid sequence of SEQ ID NO:68

[0440] LC-CDR2 having the amino acid sequence of SEQ ID NO:26

[0441] LC-CDR3 having the amino acid sequence of SEQ ID NO:69, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0442] (81 ) a VL region incorporating the following CDRs:

[0443] LC-CDR1 having the amino acid sequence of SEQ ID NO:73

[0444] LC-CDR2 having the amino acid sequence of SEQ ID NO:26

[0445] LC-CDR3 having the amino acid sequence of SEQ ID NO:74, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0446] (82) a VL region incorporating the following CDRs:

[0447] LC-CDR1 having the amino acid sequence of SEQ ID NO:78

[0448] LC-CDR2 having the amino acid sequence of SEQ ID NO:79

[0449] LC-CDR3 having the amino acid sequence of SEQ ID NQ:80, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0450] (83) a VL region incorporating the following CDRs:

[0451] LC-CDR1 having the amino acid sequence of SEQ ID NO:89

[0452] LC-CDR2 having the amino acid sequence of SEQ ID NO:11

[0453] LC-CDR3 having the amino acid sequence of SEQ ID NQ:90, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0454] (84) a VL region incorporating the following CDRs:

[0455] LC-CDR1 having the amino acid sequence of SEQ ID NQ:101 LC-CDR2 having the amino acid sequence of SEQ ID NQ:102

[0456] LC-CDR3 having the amino acid sequence of SEQ ID NO:103, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0457] (85) a VL region incorporating the following CDRs:

[0458] LC-CDR1 having the amino acid sequence of SEQ ID NO:115

[0459] LC-CDR2 having the amino acid sequence of SEQ ID NO:116

[0460] LC-CDR3 having the amino acid sequence of SEQ ID NO:117, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0461] (86) a VL region incorporating the following CDRs:

[0462] LC-CDR1 having the amino acid sequence of SEQ ID NO:125

[0463] LC-CDR2 having the amino acid sequence of SEQ ID NO:126

[0464] LC-CDR3 having the amino acid sequence of SEQ ID NO:127, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0465] (87) a VL region incorporating the following CDRs:

[0466] LC-CDR1 having the amino acid sequence of SEQ ID NO:139

[0467] LC-CDR2 having the amino acid sequence of SEQ ID NQ:140

[0468] LC-CDR3 having the amino acid sequence of SEQ ID NQ:80, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0469] (88) a VL region incorporating the following CDRs:

[0470] LC-CDR1 having the amino acid sequence of SEQ ID NO:41

[0471] LC-CDR2 having the amino acid sequence of SEQ ID NO:42

[0472] LC-CDR3 having the amino acid sequence of SEQ ID NO:153, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0473] (89) a VL region incorporating the following CDRs:

[0474] LC-CDR1 having the amino acid sequence of SEQ ID NO:158

[0475] LC-CDR2 having the amino acid sequence of SEQ ID NO:159

[0476] LC-CDR3 having the amino acid sequence of SEQ ID NQ:160, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0477] (90) a VL region incorporating the following CDRs:

[0478] LC-CDR1 having the amino acid sequence of SEQ ID NO:171 LC-CDR2 having the amino acid sequence of SEQ ID NO:172

[0479] LC-CDR3 having the amino acid sequence of SEQ ID NO:173, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0480] (91 ) a VL region incorporating the following CDRs:

[0481] LC-CDR1 having the amino acid sequence of SEQ ID NO:179

[0482] LC-CDR2 having the amino acid sequence of SEQ ID NQ:180

[0483] LC-CDR3 having the amino acid sequence of SEQ ID NO:173, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0484] (92) a VL region incorporating the following CDRs:

[0485] LC-CDR1 having the amino acid sequence of SEQ ID NO:73

[0486] LC-CDR2 having the amino acid sequence of SEQ ID NO:26

[0487] LC-CDR3 having the amino acid sequence of SEQ ID NO:194, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0488] (93) a VL region incorporating the following CDRs:

[0489] LC-CDR1 having the amino acid sequence of SEQ ID NQ:205

[0490] LC-CDR2 having the amino acid sequence of SEQ ID NO:42

[0491] LC-CDR3 having the amino acid sequence of SEQ ID NQ:206, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0492] (94) a VL region incorporating the following CDRs:

[0493] LC-CDR1 having the amino acid sequence of SEQ ID NO:216

[0494] LC-CDR2 having the amino acid sequence of SEQ ID NO:172

[0495] LC-CDR3 having the amino acid sequence of SEQ ID NO:217, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0496] (95) a VL region incorporating the following CDRs:

[0497] LC-CDR1 having the amino acid sequence of SEQ ID NO:229

[0498] LC-CDR2 having the amino acid sequence of SEQ ID NO:172

[0499] LC-CDR3 having the amino acid sequence of SEQ ID NQ:230, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0500] (96) a VL region incorporating the following CDRs:

[0501] LC-CDR1 having the amino acid sequence of SEQ ID NO:235 LC-CDR2 having the amino acid sequence of SEQ ID NO:236

[0502] LC-CDR3 having the amino acid sequence of SEQ ID NO:237, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0503] (97) a VL region incorporating the following CDRs:

[0504] LC-CDR1 having the amino acid sequence of SEQ ID NO:248

[0505] LC-CDR2 having the amino acid sequence of SEQ ID NO:249

[0506] LC-CDR3 having the amino acid sequence of SEQ ID NQ:250, or a variant thereof in which 1 or 2 or 3 amino acids in LC-CDR1 , and / or in which 1 or 2 or 3 amino acids in LC-CDR2, and / or in which 1 or 2 or 3 amino acids in LC-CDR3 are substituted with another amino acid.

[0507] In some embodiments, the antigen-binding moiety comprises a VL region according to one of (98) to

[0508] (119) below:

[0509] (98) a VL region incorporating the following FRs:

[0510] LC-FR1 having the amino acid sequence of SEQ ID NO:13

[0511] LC-FR2 having the amino acid sequence of SEQ ID NO:14

[0512] LC-FR3 having the amino acid sequence of SEQ ID NO:15

[0513] LC-FR4 having the amino acid sequence of SEQ ID NO:16, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0514] (99) a VL region incorporating the following FRs:

[0515] LC-FR1 having the amino acid sequence of SEQ ID NO:28

[0516] LC-FR2 having the amino acid sequence of SEQ ID NO:29

[0517] LC-FR3 having the amino acid sequence of SEQ ID NQ:30

[0518] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0519] (100) a VL region incorporating the following FRs:

[0520] LC-FR1 having the amino acid sequence of SEQ ID NO:44

[0521] LC-FR2 having the amino acid sequence of SEQ ID NO:45

[0522] LC-FR3 having the amino acid sequence of SEQ ID NO:46

[0523] LC-FR4 having the amino acid sequence of SEQ ID NO:16, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0524] (101) a VL region incorporating the following FRs: LC-FR1 having the amino acid sequence of SEQ ID NO:56

[0525] LC-FR2 having the amino acid sequence of SEQ ID NO:57

[0526] LC-FR3 having the amino acid sequence of SEQ ID NO:58

[0527] LC-FR4 having the amino acid sequence of SEQ ID NO:59, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0528] (102) a VL region incorporating the following FRs:

[0529] LC-FR1 having the amino acid sequence of SEQ ID NO:28

[0530] LC-FR2 having the amino acid sequence of SEQ ID NQ:70

[0531] LC-FR3 having the amino acid sequence of SEQ ID NO:71

[0532] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0533] (103) a VL region incorporating the following FRs:

[0534] LC-FR1 having the amino acid sequence of SEQ ID NO:28

[0535] LC-FR2 having the amino acid sequence of SEQ ID NO:75

[0536] LC-FR3 having the amino acid sequence of SEQ ID NO:76

[0537] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0538] (104) a VL region incorporating the following FRs:

[0539] LC-FR1 having the amino acid sequence of SEQ ID NO:28

[0540] LC-FR2 having the amino acid sequence of SEQ ID NO:81

[0541] LC-FR3 having the amino acid sequence of SEQ ID NO:76

[0542] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0543] (105) a VL region incorporating the following FRs:

[0544] LC-FR1 having the amino acid sequence of SEQ ID NO:91

[0545] LC-FR2 having the amino acid sequence of SEQ ID NO:92

[0546] LC-FR3 having the amino acid sequence of SEQ ID NO:93

[0547] LC-FR4 having the amino acid sequence of SEQ ID NO:16, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0548] (106) a VL region incorporating the following FRs:

[0549] LC-FR1 having the amino acid sequence of SEQ ID NO:104

[0550] LC-FR2 having the amino acid sequence of SEQ ID NO:105

[0551] LC-FR3 having the amino acid sequence of SEQ ID NQ:106

[0552] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0553] (107) a VL region incorporating the following FRs:

[0554] LC-FR1 having the amino acid sequence of SEQ ID NO:118

[0555] LC-FR2 having the amino acid sequence of SEQ ID NO:119

[0556] LC-FR3 having the amino acid sequence of SEQ ID NQ:120

[0557] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0558] (108) a VL region incorporating the following FRs:

[0559] LC-FR1 having the amino acid sequence of SEQ ID NO:128

[0560] LC-FR2 having the amino acid sequence of SEQ ID NO:129

[0561] LC-FR3 having the amino acid sequence of SEQ ID NQ:130

[0562] LC-FR4 having the amino acid sequence of SEQ ID NO:16, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0563] (109) a VL region incorporating the following FRs:

[0564] LC-FR1 having the amino acid sequence of SEQ ID NO:141

[0565] LC-FR2 having the amino acid sequence of SEQ ID NO:142

[0566] LC-FR3 having the amino acid sequence of SEQ ID NO:143

[0567] LC-FR4 having the amino acid sequence of SEQ ID NO:144, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0568] (110) a VL region incorporating the following FRs:

[0569] LC-FR1 having the amino acid sequence of SEQ ID NO:44 LC-FR2 having the amino acid sequence of SEQ ID NO:45

[0570] LC-FR3 having the amino acid sequence of SEQ ID NO:46

[0571] LC-FR4 having the amino acid sequence of SEQ ID NO:154, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0572] (111 ) a VL region incorporating the following FRs:

[0573] LC-FR1 having the amino acid sequence of SEQ ID NO:161

[0574] LC-FR2 having the amino acid sequence of SEQ ID NO:162

[0575] LC-FR3 having the amino acid sequence of SEQ ID NO:163

[0576] LC-FR4 having the amino acid sequence of SEQ ID NO:164, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0577] (112) a VL region incorporating the following FRs:

[0578] LC-FR1 having the amino acid sequence of SEQ ID NO:174

[0579] LC-FR2 having the amino acid sequence of SEQ ID NO:175

[0580] LC-FR3 having the amino acid sequence of SEQ ID NO:176

[0581] LC-FR4 having the amino acid sequence of SEQ ID NO:177, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0582] (113) a VL region incorporating the following FRs:

[0583] LC-FR1 having the amino acid sequence of SEQ ID NO:181

[0584] LC-FR2 having the amino acid sequence of SEQ ID NO:182

[0585] LC-FR3 having the amino acid sequence of SEQ ID NO:183

[0586] LC-FR4 having the amino acid sequence of SEQ ID NO:177, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0587] (114) a VL region incorporating the following FRs:

[0588] LC-FR1 having the amino acid sequence of SEQ ID NO:28

[0589] LC-FR2 having the amino acid sequence of SEQ ID NO:75

[0590] LC-FR3 having the amino acid sequence of SEQ ID NO:197

[0591] LC-FR4 having the amino acid sequence of SEQ ID NO:177, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid. (115) a VL region incorporating the following FRs:

[0592] LC-FR1 having the amino acid sequence of SEQ ID NO:44

[0593] LC-FR2 having the amino acid sequence of SEQ ID NQ:207

[0594] LC-FR3 having the amino acid sequence of SEQ ID NQ:208

[0595] LC-FR4 having the amino acid sequence of SEQ ID NQ:209, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0596] (116) a VL region incorporating the following FRs:

[0597] LC-FR1 having the amino acid sequence of SEQ ID NO:218

[0598] LC-FR2 having the amino acid sequence of SEQ ID NO:219

[0599] LC-FR3 having the amino acid sequence of SEQ ID NQ:220

[0600] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0601] (117) a VL region incorporating the following FRs:

[0602] LC-FR1 having the amino acid sequence of SEQ ID NO:231

[0603] LC-FR2 having the amino acid sequence of SEQ ID NO:232

[0604] LC-FR3 having the amino acid sequence of SEQ ID NO:233

[0605] LC-FR4 having the amino acid sequence of SEQ ID NO:31 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0606] (118) a VL region incorporating the following FRs:

[0607] LC-FR1 having the amino acid sequence of SEQ ID NO:238

[0608] LC-FR2 having the amino acid sequence of SEQ ID NO:239

[0609] LC-FR3 having the amino acid sequence of SEQ ID NQ:240

[0610] LC-FR4 having the amino acid sequence of SEQ ID NO:241 , or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0611] (119) a VL region incorporating the following FRs:

[0612] LC-FR1 having the amino acid sequence of SEQ ID NO:231

[0613] LC-FR2 having the amino acid sequence of SEQ ID NO:251

[0614] LC-FR3 having the amino acid sequence of SEQ ID NO:252

[0615] LC-FR4 having the amino acid sequence of SEQ ID NO:253, or a variant thereof in which 1 or 2 or 3 amino acids in LC-FR1 , and / or in which 1 or 2 or 3 amino acids in LC-FR2, and / or in which 1 or 2 or 3 amino acids in LC-FR3, and / or in which 1 or 2 or 3 amino acids in LC-FR4 are substituted with another amino acid.

[0616] In some embodiments, the antigen-binding moiety comprises a VL region comprising the CDRs according to any one of (76) to (97) above, and the FRs according to any one of (98) to (119) above.

[0617] In some embodiments, the antigen-binding moiety comprises a VL region according to one of (120) to (142) below:

[0618] (120) a VL region comprising the CDRs according to (76) and the FRs according to (98).

[0619] (121) a VL region comprising the CDRs according to (77) and the FRs according to (99).

[0620] (123) a VL region comprising the CDRs according to (78) and the FRs according to (100).

[0621] (124) a VL region comprising the CDRs according to (79) and the FRs according to (101).

[0622] (125) a VL region comprising the CDRs according to (80) and the FRs according to (102).

[0623] (126) a VL region comprising the CDRs according to (81 ) and the FRs according to (103).

[0624] (127) a VL region comprising the CDRs according to (82) and the FRs according to (104).

[0625] (128) a VL region comprising the CDRs according to (83) and the FRs according to (105).

[0626] (129) a VL region comprising the CDRs according to (84) and the FRs according to (106).

[0627] (130) a VL region comprising the CDRs according to (85) and the FRs according to (107).

[0628] (131) a VL region comprising the CDRs according to (86) and the FRs according to (108).

[0629] (132) a VL region comprising the CDRs according to (87) and the FRs according to (109).

[0630] (133) a VL region comprising the CDRs according to (88) and the FRs according to (110).

[0631] (134) a VL region comprising the CDRs according to (89) and the FRs according to (111).

[0632] (135) a VL region comprising the CDRs according to (90) and the FRs according to (112).

[0633] (136) a VL region comprising the CDRs according to (91 ) and the FRs according to (113).

[0634] (137) a VL region comprising the CDRs according to (92) and the FRs according to (114). (138) a VL region comprising the CDRs according to (93) and the FRs according to (115).

[0635] (139) a VL region comprising the CDRs according to (94) and the FRs according to (116).

[0636] (140) a VL region comprising the CDRs according to (95) and the FRs according to (117).

[0637] (141) a VL region comprising the CDRs according to (96) and the FRs according to (118).

[0638] (142) a VL region comprising the CDRs according to (97) and the FRs according to (119).

[0639] In some embodiments, the antigen-binding moiety comprises a VL region according to one of (143) to (164) below:

[0640] (143) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:9.

[0641] (144) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:24.

[0642] (145) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NQ:40.

[0643] (146) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:52.

[0644] (147) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:67.

[0645] (148) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:72.

[0646] (149) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:77. (150) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:88.

[0647] (151) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NQ:100.

[0648] (152) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:114.

[0649] (153) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:124.

[0650] (154) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:138.

[0651] (155) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:152.

[0652] (156) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:157.

[0653] (157) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NQ:170.

[0654] (158) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:178.

[0655] (159) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:191 . (160) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:204.

[0656] (161) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:215.

[0657] (162) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:228.

[0658] (163) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:234.

[0659] (164) a VL region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:247.

[0660] In some embodiments, the antigen-binding moiety comprises a VH region according to any one of (1) to (75) above, and a VL region according to any one of (76) to (164) above.

[0661] In some embodiments described herein, one or more amino acids of an amino acid sequence referred to herein (e.g. an amino acid sequence of an antigen-binding moiety, e.g. an amino acid sequence of a CDR or VH / VL region) are substituted with another amino acid. A substitution comprises substitution of an amino acid residue with a non-identical 'replacement' amino acid residue. A replacement amino acid residue of a substitution according to the present disclosure may be a naturally-occurring amino acid residue ( / .e. encoded by the genetic code) which is non-identical to the amino acid residue at the relevant position of the equivalent, unsubstituted amino acid sequence, selected from: alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (lie): leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Vai). In some embodiments, a replacement amino acid may be a non-naturally occurring amino acid residue - i.e. an amino acid residue other than those recited in the preceding sentence. Examples of non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine, aib, and other amino acid residue analogues such as those described in Ellman, et al., Meth. Enzym. 202 (1991 ) 301-336.

[0662] In some embodiments, a substitution may be biochemically conservative. In some embodiments, where an amino acid to be substituted is provided in one of rows 1 to 5 of the table below, the replacement amino acid of the substitution is another, non-identical amino acid provided in the same row:

[0663] By way of illustration, in some embodiments wherein substitution is of a Met residue, the replacement amino acid may be selected from Ala, Vai, Leu, lie, Trp, Tyr, Phe and Norleucine.

[0664] In some embodiments, a replacement amino acid in a substitution may have the same side chain polarity as the amino acid residue it replaces. In some embodiments, a replacement amino acid in a substitution may have the same side chain charge (at pH 7.4) as the amino acid residue it replaces:

[0665] That is, in some embodiments, a nonpolar amino acid is substituted with another, non-identical nonpolar amino acid. In some embodiments, a polar amino acid is substituted with another, non-identical polar amino acid. In some embodiments, an acidic polar amino acid is substituted with another, non-identical acidic polar amino acid. In some embodiments, a basic polar amino acid is substituted with another, non- identical basic polar amino acid. In some embodiments, a neutral amino acid is substituted with another, non-identical neutral amino acid. In some embodiments, a positive amino acid is substituted with another, non-identical positive amino acid. In some embodiments, a negative amino acid is substituted with another, non-identical negative amino acid.

[0666] In some embodiments, substitution(s) may be functionally conservative. That is, in some embodiments, the substitution may not affect (or may not substantially affect) one or more functional properties (e.g. target binding) of the antigen-binding molecule comprising the substitution as compared to the equivalent unsubstituted molecule.

[0667] The VH and VL region of an antigen-binding region of an antibody together constitute the Fv region. In some embodiments, the antigen-binding moiety according to the present disclosure comprises, or consists of, an Fv region that binds to CNX. In some embodiments, the VH and VL regions of the Fv are provided as single polypeptide joined by a linker region, i.e. a single chain Fv (scFv).

[0668] The VL and light chain constant (CL) region, and the VH region and heavy chain constant 1 (CH1 ) region of an antigen-binding region of an antibody together constitute the Fab region. In some embodiments, the antigen-binding moiety comprises a Fab region comprising a VH, a CH1, a VL and a CL (e.g. CK or CA). In some embodiments, the Fab region comprises a polypeptide comprising a VH and a CH1 (e.g. a VH- CH1 fusion polypeptide), and a polypeptide comprising a VL and a CL (e.g. a VL-CL fusion polypeptide). In some embodiments, the Fab region comprises a polypeptide comprising a VH and a CL (e.g. a VH-CL fusion polypeptide) and a polypeptide comprising a VL and a CH (e.g. a VL-CH1 fusion polypeptide); that is, in some embodiments, the Fab region is a CrossFab region. In some embodiments, the VH, CH1, VL and CL regions of the Fab or CrossFab are provided as single polypeptide joined by linker regions, i.e. as a single chain Fab (scFab) or a single chain CrossFab (scCrossFab).

[0669] In some embodiments, the antigen-binding moiety described herein comprises, or consists of, a whole antibody that binds to CNX. As used herein, ‘whole antibody’ refers to an antibody having a structure which is substantially similar to the structure of an immunoglobulin (Ig). Different kinds of immunoglobulins and their structures are described e.g. in Schroeder and Cavacini J Allergy Clin Immunol. (2010) 125(202): S41-S52, which is hereby incorporated by reference in its entirety.

[0670] Immunoglobulins of type G (i.e. IgG) are -150 kDa glycoproteins comprising two heavy chains and two light chains. From N- to C-terminus, the heavy chains comprise a VH followed by a heavy chain constant region comprising three constant domains (CH1, CH2, and CH3), and similarly the light chains comprise a VL followed by a CL. Depending on the heavy chain, immunoglobulins may be classed as IgG (e.g. lgG1 , lgG2, lgG3, lgG4), IgA (e.g. lgA1 , lgA2), IgD, IgE, or IgM. The light chain may be kappa (K) or lambda (A).

[0671] In some embodiments, the antigen-binding moiety comprises, or consists of, an IgG (e.g. lgG1, lgG2, lgG3, lgG4), IgA (e.g. lgA1, lgA2), IgD, IgE, or IgM that binds to CNX.

[0672] In some embodiments, the antigen-binding moiety of the present disclosure comprises one or more regions (e.g. CH1 , CH2, CH3, etc.) of an immunoglobulin heavy chain constant sequence. In some embodiments, the immunoglobulin heavy chain constant sequence is, or is derived from, the heavy chain constant sequence of an IgG (e.g. IgG 1 , lgG2, lgG3, lgG4), IgA (e.g. lgA1 , Ig A2), IgD, IgE or IgM, e.g. a human IgG (e.g. hlgG1 , hlgG2, hlgG3, hlgG4), hlgA (e.g. hlgA1, hlgA2), hlgD, hlgE or hlgM. In some embodiments, the immunoglobulin heavy chain constant sequence is, or is derived from, the heavy chain constant sequence of a human IgG 1 allotype (e.g. G1 m1, G1 m2, G1 m3 or G1 m17). In some embodiments, the antigen-binding moiety comprises an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:254, 259, 260 or 263.

[0673] In some embodiments, the antigen-binding moiety comprises a CH1 region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:255 or 261 . In some embodiments, the antigen-binding moiety comprises a CH2 region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:257. In some embodiments, the antigen-binding moiety comprises a CH3 region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:258 or 262.

[0674] In some embodiments, the antigen-binding moiety comprises a hinge region comprising an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:256, 264, 265 or 266.

[0675] It will be appreciated that CH2 and / or CH3 regions may be provided with further substitutions in accordance with modification to an Fc region of the antigen-binding moiety as described herein.

[0676] In some embodiments, the antigen-binding moiety of the present disclosure comprises one or more regions of an immunoglobulin light chain constant sequence. In some embodiments, the immunoglobulin light chain constant sequence is human immunoglobulin kappa constant (IGKC; CK). In some embodiments, the immunoglobulin light chain constant sequence is a human immunoglobulin lambda constant (IGLC; CA), e.g. IGLC1 , IGLC2, IGLC3, IGLC6 or IGLC7.

[0677] In some embodiments, the antigen-binding moiety comprises an amino acid sequence having at least 70% sequence identity more preferably one of at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, sequence identity to the amino acid sequence of SEQ ID NO:267, 268, 269, 270, 271 or 272.

[0678] In some embodiments, the antigen-binding moiety is or comprises a monoclonal antibody, or an antigenbinding fragment thereof.

[0679] In some embodiments, the antigen-binding moiety is or comprises a fully human antibody / antibody fragment. A fully human antibody / antibody fragment may be encoded by human nucleic acid sequence(s). A fully human antibody / antibody fragment may be devoid of non-human amino acid sequences. Commonly employed techniques for the production of fully human antibodies include (i) phage display, in which human antibody genes are expressed in phage display libraries, and (ii) production of antibodies in transgenic mice engineered to have human antibody genes (described in Park and Smolen, Advances in Protein Chemistry (2001) 56: 369-421). Briefly, in the human antibody genephage display technique, genes encoding the VH and VL chains are generated by PCR amplification and cloning from ‘naive’ human lymphocytes, and assembled into a library from which they can be expressed either as d isulfide-li n ked Fab fragments or as single-chain Fv (scFv) fragments. The Fab- or scFv- encoding genes are fused to a surface coat protein of filamentous bacteriophage and Fab or scFv capable of binding to the target of interest can then be identified by screening the library with antigen. Molecular evolution or affinity maturation procedures can be employed to enhance the affinity of the Fab / scFv fragment. In the transgenic mouse technique, mice in which the endogenous murine Ig gene loci have been replaced by homologous recombination with their human homologues are immunized with antigen, and monoclonal antibody is prepared by conventional hybridoma technology, to yield a fully human monoclonal antibody.

[0680] In some embodiments, the antigen-binding moiety of the present disclosure is a mouse antibody / antibody fragment. In some embodiments, the antibody / antibody fragment is obtained from phage display using a human naive antibody gene library.

[0681] In some embodiments, the antigen-binding moiety is a mouse / human chimeric antibody / antibody fragment ( / .e. an antigen-binding moiety comprising mouse antibody variable domains and human antibody constant regions). In some embodiments, the antigen-binding moiety is a humanised antibody / antibody fragment. In some embodiments, the antigen-binding moiety comprises mouse antibody CDRs and human antibody framework and constant regions.

[0682] Mouse / human chimeric antigen-binding moieties can be prepared from mouse antibodies by the process of chimerisation, e.g. as described in Human Monoclonal Antibodies: Methods and Protocols, Michael Steinitz (Editor), Methods in Molecular Biology 1060, Springer Protocols, Humana Press (2014), in Chapter 8 thereof, in particular section 3 of Chapter 8.

[0683] Humanised antigen-binding moieties can be prepared from mouse antibodies by the process of humanisation, e.g. as described in Human Monoclonal Antibodies: Methods and Protocols, Michael Steinitz (Editor), Methods in Molecular Biology 1060, Springer Protocols, Humana Press (2014), in Chapter 7 thereof, in particular section 3.1 of Chapter 7 entitled ‘Antibody Humanization’. Techniques for antibody humanisation are also described e.g. in Safdari et al., Biotechnol Genet Eng Rev (2013) 29:175- 86.

[0684] Aspects and embodiments of the present disclosure contemplate multispecific antigen-binding moieties. By ‘multispecific’ it is meant that the antigen-binding moiety displays specific binding to more than one target. In some embodiments, the antigen-binding moiety is a bispecific antigen-binding moiety. In some embodiments, the antigen-binding moiety comprises at least two different antigen-binding domains ( / .e. at least two antigen-binding domains, e.g. comprising non-identical VHs and VLs).

[0685] In some embodiments, the antigen-binding moiety binds to CNX and another target (e.g. an antigen other than CNX), and so is at least bispecific. The term ‘bispecific’ means that the antigen-binding moiety is able to bind specifically to at least two distinct antigenic determinants.

[0686] It will be appreciated that an antigen-binding moiety according to the present disclosure (e.g. a multispecific antigen-binding moiety) may comprise antigen-binding moieties capable of binding to the targets for which the antigen-binding moiety is specific. For example, an antigen-binding moiety that binds to CNX and an antigen other than CNX may comprise: (i) an antigen-binding moiety that binds to CNX, and (ii) an antigen-binding moiety that binds to an antigen other than CNX.

[0687] It will also be appreciated that an antigen-binding moiety according to the present disclosure (e.g. a multispecific antigen-binding moiety) may comprise antigen-binding polypeptides or antigen-binding polypeptide complexes capable of binding to the targets for which the antigen-binding moiety is specific.

[0688] In some embodiments, a component antigen-binding moiety of a larger antigen-binding moiety (e.g. a multispecific antigen-binding moiety) may be referred to e.g. as an ‘antigen-binding domain’ or ‘antigenbinding region’ of the larger antigen-binding moiety.

[0689] Multispecific antigen-binding moieties according to the present disclosure may be provided in any suitable format, such as those formats described in described in Brinkmann and Kontermann, MAbs (2017) 9(2): 182-212, which is hereby incorporated by reference in its entirety. Suitable formats include those shown in Figure 2 of Brinkmann and Kontermann, MAbs (2017) 9(2): 182-212: antibody conjugates, e.g. lgG2, F(ab’)2 or CovX-Body; IgG or IgG-like molecules, e.g. IgG, chimeric IgG, KX-body common HC; CH1 / CL fusion proteins, e.g. scFv2-CH1 / CL, VHH2-CH1 / CL; ‘variable domain only’ bispecific antigen-binding molecules, e.g. tandem scFv (taFV), triplebodies, diabodies (Db), dsDb, Db(kih), DART, scDB, dsFv- dsFv, tandAbs, triple heads, tandem dAbA / HH, tertravalent dAb.VHH; Non-lg fusion proteins, e.g. scFv2- albumin, scDb-albumin, taFv-albumin, taFv-toxin, miniantibody, DNL-Fab2, DNL-Fab2-scFv, DNL-Fab2- lgG-cytokine2, ImmTAC (TCR-scFv); modified Fc and CH3 fusion proteins, e.g. scFv-Fc(kih), scFv- Fc(CH3 charge pairs), scFv-Fc (EW-RVT), scFv-fc (HA-TF), scFv-Fc (SEEDbody), taFv-Fc(kih), scFv- Fc(kih)-Fv, Fab-Fc(kih)-scFv, Fab-scFv-Fc(kih), Fab-scFv-Fc(BEAT), Fab-scFv-Fc (SEEDbody), DART- Fc, scFv-CH3(kih), TriFabs; Fc fusions, e.g. Di-diabody, scDb-Fc, taFv-Fc, scFv-Fc-scFv, HCAb-VHH, Fab-scFv-Fc, scFv4-lg, scFv2-Fcab; CH3 fusions, e.g. Dia-diabody, scDb-CH3; IgE / IgM CH2 fusions, e.g. scFv-EHD2-scFv, scFvMHD2-scFv; Fab fusion proteins, e.g. Fab-scFv (bibody), Fab-scFv2 (tribody), Fab- Fv, Fab-dsFv, Fab-VHH, orthogonal Fab-Fab; non-lg fusion proteins, e.g. DNL-Fabs, DNL-Fab2-scFv, DNL-Fab2-lgG-cytokine2; asymmetric IgG or IgG-like molecules, e.g. IgG(kih), IgG(kih) common LC, ZW1 IgG common LC, Biclonics common LC, CrossMab, CrossMab(kih), scFab-lgG(kih), Fab-scFab-lgG(kih), orthogonal Fab IgG(kih), DuetMab, CH3 charge pairs + CH1 / CL charge pairs, hinge / CH3 charge pairs, SEED-body, Duobody, four-in-one-CrossMab(kih), LUZ-Y common LC; LUZ-Y scFab-IgG, FcFc*; appended and Fc-modified IgGs, e.g. lgG(kih)-Fv, IgG HA-TF-Fv, lgG(kih)scFab, scFab-Fc(kih)-scFv2, scFab-Fc(kih)-scFv, half DVD-lg, DVI-lg (four-in-one), CrossMab-Fab; modified Fc and CH3 fusion proteins, e.g. Fab-Fc(kih)-scFv, Fab-scFv-Fc(kih), Fab-scFv-Fc(BEAT), Fab-scFv-Fc-SEEDbody, TriFab; appended IgGs - HC fusions, e.g. IgG-HC, scFv, IgG-dAb, IgG-taFV, IgG-CrossFab, IgG-orthogonal Fab, IgG-(CaCp) Fab, scFv-HC-IgG, tandem Fab-IgG (orthogonal Fab), Fab-lgG(CaCp Fab), Fab-lgG(CR3), Fab-hinge-lgG(CR3); appended IgGs - LC fusions, e.g. IgG-scFv(LC), scFv(LC)-lgG, dAb-IgG; appended IgGs - HC and LC fusions, e.g. DVD-lg, TVD-lg, CODV-lg, scFv4-lgG, Zybody; Fc fusions, e.g. Fab-scFv- Fc, scFv4-lg; F(ab’)2 fusions, e.g. F(ab’)2-scFv2; CH1 / CL fusion proteins e.g. scFv2-CH1-hinge / CL; modified IgGs, e.g. DAF (two-in one-IgG), DutaMab, Mab2; and non-lg fusions, e.g. DNL-Fab4-lgG.

[0690] The skilled person is able to design and prepare multispecific antigen-binding moieties. Methods for producing multispecific antigen-binding moieties include chemically crosslinking antibodies or antibody fragments, e.g. with reducible disulphide or non-reducible thioether bonds, for example as described in Segal and Bast, 2001. Production of Bispecific Antigen-binding molecules. Current Protocols in Immunology. 14: IV:2.13:2.13.1—2.13.16, which is hereby incorporated by reference in its entirety. For example, A / -succinimidyl-3-(-2-pyridyldithio)-propionate (SPDP) can be used to chemically crosslink e.g. Fab fragments via hinge region SH- groups, to create disulfide-linked bispecific F(ab)2 heterodimers.

[0691] Other methods for producing multispecific antigen-binding moieties include fusing antibody-producing hybridomas e.g. with polyethylene glycol, to produce a quadroma cell capable of secreting bispecific antibody, for example as described in D. M. and Bast, B. J. 2001 . Production of Bispecific Antigen-binding molecules. Current Protocols in Immunology. 14: IV:2.13:2.13.1 — 2.13.16.

[0692] Multispecific antigen-binding moieties according to the present disclosure can also be produced recombinantly, by expression from e.g. a nucleic acid construct encoding polypeptides for the antigenbinding moieties / domains / regions, for example as described in Antibody Engineering: Methods and Protocols, Second Edition (Humana Press, 2012), at Chapter 40: Production of Bispecific Antigen-binding molecules: Diabodies and Tandem scFv (Hornig and Farber-Schwarz), or French, How to make bispecific antigen-binding molecules, Methods Mol. Med. 2000; 40:333-339, the entire contents of both of which are hereby incorporated by reference.

[0693] For example, a DNA construct encoding the light and heavy chain variable domains for the two antigenbinding fragments ( / .e. the light and heavy chain variable domains for the antigen-binding fragment capable of binding CNX, and the light and heavy chain variable domains for the antigen-binding fragment capable of binding to another target protein), and including sequences encoding a suitable linker or dimerization domain between the antigen-binding fragments can be prepared by molecular cloning techniques. Recombinant bispecific antibody can thereafter be produced by expression (e.g. in vitro) of the construct in a suitable host cell (e.g. a mammalian host cell), and expressed recombinant bispecific antibody can then optionally be purified.

[0694] Fc regions

[0695] In some embodiments, the antigen-binding molecules of the present disclosure (e.g. an antigen-binding moiety thereof) comprise an Fc region. An Fc region is composed of CH2 and CH3 regions from one polypeptide, and CH2 and CH3 regions from another polypeptide. The CH2 and CH3 regions from the two polypeptides together form the Fc region.

[0696] Fc-mediated functions include Fc receptor binding, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), formation of the membrane attack complex (MAC), cell degranulation, cytokine and / or chemokine production, and antigen processing and presentation. Modifications to antibody Fc regions that influence Fc-mediated functions are known in the art, such as those described e.g. in Wang et al., Protein Cell (2018) 9(1):63-73, which is hereby incorporated by reference in its entirety. Exemplary Fc region modifications known to influence antibody effector function are summarised in Table 1 of Wang et al., Protein Cell (2018) 9(1 ):63-73. In some embodiments, the antigen-binding molecule of the present disclosure (e.g. an antigen-binding moiety thereof) comprises an Fc region comprising modification to increase or reduce an Fc-mediated function as compared to an antigen-binding molecule comprising the corresponding unmodified Fc region.

[0697] Where an Fc region / CH2 / CH3 is described as comprising modification(s) ‘corresponding to’ reference substitution(s), equivalent substitution(s) in the homologous Fc / CH2 / CH3 are contemplated. By way of illustration, L234A / L235A substitutions in human lgG1 (numbered according to the EU numbering system as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991) correspond to L to A substitutions at positions 117 and 118 of the mouse Ig gamma-2A chain C region (UniProtKB: P01863-1 , v1 ).

[0698] Where an Fc region is described as comprising a modification, the modification may be present in one or both of the polypeptide chains which together form the Fc region.

[0699] In some embodiments, the antigen-binding molecule of the present disclosure comprises an Fc region comprising modification. In some embodiments, the antigen-binding molecule of the present disclosure comprises an Fc region comprising modification in one or more of the CH2 and / or CH3 regions.

[0700] In some embodiments, the Fc region comprises modification to increase an Fc-mediated function. In some embodiments, the Fc region comprises modification to increase ADCC. In some embodiments, the Fc region comprises modification to increase ADCP. In some embodiments, the Fc region comprises modification to increase CDC. An antigen-binding molecule comprising an Fc region comprising modification to increase an Fc-mediated function (e.g. ADCC, ADCP, CDC) induces an increased level of the relevant effector function as compared to an antigen-binding molecule comprising the corresponding unmodified Fc region.

[0701] In some embodiments, the Fc region comprises modification to increase binding to an Fc receptor. In some embodiments, the Fc region comprises modification to increase binding to an Fey receptor. In some embodiments, the Fc region comprises modification to increase binding to one or more of FcyRI, FcyRlla, FcyRllb, FcyRllc, FcyRllla and FcyRlllb. In some embodiments, the Fc region comprises modification to increase binding to FcyRllla. In some embodiments, the Fc region comprises modification to increase binding to FcyRlla. In some embodiments, the Fc region comprises modification to increase binding to FcyRllb. In some embodiments, the Fc region comprises modification to increase binding to FcRn. In some embodiments, the Fc region comprises modification to increase binding to a complement protein. In some embodiments, the Fc region comprises modification to increase binding to C1q. In some embodiments, the Fc region comprises modification to promote hexamerisation of the antigen-binding molecule. In some embodiments, the Fc region comprises modification to increase antigen-binding molecule half-life. In some embodiments, the Fc region comprises modification to increase coengagement.

[0702] In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions F243L / R292P / Y300L / V305I / P396L as described in Stavenhagen et al. Cancer Res. (2007) 67:8882-8890. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions S239D / I332E or S239D / I332E / A330L as described in Lazar et al., Proc Natl Acad Sci USA. (2006)103:4005-4010. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions S298A / E333A / K334A as described in Shields et al., J Biol Chem. (2001) 276:6591-6604. In some embodiments, the Fc region comprises modification to one of heavy chain polypeptides corresponding to the combination of substitutions L234Y / L235Q / G236W / S239M / H268D / D270E / S298A, and modification to the other heavy chain polypeptide corresponding to the combination of substitutions D270E / K326D / A330M / K334E, as described in Mimoto et al., MAbs. (2013): 5:229-236. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions G236A / S239D / I332E as described in Richards et al., Mol Cancer Ther. (2008) 7:2517-2527.

[0703] In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions K326W / E333S as described in Idusogie et al. J Immunol. (2001 ) 166(4):2571-5. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions S267E / H268F / S324T as described in Moore et al. MAbs. (2010) 2(2):181-9. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions described in Natsume et al., Cancer Res. (2008) 68(10):3863-72. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions E345R / E430G / S440Y as described in Diebolder et al. Science (2014) 343(6176): 1260-3.

[0704] In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions M252Y / S254T / T256E as described in Dall’Acqua et al. J Immunol. (2002) 169:5171-5180. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions M428L / N434S as described in Zalevsky et al. Nat Biotechnol. (2010) 28:157-159.

[0705] In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions S267E / L328F as described in Chu et al., Mol Immunol. (2008) 45:3926-3933. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions N325S / L328F as described in Shang et al. Biol Chem. (2014) 289:15309-15318.

[0706] In some embodiments, the Fc region comprises modification to reduce / prevent an Fc-mediated function.

[0707] In some embodiments, the Fc region comprises modification to reduce / prevent ADCC. In some embodiments, the Fc region comprises modification to reduce / prevent ADCP. In some embodiments, the Fc region comprises modification to reduce / prevent CDC. An antigen-binding molecule comprising an Fc region comprising modification to reduce / prevent an Fc-mediated function (e.g. ADCC, ADCP, CDC) induces an reduced level of the relevant effector function as compared to an antigen-binding molecule comprising the corresponding unmodified Fc region.

[0708] In some embodiments, the Fc region comprises modification to reduce / prevent binding to an Fc receptor. In some embodiments, the Fc region comprises modification to reduce / prevent binding to an Fey receptor. In some embodiments, the Fc region comprises modification to reduce / prevent binding to one or more of FcyRI, FcyRlla, FcyRllb, FcyRllc, FcyRllla and FcyRlllb. In some embodiments, the Fc region comprises modification to reduce / prevent binding to FcyRllla. In some embodiments, the Fc region comprises modification to reduce / prevent binding to FcyRlla. In some embodiments, the Fc region comprises modification to reduce / prevent binding to FcyRllb. In some embodiments, the Fc region comprises modification to reduce / prevent binding to a complement protein. In some embodiments, the Fc region comprises modification to reduce / prevent binding to C1q. In some embodiments, the Fc region comprises modification to reduce / prevent glycosylation of the amino acid residue corresponding to N297.

[0709] In some embodiments, the Fc region is not able to induce one or more Fc-mediated functions ( / .e. lacks the ability to elicit the relevant Fc-mediated function(s)). Accordingly, antigen-binding molecules comprising such Fc regions also lack the ability to induce the relevant function(s). Such antigen-binding molecules may be described as being devoid of the relevant function(s).

[0710] In some embodiments, the Fc region is not able to induce ADCC. In some embodiments, the Fc region is not able to induce ADCP. In some embodiments, the Fc region is not able to induce CDC. In some embodiments, the Fc region is not able to induce ADCC and / or is not able to induce ADCP and / or is not able to induce CDC.

[0711] In some embodiments, the Fc region is not able to bind to an Fc receptor. In some embodiments, the Fc region is not able to bind to an Fey receptor. In some embodiments, the Fc region is not able to bind to one or more of FcyRI, FcyRlla, FcyRllb, FcyRllc, FcyRllla and FcyRlllb. In some embodiments, the Fc region is not able to bind to FcyRllla. In some embodiments, the Fc region is not able to bind to FcyRlla. In some embodiments, the Fc region is not able to bind to FcyRllb. In some embodiments, the Fc region is not able to bind to FcRn. In some embodiments, the Fc region is not able to bind to a complement protein. In some embodiments, the Fc region is not able to bind to C1q. In some embodiments, the Fc region is not glycosylated at the amino acid residue corresponding to N297. In some embodiments, the Fc region comprises modification corresponding to N297A or N297Q or N297G as described in Leabman et al., MAbs. (2013) 5:896-903. In some embodiments, the Fc region comprises modification corresponding to L235E as described in Alegre et al., J Immunol. (1992) 148:3461-3468. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions L234A / L235A or F234A / L235A as described in Xu et al., Cell Immunol. (2000) 200:16-26. In some embodiments, the Fc region comprises modification corresponding to P329A or P329G as described in Schlothauer et al., Protein Engineering, Design and Selection (2016), 29(10):457-466. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions L234A / L235A / P329G as described in Lo et al. J. Biol. Chem (2017) 292(9):3900-3908. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions described in Rother et al., Nat Biotechnol. (2007) 25:1256-1264. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions S228P / L235E as described in Newman et al., Clin. Immunol. (2001 ) 98:164-174. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions H268QA / 309L / A330S / P331S as described in An et al., MAbs. (2009) 1 :572-579. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions

[0712] V234A / G237A / P238S / H268A / V309L / A330S / P331S as described in Vafa et al., Methods. (2014) 65:114- 126. In some embodiments, the Fc region comprises modification corresponding to the combination of substitutions L234A / L235E / G237A / A330S / P331S as described in US 2015 / 0044231 A1.

[0713] The combination of substitutions ‘L234A / L235A and corresponding substitutions (such as e.g. F234A / L235A in human lgG4) are known to disrupt binding of Fc to Fey receptors and inhibit ADCC, ADCP, and also to reduce C1q binding and thus CDC (Schlothauer et al., Protein Engineering, Design and Selection (2016), 29(10):457^466, hereby incorporated by reference in entirety). The substitutions ‘P329G’ and ‘P329A’ reduce C1q binding (and thereby CDC). Substitution of ‘N297’ with ‘A’, ‘G’ or ‘Q’ is known to eliminate glycosylation, and thereby reduce Fc binding to C1q and Fey receptors, and thus CDC and ADCC. Lo et al. J. Biol. Chem (2017) 292(9):3900-3908 (hereby incorporated by reference in its entirety) reports that the combination of substitutions L234A / L235A / P329G eliminated complement binding and fixation as well as Fc y receptor dependent, antibody-dependent, cell-mediated cytotoxicity in both murine lgG2a and human lgG1.

[0714] The combination of substitutions L234A / L235E / G237A / A330S / P331S in lgG1 Fc is disclosed in US 2015 / 0044231 A1 to abolish induction of phagocytosis, ADCC and CDC.

[0715] In some embodiments, the Fc region comprises modification corresponding to the substitution S228P as described in Silva et al., J Biol Chem. (2015) 290(9):5462-5469. The substitution S228P in lgG4 Fc reduces Fab-arm exchange (Fab arm exchange can be undesirable).

[0716] In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235A. In some embodiments, the Fc region comprises modification corresponding to corresponding to the substitution P329G. In some embodiments, the Fc region comprises modification corresponding to corresponding to the substitution N297Q. In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235A / P329G.

[0717] In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235A / P329G / N297Q.

[0718] In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235E / G237A / A330S / P331S.

[0719] In some embodiments, the Fc region comprises modification corresponding to corresponding to the substitution S228P, e.g. in lgG4.

[0720] In some embodiments - particularly embodiments in which the antigen-binding molecule (e.g. an antigenbinding moiety thereof) is a multispecific (e.g. bispecific) antigen-binding molecule - the antigen-binding molecule comprises an Fc region comprising modification in one or more of the CH2 and CH3 regions promoting association of the Fc region. Recombinant co-expression of constituent polypeptides of an antigen-binding molecule and subsequent association leads to several possible combinations. To improve the yield of the desired combinations of polypeptides in antigen-binding molecules in recombinant production, it is advantageous to introduce in the Fc regions modification(s) promoting association of the desired combination of heavy chain polypeptides. Modifications may promote e.g. hydrophobic and / or electrostatic interaction between CH2 and / or CH3 regions of different polypeptide chains. Suitable modifications are described e.g. in Ha et al., Front. Immnol (2016) 7:394, which is hereby incorporated by reference in its entirety.

[0721] In some embodiments, the antigen antigen-binding molecule of the present disclosure (e.g. an antigenbinding moiety thereof) comprises an Fc region comprising paired substitutions in the CH3 regions of the Fc region according to one of the following formats, as shown in Table 1 of Ha et al., Front. Immnol (2016) 7:394: KiH, KiHs.s, HA-TF, ZW1 , 7.8.60, DD-KK, EW-RVT, EW-RVTS.S, SEED or A107.

[0722] Polypeptides and particular exemplary antigen-binding moieties

[0723] The present disclosure also provides polypeptide constituents of antigen-binding moieties. The polypeptides may be provided in isolated or substantially purified form.

[0724] The antigen-binding moieties of the antigen-binding molecule of the present disclosure may be, or may comprise, a complex of polypeptides.

[0725] In the present specification where a polypeptide comprises more than one domain or region, it will be appreciated that the plural domains / regions are preferably present in the same polypeptide chain. That is, the polypeptide comprising more than one domain or region is a fusion polypeptide comprising the domains / regions. In some embodiments a polypeptide according to the present disclosure comprises, or consists of, a VH as described herein. In some embodiments a polypeptide according to the present disclosure comprises, or consists of, a VL as described herein.

[0726] In some embodiments, the polypeptide additionally comprises one or more antibody heavy chain constant regions (CH). In some embodiments, the polypeptide additionally comprises one or more antibody light chain constant regions (CL). In some embodiments, the polypeptide comprises a CH1, CH2 region and / or a CH3 region of an immunoglobulin (Ig).

[0727] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the polypeptide comprises a CH1 region as described herein. In some embodiments, the polypeptide comprises a CH1-CH2 hinge region as described herein. In some embodiments, the polypeptide comprises a CH2 region as described herein. In some embodiments, the polypeptide comprises a CH3 region as described herein.

[0728] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin light chain constant sequence. In some embodiments, the polypeptide comprises a CL region as described herein.

[0729] In some embodiments, the polypeptide according to the present disclosure comprises a structure from N- to C-terminus according to one of the following:

[0730] (i) VH

[0731] (ii) VL

[0732] (iii) VH-CH1

[0733] (iv) VL-CL

[0734] (v) VL-CH1

[0735] (vi) VH-CL

[0736] (vii) VH-CH1-CH2-CH3

[0737] (viii) VL-CL-CH2-CH3

[0738] (ix) VL-CH1-CH2-CH3

[0739] (x) VH-CL-CH2-CH3

[0740] Also provided by the present disclosure are antigen-binding moieties composed of the polypeptides of the present disclosure. In some embodiments, the antigen-binding moiety of the present disclosure comprises one of the following combinations of polypeptides:

[0741] (A) VH + VL

[0742] (B) VH-CH1 + VL-CL

[0743] (C) VL-CH1 + VH-CL

[0744] (D) VH-CH1-CH2-CH3 + VL-CL

[0745] (E) VH-CL-CH2-CH3 + VL-CH1

[0746] (F) VL-CH1-CH2-CH3 + VH-CL

[0747] (G) VL-CL-CH2-CH3 + VH-CH1

[0748] (H) VH-CH1-CH2-CH3 + VL-CL-CH2-CH3 (I) VH-CL-CH2-CH3 + VL-CH1-CH2-CH3

[0749] In some embodiments, the antigen-binding moiety comprises more than one polypeptide of the combinations shown in (A) to (I) above. By way of example, with reference to (D) above, in some embodiments, the antigen-binding moiety comprises two polypeptides comprising the structure VH-CH1- CH2-CH3, and two polypeptides comprising the structure VL-CL.

[0750] In some embodiments, the antigen-binding moiety of the present disclosure comprises one of the following combinations of polypeptides:

[0751] (J) VH (anti-CNX) + VL (anti-CNX)

[0752] (K) VH (anti-CNX)-CHI + VL (anti-CNX)-CL

[0753] (L) VL (anti-CNX)-CHI + VH (anti-CNX)-CL

[0754] (M) VH (anti-CNX)-CH1-CH2-CH3 + VL (anti-CNX)-CL

[0755] (N) VH (anti-CNX)-CL-CH2-CH3 + VL (anti-CNX)-CHI

[0756] (O) VL (anti-CNX)-CH1-CH2-CH3 + VH (anti-CNX)-CL

[0757] (P) VL (anti-CNX)-CL-CH2-CH3 + VH (anti-CNX)-CHI

[0758] (Q) VH (anti-CNX)-CH1-CH2-CH3 + VL (anti-CNX)-CL-CH2-CH3

[0759] Wherein: ‘VH(anti-CNX)’ refers to the VH of an antigen-binding moiety capable of binding to CNX as described herein, e.g. as defined in one of (1 ) to (75); and ‘VL(anti-CNX)’ refers to the VL of an antigenbinding moiety capable of binding to CNX as described herein, e.g. as defined in one of (76) to (164).

[0760] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:1 , 17, 32, 47, 60, 82, 85, 94, 107, 121, 131 , 145, 155, 165, 184, 198, 210, 221 , or 242.

[0761] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:9, 24, 40, 52, 67, 72, 77, 88, 100, 114, 124, 138, 152, 157, 170, 178, 191 , 204, 215, 228, 234 or 247.

[0762] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:273, 276, 279, 282, 285, 290, 292, 295, 298, 301, 304, 307, 310, 313, 317, 320, 323, 326, 330, 274, 277, 280, 283, 286, 291 , 293, 296, 299, 302, 305, 308, 311, 314, 318, 321 , 324, 327, or 331.

[0763] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:275, 278, 281 , 284, 287, 288, 289, 294, 297, 300, 303, 306, 309, 312, 315, 316, 319, 322, 325, 328, 329 or 332.

[0764] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide or polypeptides comprising a VH region comprising the heavy chain CDRs, and a VL region comprising the light chain CDRs, of a clone selected from 1 D3, 1 D6, 1 E1 , 1 E6, 2C6, 2H6, 3D1 , 2G9, 2G12, 2H5, 3F8, 3F9, 4G9, 5A3, 5E8, C001, C008, C010, C023, C025, C040, C046 and C117, as shown in Table A herein. That is, in some embodiments, the antigen-binding moiety comprises a polypeptide or polypeptides comprising: (i) a VH region comprising HC-CDR1 , HC-CDR2 and HC-CDR3 as indicated in column A of Table A, and (ii) a VL region comprising LC-CDR1 , LC-CDR2 and LC-CDR3 as indicated in column B of Table A, wherein the sequences of columns A and B are selected from the same row of Table A.

[0765] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide or polypeptides comprising a VH region comprising the heavy chain FRs, and a VL region comprising the light chain FRs, of a clone selected from 1 D3, 1 D6, 1 E1 , 1 E6, 2C6, 2H6, 3D1, 2G9, 2G12, 2H5, 3F8, 3F9, 4G9, 5A3, 5E8, C001, C008, C010, C023, C025, C040, C046 and C117, as shown in Table B herein. That is, in some embodiments, the antigen-binding moiety comprises a polypeptide or polypeptides comprising: (i) a VH region comprising HC-FR1, HC-FR2, HC-FR3 and HC-FR4 as indicated in column A of Table B, and (ii) a VL region comprising LC-FR1, LC-FR2, LC-FR3, and LC-FR4 as indicated in column B of Table B, wherein the sequences of columns A and B are selected from the same row of Table B.

[0766] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide or polypeptides comprising: (i) an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to an amino acid sequence indicated in column A of Table C, and (ii) an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to an amino acid sequence indicated in column B of Table C, wherein the sequences of columns A and B are selected from the same row of Table C.

[0767] In some embodiments, the antigen-binding moiety of the present disclosure comprises a polypeptide or polypeptides comprising a VH region and a VL region of a clone selected from 1D3, 1 D6, 1 E1, 1 E6, 2C6, 2H6, 3D1 , 2G9, 2G12, 2H5, 3F8, 3F9, 4G9, 5A3, 5E8, C001, C008, C010, C023, C025, C040, C046 and C117, as shown in Table C herein. That is, in some embodiments, the antigen-binding moiety comprises a polypeptide or polypeptides comprising: (i) an amino acid sequence indicated in column A of Table C, and (ii) an amino acid sequence indicated in column B of Table C, wherein the sequences of columns A and B are selected from the same row of Table C.

[0768] In some embodiments, the antigen-binding moiety of the present disclosure comprises: (i) a polypeptide comprising or consisting of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to an amino acid sequence indicated in column A of Table D, and (ii) a polypeptide comprising or consisting of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to an amino acid sequence indicated in column B of Table D, wherein the sequences of columns A and B are selected from the same row of Table D.

[0769] In some embodiments, the antigen-binding moiety of the present disclosure comprises the polypeptides of an antigen-binding moiety according to any one of [1] to

[0046] as detailed in Table D herein. That is, in some embodiments, the antigen-binding moiety comprises: (i) a polypeptide comprising or consisting of an amino acid sequence indicated in column A of Table D, and (ii) a polypeptide comprising or consisting of an amino acid sequence indicated in column B of Table D, wherein the sequences of columns A and B are selected from the same row of Table D.

[0770] Linker-payload moieties

[0771] Aspects and embodiments of the present disclosure relate to antigen-binding molecules comprising a linker-payload moiety. As used herein, a linker-payload moiety refers to a moiety comprising one or more payload moieties, and a linker moiety for linking the payload moiety(Zies) to the antigen-binding moiety of the antigen-binding molecule.

[0772] In some embodiments, a payload moiety according to the present disclosure comprises or consists of a cytotoxic agent. Payload moieties are described e.g. in Parslow et al., Biomedicines. 2016 Sep; 4(3):14, Goundry and Parker, Org. Process Res. Dev. (2022) 26, 8, 2121-2123, Fu et al., Signal Transduction and Targeted Therapy (2022) 7:93, Wang et al., Acta Pharmaceutica Sinica B (2023) 13 (10): 4025-4059 and Conilh et al., J. Hematol. & Oncol. (2023) 16:3, all of which are hereby incorporated by reference in their entirety.

[0773] In some embodiments, a payload moiety according to the present disclosure may comprise or consist of a microtubule-targeting agent, a DNA-targeting agent, an RNA-targeting agent, an immune systemactivating agent, an apoptosis-promoting agent, a metabolism-inhibiting agent, a proteasome inhibiting agent, and / or a protein-degrader agent.

[0774] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a microtubule-targeting agent. Microtubules play important roles in maintaining proper cellular morphology, signal transduction, organelle transportation, cell motility and cell division. Microtubules are formed of tubulin, and agents that disrupt the tubulin polymerization dynamics, resulting in cell cycle arrest and apoptosis. Tubulin inhibitors have a stronger toxicity to rapidly-dividing cancerous cells than slower- growing, non-cancerous cells. Microtubule-targeting agents include tubulin polymerization enhancers (e.g. auristatins, taxanes), and tubulin polymerization inhibitors (e.g. maytansinoids, colchicine).

[0775] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a maytansinoid, e.g. maytansine or a derivative thereof, e.g. mytansine, DM1 (mertansine) or DM4 (ravtansine). In some embodiments, the maytansinoid is not DM1. In some embodiments, the maytansinoid is not DM4. In some embodiments, a payload moiety comprises, or consists of colchicine or a derivative thereof.

[0776] In some embodiments, a payload moiety comprises, or consists of, an auristatin, e.g. a dolastatin 10 derivative, e.g. monomethyl auristatin E (MMAE), auristatin F, monomethyl auristatin F (MMAF), auristatin PE, auristatin PYE, PF-06380101 , auristatin F-hydroxypropylamide (AF-HPA) or azastatin. In some embodiments, the auristatin is not MMAE. In some embodiments, the auristatin is not MMAF. In some embodiments, a payload moiety comprises, or consists of, a halichondrin B derivative, e.g. eribulin. In some embodiments, a payload moiety comprises, or consists of, a tubulysin or a derivative thereof, e.g. tubulysin A, D, H, U or V. In some embodiments, a payload moiety comprises, or consists of, a cryptophycin or a derivative thereof, e.g. cryptophycin-1 , cryptophycin-52, cryptophycin-55 or cryptophycin-55gly. In some embodiments, a payload moiety comprises, or consists of, an EG5 ( / .e. kinesin / KSP / KIF11) inhibitor, e.g. ispinesib (SB715992) or a derivative thereof, or filanesib (ARRY-520) or a derivative thereof. In some embodiments, a payload moiety comprises, or consists of, a taxane, e.g. paclitaxel, docetaxel or cabazitaxel. In some embodiments, a payload moiety comprises, or consists of vinca alkaloid, e.g. vinblastine, vincristine, vindesine, vinorelbine or vinflunine. In some embodiments, a payload moiety comprises, or consists of hemiasterlin or a derivative thereof, e.g. hemiasterlin, hemisterlin A or HTI-286.

[0777] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a DNA-targeting agent. DNA-targeting agents include agents that directly or indirectly destroy DNA through introducing / promoting the formation of single- and / or double-strand breaks (e.g. enediynes, topoisomerase inhibitors), DNA alkylating agents (e.g. pyrrolo[2,1-c][1 ,4] benzodiazepines (PBD), indolinobenzodiazpines, duocarmycins), and DNA crosslinking agents (e.g. mitomycin C). In some embodiments, the DNA alkylating agent is not a duocarmycin. In some embodiments, the DNA alkylating agent is not D6.5. In some embodiments, the DNA alkylating agent is not a PBD.

[0778] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, an enediyne, e.g. a Cal-like enediyne or an anthraquinone fusion enediyne, e.g. calicheamicin yh, calicheamicin Oh or uncialamycin. In some embodiments, the enediyne is not calicheamicin. In some embodiments, a payload moiety comprises, or consists of, a topoisomerase inhibitor, e.g. a TOP1 or TOP2 inhibitor, e.g. camptothecin or a derivative thereof, e.g. SN-38, exatecan, exatecan mesylate (DX- 8951f), A / -glycyl-exatecan or deruxtecan (Dxd); e.g. an anthracycline, e.g. doxorubicin, daunorubicin, epirubicin, PNU-159682 or idarubicin. In some embodiments the topoisomerase inhibitor is not SN-38. In some embodiments the topoisomerase inhibitor is not doxorubicin.

[0779] In some embodiments, a payload moiety comprises, or consists of, a pyrrolo[2,1-c][1,4] benzodiazepine (PBD) dimer, or a derivative thereof, e.g. a PDB, KMR-28-39, SJG-136 SGD-1882 or SG3199 dimer. In some embodiments, a payload moiety comprises, or consists of, indolinobenzodiazpine (IGN; monoimine) or a derivative thereof. In some embodiments, a payload moiety comprises, or consists of, a pyridinobenzodiazepine (PDD) dimer, or a derivative thereof, e.g. a PDD or FGX5-67 dimer. In some embodiments, a payload moiety comprises, or consists of, a duocarmycin or a derivative thereof, e.g. duocarmycin A, CC1065, duocarmycin SA, DUBA, seco-DIBA or seco-CBI.

[0780] In some embodiments, a payload moiety comprises, or consists of, mitomycin C.

[0781] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a RNA-targeting agent. Small molecule inhibitors that target RNA can kill both dividing and dormant tumor cells. RNA-targeting agents include RNA splicing inhibitors (e.g. thailanstatin and derivatives thereof) and RNA polymerase II inhibitors (e.g. amatoxins, RNA polymerase ll-IN-2). In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, thailanstatin or a derivative thereof, e.g. thailanstatin A, thailanstatin B, thailanstatin C or FR901464. In some embodiments, a payload moiety comprises, or consists of, an amatoxin, e.g. a-amanitin or p-amanitin. In some embodiments, a payload moiety comprises, or consists of, RNA polymerase ll-IN-2.

[0782] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, an immune system-activating agent. Immune-stimulating antibody conjugates (ISACs) employ small molecule-based engagement of the innate and / or adaptive immune systems. A variety of immune- modulating payloads are in development, including Toll-like receptor (TLR) agonists, stimulator of interferon genes (STING) agonists and glucocorticoid receptor modulators (GRMs). In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a TLR agonist, e.g. an agonist of TLR7, TLR8 or TLR9. In some embodiments, a payload moiety comprises, or consists of, a STING agonist, e.g. a cyclic dinucleotide (CDN; e.g. 2,3 cGAMP; 3,3 cGAMP; c-di-GMP or c-di-AMP) or a benzimidazole. In some embodiments, a payload moiety comprises, or consists of, a glucocorticoid receptor modulator, e.g. dexamethasone or a derivative thereof.

[0783] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, an apoptosis-promoting agent. Anti-apoptotic proteins such as Bcl-xL can play important roles in tumorigenesis, metastasis and drug resistance. In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a Bcl-xL inhibitor. In some embodiments, a payload moiety comprises, or consists of, ABT-737.

[0784] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a metabolism-inhibiting agent. Metabolism-inhibiting agents such as niacinamide phosphate ribose transferase (NAMPT) inhibitors control the concentration of NAD+ within cells, inducing energy crisis and cell death, and antifolate antimetabolites such as methotrexate, that inhibit dihydrofolate reductase (DHFR) and thereby DNA synthesis. In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a NAMPT inhibitor, e.g. FK-866 or A-1293201. In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a DHFR inhibitor, e.g. methotrexate of a derivative thereof.

[0785] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a proteasome-inhibiting agent. Proteasome-inhibiting agents include carmaphycins. In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a carmaphycin or a derivative thereof, e.g. carmaphycin A or carmaphycin B.

[0786] In some embodiments, a payload moiety according to the present disclosure comprises, or consists of, a protein-degrader agent, e.g. a targeted protein degrader. In some embodiments, a payload moiety according to the present disclosure comprises or consists of a proteolysis targeting chimera (PROTAC). PROTACs may comprise two ligands joined by a linker; one ligand recruits the protein of interest ( / .e. the protein to be degraded) and the other ligand recruits the E3 ubiquitin ligase. Once the PROTAC molecule binds to the target protein and the E3 ubiquitin ligase, the E3 ligase may ubiquitinate the target protein and initiate the degradation process.

[0787] In some embodiments, the payload moiety is monomethylauristatin E (MMAE). In some embodiments, the payload moiety is deruxtecan (Dxd). In some embodiments, the payload moiety is Auristatin F.

[0788] In some embodiments, the payload moiety is not calicheamicin, DM1 , DM4, monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), SN-38, doxorubicin, duocarmycin, D6.5 or PBD.

[0789] A linker moiety according to the present disclosure may be any moiety suitable for linking the payload moiety to the antigen-binding moiety of the antigen-binding molecule of the present disclosure.

[0790] Accordingly, they generally comprise a group enabling connection to the payload moiety, a group enabling conjugation to the antigen-binding moiety of the antigen-binding molecule, and a linker core.

[0791] Linker moieties are described e.g. in Su et al., Acta Pharmaceutica Sinica B (2021 ) 11(12): 3889-3907, Fu et al., Signal Transduction and Targeted Therapy (2022) 7:93,

[0792] A linker moiety according to the present disclosure may be a cleavable linker moiety or a non-cleavable moiety.

[0793] Cleavable linkers typically utilise differences between the environment of systemic circulation and that in cancer cells / the tumor microenvironment to release the payload moiety in a targeted manner. Cleavable linkers include chemical cleavage linkers (e.g. acid-cleavable linkers, GSH-cleavable linkers, Fe(ll)- cleavable linkers) and enzyme cleavage linkers (e.g. cathepsin-cleavable linkers, glycosidase-cleavable linkers, phosphatase-cleavable linkers, sulfatase-cleavable linkers).

[0794] In some embodiments, a linker moiety according to the present disclosure is a chemical cleavage linker. In some embodiments, a linker moiety according to the present disclosure is an enzyme cleavage linker. In some embodiments, a linker moiety is an acid-cleavable linker, e.g. comprising a hydrazone group (e.g. a 6-maleimidocaproylhydrazone linker or a (4-(4-acetylphenoxy)butanoic acid) hydrazaone linker), a carbonate group or a silyl ether group. In some embodiments, a linker moiety is a GSH-cleavable linker, e.g. comprising a disulfide group. In some embodiments, a linker moiety is a Fe(ll)-cleavable linker, e.g. comprising a 1,2,4-trioxolane group. In some embodiments, a linker moiety is a cathepsin-cleavable linker, e.g. comprising a dipeptide (e.g. a valine-citrulline linker, a phenylalanine-lysine linker or a valine- alanine linker), a triglycyl peptide (CX), a tetrapeptide (e.g. a glycine-glycine-phenylalanine-glycine (GGFG) linker) or a cBu-Cit group. In some embodiments, a linker moiety is a glucuronidase-cleavable linker, e.g. comprising a p-glucuronide group. In some embodiments, a linker moiety is a glycosidase- cleavable linker, e.g. comprising a p-galactoside group. In some embodiments, a linker moiety is a phosphatase-cleavable linker, e.g. comprising a pyrophosphate group. In some embodiments, a linker moiety is a sulfatase-cleavable linker, e.g. comprising an arylsulfate group. In some embodiments, a linker moiety is a photo-responsive linker, e.g. comprising a heptamethine cyanine fluorophore group, an O-nitrobenzyl group or a PC4AP group. In some embodiments, a linker moiety is a biorthogonal cleavable linker, e.g. comprising a dsProc group. In some embodiments, a linker moiety comprises a benzyl group. In some embodiments, a linker moiety comprises a bis-functional electrophilic group, e.g. a bis-sulfone group or a bis-sulfone bis-alkylating group.

[0795] Non-cleavable linkers remain inert in common chemical and enzymatic environments in the body, with the payload moiety being released following processing of the ADC by cellular lysosomal proteases. Non- cleavable linkers include linkers comprising thioether or maleimidocaproyl (MC) groups.

[0796] In some embodiments, a linker moiety is a thioether linker. In some embodiments, a linker moiety is a maleimidocaproyl linker, e.g. comprising a 2-(maleimidomethyl)-1 ,3-dioxane (MD) group or a Mal-PAB group. In some embodiments, a linker moiety comprises a polyethylene glycol (PEG) group and an alkyne, triazole or piperazine group.

[0797] In some embodiments, a linker-payload moiety has a maleimide group for linkage to the antigen-binding moiety. In some embodiments, a linker-payload moiety has a thiol group for linkage to the antigen-binding moiety. In some embodiments, a linker-payload moiety according to the present disclosure has an amino (-NH2) group for linkage to the antigen-binding moiety, for example by enzymatic conjugation. In some of these embodiments, enzymatic conjugation with microbial transglutaminase may be used to conjugate the linker-payload moiety to the antigen-binding moiety.

[0798] In some embodiments, a linker moiety further comprises a spacer moiety. Spacer moieties are sometimes required due to the bulky nature of payload moieties. Commonly employed spacer moieties include paraaminobenzyl carbamate (PABC), hemiaminal groups, PEG groups, polar acyl sulfamide groups, polar carbamoyl sulfamide groups and HydraSpace (described e.g. in Verkade et al., Antibodies (Basel) (2018) 7(1 ):12 and WO 2016 / 053107 A1 , both of which are hereby incorporated by reference in their entirety). Para-aminobenzyl (PAB) or derivatives thereof (e.g. PABC) are commonly employed as a spacer moiety in cathepsin-cleavable dipeptide linkers, p-glucuronidase-cleavable linkers, p-galactosidase-cleavable linkers and phosphatase cleavable linkers. In some embodiments, a linker moiety comprises a para- aminobenzyl (PAB) as a spacer moiety.

[0799] In some embodiments, the linker moiety is MC-Val-Cit-PAB. In some embodiments, the linker moiety is MC-GGFG. In some embodiments, the linker moiety is benzyl-guanine. In some embodiments, a linker-payload moiety according to the present disclosure comprises, or consists of, ozogamicin, vedotin, emtansine, deruxtecan, govitecan, mafodotin, tesirine or soravtansine. The structures of the linker-payload moieties referred to in the preceding sentence are shown e.g. in Lim et al., Drugs Drug Candidates (2023) 2(2), 377-421, which is hereby incorporated by reference in its entirety (see in particular Figure 4).

[0800] In some embodiments, a linker-payload moiety comprises MC-Val-Cit-PAB-MMAE. In some embodiments, a linker-payload moiety comprises MC-GGFG-Dxd. In some embodiments, a linker payload moiety comprises benzyl-guanine-Auristatin F.

[0801] In some embodiments, a linker-payload moiety according to the present disclosure comprises more than one payload. This allows for a higher drug to antibody ratio (DAR) than when the linker-payload moiety comprises a single payload. Where the linker-payload moieties are attached to specific / engineered sites on the antigen-binding moieties, such linker-payload moieties allow for higher specific drug antibody ratios. For example, where two conjugation sites are available, a DAR of 2, 4, 6 or 8 can be achieved.

[0802] The antigen-binding molecules of the present disclosure may have a given drug to antibody ratio (DAR). As used herein, “drug to antibody ratio” or “drug antibody ratio” (DAR) refers to the average number of payload moieties conjugated to the antigen-binding moiety (e.g. a CNX-binding moiety). In some embodiments, the antigen-binding molecules have a DAR of between 0 and 20, e.g. between 0 and 15, between 1 and 15, between 0 and 10, between 1 and 10, between 0 and 8, between 1 and 8, between 2 and 8, between 4 and 8, between 6 and 8, between 0 and 6, between 1 and 6, between 2 and 6, between 4 and 6, between 0 and 4, between 1 and 4, between 2 and 4. In some embodiments, the antigen-binding molecules have a DAR of between 1 and 8. In some embodiments, the antigen-binding molecules have a DAR of e.g. 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10. In some embodiments, the antigen-binding molecules have a DAR of 1. In some embodiments, the antigen-binding molecules have a DAR of 4. In some embodiments, the antigen-binding molecules have a DAR of 8.

[0803] By way of illustration, in the antibody-drug conjugates of Example 2: 1 E1-MC-Val-Cit-PAB-MMAE has a DAR of 4, i.e. there is an average of four MMAE molecules conjugated to a single 1 E1 antibody clone; 1E1-MC-GGFG-Dxd has a DAR of 8, i.e. there is an average of eight Dxd molecules conjugated to a single 1 E1 antibody clone; scFV(C008)-SNAP-Auristatin F has a DAR or 1 , i.e. there is an average of one Auristatin F molecule conjugated to each scFv(C008) antigen-binding moiety.

[0804] Additional sequences

[0805] The antigen-binding molecules of the present disclosure and their constituent polypeptides may additionally comprise further amino acids or sequences of amino acids.

[0806] The polypeptides of the present disclosure may comprise one or more linker amino acid sequences between amino acid sequences. A linker sequence may be provided at one or both ends of one or more of a VH, VL, CH1-CH2 hinge region, CH2 region and a CH3 region of the antigen-binding molecule / polypeptide. Amino acid linker sequences are known to the skilled person, and are described, for example in Chen et al., Adv Drug Deliv Rev (2013) 65(10): 1357-1369, which is hereby incorporated by reference in its entirety. In some embodiments, a linker sequence may be a flexible linker sequence. Flexible linker sequences allow for relative movement of the amino acid sequences which are linked by the linker sequence. Flexible linkers are known to the skilled person, and several are identified in Chen et al., Adv Drug Deliv Rev (2013) 65(10): 1357-1369. Flexible linker sequences often comprise high proportions of glycine and / or serine residues.

[0807] In some embodiments, the linker sequence comprises at least one glycine residue and / or at least one serine residue. In some embodiments, the linker sequence comprises or consists of glycine and serine residues. In some embodiments, the linker sequence has the structure: (GxS)n (SEQ ID NO;410 and 411) or (GxS)nGm (SEQ ID NO:412 and 413); wherein G = glycine, S = serine, x = 3 or 4, n = 2, 3, 4, 5 or 6, and m = 0, 1 , 2 or 3. In some embodiments, the linker sequence comprises one or more (e.g. 1 , 2, 3, 4, 5 or 6) copies (e.g. in tandem) of the sequence motif G4S (SEQ ID NO:414). In some embodiments, the linker sequence comprises or consists of (G4S)4 (SEQ ID NO:415) or (G4S)e (SEQ ID NO:416). In some embodiments, the linker sequence has a length of 1-2, 1-3, 1-4, 1-5, 1-10, 1-15, 1-20, 1-25, or 1-30 amino acids.

[0808] The antigen-binding molecules of the present disclosure and their constituent polypeptides may additionally comprise amino acid sequence(s) to facilitate expression, folding, trafficking, processing, purification, or detection of the antigen-binding molecule / polypeptide. For example, antigen-binding molecules and polypeptides of the present disclosure may additionally comprise a sequence of amino acids forming a detectable moiety, e.g. as described hereinbelow.

[0809] The antigen-binding molecules and polypeptides of the present disclosure and their constituent polypeptides may additionally comprise a signal peptide (also known as a leader sequence or signal sequence). Signal peptides normally consist of a sequence of 5-30 hydrophobic amino acids, which form a single alpha helix. Secreted proteins and proteins expressed at the cell surface often comprise signal peptides.

[0810] The signal peptide may be present at the N-terminus of the polypeptide, and may be present in the newly synthesised polypeptide. The signal peptide provides for efficient trafficking and secretion of the polypeptide. Signal peptides are often removed by cleavage, and thus are not comprised in the mature polypeptide.

[0811] Signal peptides are known for many proteins, and are recorded in databases such as GenBank, UniProt, Swiss-Prot, TrEMBL, Protein Information Resource, Protein Data Bank, Ensembl, and InterPro, and / or can be identified / predicted e.g. using amino acid sequence analysis tools such as SignalP (Petersen et al., 2011 Nature Methods 8: 785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24: 2172- 2176). The antigen-binding moieties of the present disclosure and their constituent polypeptide(s) may comprise amino acid sequence(s) to facilitate conjugation to a payload moiety (e.g. a linker-payload moiety). For example, the antigen-binding moiety of the present disclosure or a constituent polypeptide(s) may comprise a “self-labelling protein”. As used herein, a self-labelling protein refers to a polypeptide which upon incubation with a specific partner ligand, reacts with the specific partner ligand to form a covalent attachment. Self-labelling proteins may be fused to an antigen-binding moiety or a constituent polypeptide thereof, e.g. expressed as a fusion protein. A linker-payload moiety may comprise a specific partner ligand to a given self-labelling protein, e.g. O6-benzylguanine, benzylcytosine, chloroalkane, or derivatives thereof. Self-labeling proteins are known in the art, for example, SNAP-tag, CLIP-tag and HaloTag. Selflabelling proteimligand pairings include e.g. SNAP-tag and O6-benzylguanine or derivatives thereof, CLIP- tag and benzylcytosine or derivatives thereof, HaloTag and chloroalkane or derivatives thereof. Selflabelling proteins are described in Wilhelm et al., 2021 Biochemistry 60(33): 2560-2575. In some embodiments, the antigen-binding moiety further comprises a SNAP protein. In some embodiments the SNAP protein comprises or consists of an amino acid sequence having at least 70%, preferably one of 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 419.

[0812] Labels and conjugates

[0813] In some embodiments, the antigen-binding molecules of the present disclosure additionally comprise a detectable moiety.

[0814] In some embodiments, the antigen-binding molecule comprises a detectable moiety, e.g. a fluorescent label, phosphorescent label, luminescent label, immuno-detectable label (e.g. an epitope tag), radiolabel, chemical, nucleic acid or enzymatic label. The antigen-binding molecule may be covalently or non- covalently labelled with the detectable moiety.

[0815] Fluorescent labels include e.g. fluorescein, rhodamine, allophycocyanin, eosine and NDB, green fluorescent protein (GFP), chelates of rare earths such as europium (Eu), terbium (Tb) and samarium (Sm), tetramethyl rhodamine, Texas Red, 4-methyl umbelliferone, 7-amino-4-methyl coumarin, Cy3, and Cy5. Radiolabels include radioisotopes such as Hydrogen3, Sulfur35, Carbon14, Phosphorus32, Iodine123, Iodine125, Iodine126, Iodine131, Iodine133, Bromine77, Technetium99"1, Indium111, Indium113"1, Gallium67, Gallium68, Ruthenium95, Ruthenium97, Ruthenium103, Ruthenium105, Mercury207, Mercury203, Rhenium99"1, Rhenium101, Rhenium105, Scandium47, Tellurium121"1, Tellurium122"1, Tellurium125"1, Thulium165, Thuliuml167, Thulium168, Copper67, Fluorine18, Yttrium90, Palladium100, Bismuth217and Antimony211. Luminescent labels include as radioluminescent, chemiluminescent (e.g. acridinium ester, luminol, isoluminol) and bioluminescent labels. Immuno-detectable labels include haptens, peptides / polypeptides, antibodies, receptors and ligands such as biotin, avidin, streptavidin or digoxigenin. Nucleic acid labels include aptamers.

[0816] In some embodiments, the antigen-binding molecule or a constituent polypeptide thereof comprises an epitope tag, e.g. a His, (e.g. 6XHis), FLAG, c-Myc, StrepTag, haemagglutinin, calmodulin-binding protein (CBP), glutathione-s-transferase (GST), maltose-binding protein (MBP), thioredoxin, S-peptide, T7 peptide, SH2 domain, avidin, streptavidin, and haptens (e.g. biotin, digoxigenin, dinitrophenol), optionally at the N- or C- terminus of the antigen-binding molecule / polypeptide.

[0817] In some embodiments, the antigen-binding molecule or a constituent polypeptide thereof comprises a moiety having a detectable activity, e.g. an enzymatic moiety. Enzymatic moieties include e.g. luciferases, glucose oxidases, galactosidases (e.g. beta-galactosidase), glucorinidases, phosphatases (e.g. alkaline phosphatase), peroxidases (e.g. horseradish peroxidase) and cholinesterases.

[0818] Functional properties of the antigen-binding moieties and antigen-binding molecules

[0819] The antigen-binding moieties and antigen-binding molecules described herein may be characterised by reference to certain functional properties.

[0820] In some embodiments, an antigen-binding molecule / moiety described herein may possess one or more of the following properties: binds to CNX (e.g. human CNX and / or mouse CNX); binds to CRT (e.g. human CRT); binds cross-reactively to CNX (e.g. human CNX and / or mouse CNX) and CRT (e.g. human CRT); reduces a function of CNX / CRT and / or a function of a complex comprising CNX / CRT ; reduces or inhibits extracellular matrix degradation (e.g. collagen and / or gelatin degradation); reduces or inhibits extracellular matrix degradation activity of a cell characterised by CNX expression; reduces or inhibits extracellular matrix degradation activity of a cancer cell; reduces or inhibits extracellular matrix degradation by a cell characterised by CNX expression; reduces or inhibits extracellular matrix degradation by a cancer cell; reduces oxireductase activity; reduces disulfide bond reductase activity; increases killing of cells expressing CNX / CRT; increases ADCC of cells expressing CNX / CRT; inhibits tumor growth; reduces or prevents metastasis of a cancer; increases survival of subjects having a cancer; and / or reduces the pathology of a disease / condition characterised by ECM degradation in a subject.

[0821] More specifically, in some embodiments, an antigen-binding molecule described herein may possess one of more of the following properties: binds to cells expressing CNX / CRT; is capable of being internalised by cells expressing CNX / CRT; increases killing of cells expressing CNX / CRT; increases killing of cancer cells (e.g. cancer cells expressing CNX / CRT); inhibits tumor growth and / or reduces tumor size / volume; reduces or prevents metastasis of a cancer; and / or increases survival of subjects having a cancer. It will be appreciated that a given antigen-binding molecule / moiety may display more than one of the properties recited in the preceding paragraph. A given antigen-binding molecule / moiety may be evaluated for the properties recited in the preceding paragraph using suitable assays. For example, the assays may be e.g. in vitro assays, optionally cell-based assays or cell-free assays. In some embodiments, the assays may be e.g. in vivo assays, i.e. performed in non-human animals. In some embodiments, the assays may be e.g. ex vivo assays, i.e. performed using cells / tissue / an organ obtained from a subject.

[0822] Where assays are cell-based assays, they may comprise treating cells with a given antigen-binding molecule in order to determine whether the antigen-binding molecule / moiety displays one or more of the recited properties. Assays may employ species labelled with detectable entities in order to facilitate their detection. Assays may comprise evaluating the recited properties following treatment of cells separately with a range of quantities / concentrations of a given antigen-binding molecule / moiety (e.g. a dilution series). It will be appreciated that the cells preferably express the target antigen for the antigen-binding molecule (i.e. CNX / CRT).

[0823] Analysis of the results of such assays may comprise determining the concentration at which 50% of the maximal level of the relevant activity is attained. The concentration of a given agent at which 50% of the maximal level of the relevant activity is attained may be referred to as the ‘half-maximal effective concentration’ of the agent in relation to the relevant activity, which may also be referred to as the ‘EC50’. By way of illustration, the EC50 of a given antigen-binding molecule for binding to human CNX may be the concentration of the antigen-binding molecule at which 50% of the maximal level of binding to human CNX is achieved.

[0824] Depending on the property, the EC50 may also be referred to as the ‘half-maximal inhibitory concentration’ or ‘IC50’, this being the concentration of the agent at which 50% of the maximal level of inhibition of a given property is observed.

[0825] The antigen-binding molecules described herein bind to CNX. In some embodiments, the antigen-binding molecules bind to CRT. The antigen-binding molecules and antigen-binding domains described herein preferably display specific binding to the relevant target antigen (e.g. CNX). As used herein, ‘specific binding’ refers to binding which is selective for the antigen, and which can be discriminated from nonspecific binding to non-target antigen. An antigen-binding molecule / domain that specifically binds to a target molecule preferably binds the target with greater affinity, and / or with greater duration than it binds to other, non-target molecules.

[0826] The ability of a given polypeptide to bind specifically to a given molecule can be determined by analysis according to methods known in the art, such as by ELISA, Surface Plasmon Resonance (SPR; see e.g. Hearty et al., Methods Mol Biol (2012) 907:411-442), Bio-Layer Interferometry (see e.g. Lad et al., (2015) J Biomol Screen 20(4): 498-507), flow cytometry, or by a radiolabeled antigen-binding assay (RIA) enzyme-linked immunosorbent assay. Through such analysis binding to a given molecule can be measured and quantified. In some embodiments, the binding may be the response detected in a given assay.

[0827] In some embodiments, the extent of binding of the antigen-binding molecule to a non-target molecule is less than about 10% of the binding of the antibody to the target molecule as measured, e.g. by ELISA, SPR, Bio-Layer Interferometry or by RIA. Alternatively, binding specificity may be reflected in terms of binding affinity where the antigen-binding molecule binds with a dissociation constant (KD) that is at least 0.1 order of magnitude ( / .e. 0.1 x 10n, where n is an integer representing the order of magnitude) greater than the KD of the antigen-binding molecule towards a non-target molecule. This may optionally be one of at least 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1 .0, 1 .5, or 2.0.

[0828] The affinity of binding to a given target antigen for an antigen-binding molecule described herein may be determined by Bio-Layer Interferometry, e.g. as described in the Examples of the present disclosure.

[0829] In some embodiments, the antigen-binding molecule described herein binds to CNX with an affinity in the micromolar range, i.e. KD = 9.9 X 10-4to 1 X 10-6M. In some embodiments, the antigen-binding molecule described herein binds to CNX with sub-micromolar affinity, i.e. KD < 1 x 10-6M. In some embodiments, the antigen-binding molecule described herein binds to CNX with an affinity in the nanomolar range, i.e. KD = 9.9 x 10-7to 1 x 10-9M. In some embodiments, the antigen-binding molecule described herein binds to CNX with sub-nanomolar affinity, i.e. KD < 1 x 10‘9M. In some embodiments, the antigen-binding molecule described herein binds to CNX with an affinity in the picomolar range, i.e. KD = 9.9 x 10-10to 1 x 10-12M. In some embodiments, the antigen-binding molecule described herein binds to CNX with sub- picomolar affinity, i.e. KD < 1 x 10-12M.

[0830] In some embodiments, the antigen-binding molecule described herein binds to human CNX with a KD of 10 pM or less, preferably one of <5 pM, <2 pM, <1 pM, <500 nM, <100 nM, <75 nM, <50 nM, <40 nM, <30 nM, <20 nM, <15 nM, <12.5 nM, <10 nM, <9 nM, <8 nM, <7 nM, <6 nM, <5 nM, <4 nM <3 nM, <2 nM, <1 nM, <500 pM, <400 pM, <300 pM, <200 pM, <100 pM, <50 pM, <40 pM, <30 pM, <20 pM, <10 pM or <1 pM (e.g. as determined by analysis as described in Example 2 herein). In some embodiments, the antigen-binding molecule described herein binds to human CNX with a KD of 100 nM or less, preferably one of <50 nM, <40 nM, <30 nM, <20 nM, <15 nM, <12.5 nM, <10 nM, <9 nM, <8 nM, <7 nM, <6 nM, <5 nM, <4 nM <3 nM, <2 nM, <1 nM, <500 pM, <400 pM, <300 pM, <200 pM, <100 pM, <50 pM, <40 pM, <30 pM, <20 pM, <10 pM or <1 pM (e.g. as determined by analysis as described in Example 2 herein).

[0831] In some embodiments, the antigen-binding molecule described herein binds to human CNX with an ECso of 10 pM or less, preferably one of <5 pM, <2 pM, <1 pM, <500 nM, <100 nM, <75 nM, <50 nM, <40 nM, <30 nM, <20 nM, <15 nM, <12.5 nM, <10 nM, <9 nM, <8 nM, <7 nM, <6 nM, <5 nM, <4 nM <3 nM, <2 nM, <1 nM, <500 pM, <400 pM, <300 pM, <200 pM, <100 pM, <50 pM, <40 pM, <30 pM, <20 pM, <10 pM or <1 pM (e.g. as determined by analysis as described in Example 2 herein).

[0832] In some embodiments, the antigen-binding molecule is cross-reactive for human CNX and a homologue thereof (e.g. mouse CNX). In some embodiments, the antigen-binding molecule is cross-reactive for CNX and CRT. As used herein, a ‘cross-reactive’ antigen-binding molecule / moiety / domain / polypeptide binds to the target antigens for which the antigen-binding molecule / moiety / domain / polypeptide is cross-reactive. For example, an antigen-binding molecule / moiety / domain / polypeptide which is cross-reactive for human CNX and mouse CNX binds to human CNX, and is also capable of binding to mouse CNX. Similarly, an antigen-binding molecule / moiety / domain / polypeptide which is cross-reactive for human CNX and human CRT binds to CNX, and is also capable of binding to CRT. Cross-reactive antigen-binding molecules / moieties / domains / polypeptides may display specific binding to each of the target antigens.

[0833] In some embodiments, the antigen-binding molecule binds to human CNX (e.g. isoform 1 ), and mouse CNX. In some embodiments, the antigen-binding molecule binds to human CNX (e.g. isoform 1 ) and human CRT.

[0834] The antigen-binding molecules of the present disclosure may bind to a particular region of interest of CNX. Antigen-binding molecules according to the present disclosure may bind to linear epitope of CNX, consisting of a contiguous sequence of amino acids ( / .e. an amino acid primary sequence). In some embodiments, an antigen-binding molecules may bind to a conformational epitope of CNX, consisting of a discontinuous sequence of amino acids of the amino acid sequence.

[0835] The region of a given target molecule to which an antigen-binding molecule binds can be determined by the skilled person using various methods well known in the art, including X-ray co-crystallography analysis of antibody-antigen complexes, peptide scanning, mutagenesis mapping, hydrogen-deuterium exchange analysis by mass spectrometry, phage display, competition ELISA and proteolysis-based ‘protection’ methods. Such methods are described, for example, in Gershoni et al., BioDrugs, 2007, 21 (3): 145-156, which is hereby incorporated by reference in its entirety. In preferred embodiments, the region of a peptide / polypeptide to which an antigen-binding molecule binds is determined by hydrogendeuterium exchange analysis by mass spectrometry, performed essentially as described in Example 2 herein.

[0836] In some embodiments, the antigen-binding molecule of the present disclosure binds to a domain of CNX described herein, e.g. the lumenal domain (e.g. lectin domain 1, P domain, lectin domain 2), transmembrane domain or cytoplasmic domain.

[0837] In some embodiments, the antigen-binding molecule of the present disclosure binds to the lumenal domain of CNX. In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:337. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:337.

[0838] In some embodiments, the antigen-binding molecule of the present disclosure binds to the lectin domain of CNX. In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NQ:340. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NQ:340. In some embodiments, the antigen-binding molecule of the present disclosure binds to the P domain of CNX. In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:341. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:341 .

[0839] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:361 . In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:361 . In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:361. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:361 . In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:361 .

[0840] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:362. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:362. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:362. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:362. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:362.

[0841] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:363. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:363. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:363. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:363. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:363.

[0842] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:364. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:364. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:364. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:364. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:364.

[0843] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:365. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:365. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:365. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:365. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:365.

[0844] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:366. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:366. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:366. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:366. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:366.

[0845] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:367. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:367. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:367. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:367. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:367.

[0846] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:368. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:368. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:368. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:368. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:368.

[0847] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:369. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:369. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:369. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:369. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:369.

[0848] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NQ:370. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NQ:370. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NQ:370. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NQ:370. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NQ:370. In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:371 . In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:371 . In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:371 . In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:371 . In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:371 .

[0849] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:372. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:372. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:372. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:372. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:372.

[0850] In some embodiments, the antigen-binding molecule binds to the region of CNX shown in SEQ ID NO:373. In some embodiments, the antigen-binding molecule contacts the region of CNX shown in SEQ ID NO:373. In some embodiments, the antigen-binding molecule binds to CNX via contact with one or more amino acids of the region shown in SEQ ID NO:373. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:373. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:373.

[0851] The ability of an antigen-binding molecule to bind to a given peptide / polypeptide can be analysed by methods well known to the skilled person, including analysis by ELISA, immunoblot (e.g. western blot), immunoprecipitation, surface plasmon resonance and biolayer interferometry.

[0852] In some embodiments, the antigen-binding molecule is capable of binding the same region of CNX, or an overlapping region of CNX, to the region of CNX which is bound by an antibody comprising the VH and VL regions (see e.g. Table C) of one of clones 1 D3, 1 D6, 1 E1 , 1E6, 2C6, 2H6, 3D1, 2G9, 2G12, 2H5, 3F8, 3F9, 4G9, 5A3, 5E8, C001 , C008, C010, C023, C025, C040, C046 and C117. In some embodiments, the antigen-binding molecule is capable of binding the same region of CNX, or an overlapping region of CNX, to the region of CNX which is bound by an antibody comprising the VH and VL regions of C008 or 1 E1 .

[0853] Whether a test antigen-binding molecule binds to the same or an overlapping region of a given target as a reference antigen-binding molecule can be evaluated, for example, by analysis of (i) interaction between the test antigen-binding molecule and the target in the absence of the reference binding molecule, and (ii) interaction between the test antigen-binding molecule in the presence of the reference antigen-binding molecule, or following incubation of the target with the reference antigen-binding molecule. Determination of a reduced level of interaction between the test antigen-binding molecule and the target following analysis according to (ii) as compared to (i) might support an inference that the test and reference antigen-binding molecule bind to the same or an overlapping region of the target. Suitable assays for such analysis include e.g. competition ELISA assays and epitope binning assays.

[0854] In some embodiments, the antigen-binding molecule is an antagonist of CNX, CRT and / or an antagonist of a complex comprising CNX or CRT. In some embodiments, the antigen-binding molecule is capable of inhibiting a function or process mediated by CNX and / or CRT, or mediated by complexes comprising CNX / CRT. In some embodiments, the antigen-binding molecule is capable of inhibiting a function or process mediated by a polypeptide complex comprising CNX or CRT. Herein, ‘inhibition’ refers to a reduction, decrease or lessening relative to a control condition. Suitable assays for investigating the function of CNX and / or CRT, and of complexes comprising CNX / CRT are well known to the skilled person.

[0855] In some embodiments, a complex comprising CNX may be selected from: a CNX:ERp57 complex, a CNX:ERp29 complex and a CNX:CypB complex. In some embodiments, a complex comprising CNX may comprise CNX and a glycopolypeptide. In some embodiments, a complex comprising CRT may be selected from: a CRT:ERp57 complex, a CRT:ERp29 complex and a CRT:CypB complex. In some embodiments, a complex comprising CRT may comprise CRT and a glycopolypeptide.

[0856] In preferred embodiments, a complex comprising CNX is a CNX:ERp57 complex. In preferred embodiments, a complex comprising CRT is a CRT:ERp57 complex.

[0857] Assays for the identification of antigen-binding molecules capable of reducing / inhibiting a function of CNX / CRT and / or of complexes comprising CNX / CRT may comprise treating cells / tissue expressing CNX / CRT and / or a complex comprising CNX / CRT with a test antigen-binding molecule, and subsequently comparing the level of the relevant function to the level observed in an appropriate control condition (e.g. untreated / vehicle-treated / control-treated cells / tissue).

[0858] Antigen-binding molecules capable of reducing / inhibiting a function of CNX / CRT, and / or of a complex comprising CNX / CRT, may be identified using assays comprising detecting the level of a correlate of a function of CNX / CRT, and / or of a complex comprising CNX / CRT, (e.g. the gene and / or protein expression, and / or activity, of one or more proteins whose expression is directly / indirectly upregulated or downregulated as a consequence of a function of CNX / CRT and / or a complex comprising CNX / CRT). Such assays may comprise treating cells / tissue expressing CNX / CRT and / or a complex comprising CNX / CRT with the antigen-binding molecule, and subsequently (e.g. after an appropriate period of time, i.e. a period of time sufficient for the functional consequences of an activity of CNX / CRT and / or a complex comprising CNX / CRT to be observed) comparing the level of the correlate of a function of CNX / CRT, and / or of a complex comprising CNX / CRT, in such cells / tissue to the level of the correlate of the relevant function in an appropriate control condition (e.g. untreated / vehicle-treated / control-treated cells / tissue). In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting a function of CNX / CRT, or of a complex comprising CNX / CRT to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the level of the relevant function observed in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), in a given assay.

[0859] In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring or involving Fc-mediated function. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, independently of Fc-mediated function. That is, in some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, in an Fc region-independent manner.

[0860] The ability of an antigen-binding molecule to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring / involving Fc-mediated function can be evaluated e.g. by analysing the ability of the antigen-binding molecule provided in a format lacking a functional Fc region to inhibit a function of CNX / CRT and / or a complex comprising CNX / CRT. For example, the effect on a function of CNX / CRT, and / or of a complex comprising CNX / CRT, can be investigated using an antigen-binding molecule comprising a ‘silent’ Fc region (e.g. comprising LALA PG substitutions), or using an antigen-binding molecule provided in a format lacking an Fc region (e.g. scFv, Fab etc.).

[0861] In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not involving ADCC. In some embodiments, the antigenbinding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not involving ADCP. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not involving CDC.

[0862] In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding of the antigen-binding molecule to an Fc receptor. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding of the antigen-binding molecule to an Fey receptor. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding of the antigen-binding molecule to one or more of FcyRI, FcyRlla, FcyRllb, FcyRllc, FcyRllla and FcyRI I lb. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding to FcyRllla. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding to FcyRlla. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding to FcyRllb. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding to a complement protein. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring binding to C1q. In some embodiments, the antigen-binding molecule is able to inhibit a function of CNX / CRT, and / or of a complex comprising CNX / CRT, by a mechanism not requiring N297 glycosylation.

[0863] It will be appreciated that in some embodiments, the antigen-binding molecule of the present disclosure achieves is functional effects via a mechanism not involving Fc-mediated function. In some embodiments, the antigen-binding molecule of the present disclosure achieves is functional effects via a mechanism not involving kill i ng / depletion of cells expressing CNX / CRT, or of cells expressing complexes comprising CNX / CRT, e.g. Fc-mediated killing / depletion of such cells.

[0864] In some embodiments, a function of CNX / CRT, or a function of a complex comprising CNX / CRT, may be selected from: extracellular matrix (ECM) degradation, collagen degradation, gelatin degradation, oxireductase activity and disulfide bond reductase activity. A correlate of a function of CNX / CRT, or of a complex comprising CNX / CRT, may e.g. be a product of ECM / collagen / gelatin degradation, or oxireductase / disulfide bond reductase activity.

[0865] In some embodiments, the antigen-binding molecule reduces / inhibits extracellular matrix (ECM) degradation. In some embodiments, the antigen-binding molecule reduces / inhibits collagen degradation. In some embodiments, the antigen-binding molecule reduces / inhibits gelatin degradation. In some embodiments, the antigen-binding molecule reduces / inhibits oxireductase activity. In some embodiments, the antigen-binding molecule reduces / inhibits disulfide bond reductase activity. In some embodiments, the antigen-binding molecule reduces / inhibits ECM degradation mediated by CNX / CRT or a complex comprising CNX / CRT (e.g. a CNX / CRT:ERp57 complex). In some embodiments, the antigen-binding molecule reduces / inhibits collagen degradation mediated by CNX / CRT or a complex comprising CNX / CRT (e.g. a CNX / CRT:ERp57 complex). In some embodiments, the antigen-binding molecule reduces / inhibits gelatin degradation mediated by CNX / CRT or a complex comprising CNX / CRT (e.g. a CNX / CRT:ERp57 complex). In some embodiments, the antigen-binding molecule reduces / inhibits oxireductase activity mediated by CNX / CRT or a complex comprising CNX / CRT (e.g. a CNX / CRT:ERp57 complex). In some embodiments, the antigen-binding molecule reduces / inhibits disulfide bond reductase activity mediated by CNX / CRT or a complex comprising CNX / CRT (e.g. a CNX / CRT:ERp57 complex).

[0866] The ability of an antigen-binding molecule to inhibit ECM / collagen / gelatin degradation can be determined for example by analysis of ECM / collagen / gelatin degradation in the presence of, or following incubation with, the antigen-binding molecule. An antigen-binding molecule which is capable of inhibiting ECM / collagen / gelatin degradation is identified by the observation of a reduction / decrease in the level of ECM / collagen / gelatin degradation in the presence of - or following incubation with - the antigen-binding molecule, as compared to the level of ECM / collagen / gelatin degradation in the absence of the antigenbinding molecule (or in the presence of an appropriate control antigen-binding molecule). Antigen-binding molecules capable of reducing / inhibiting ECM / collagen / gelatin degradation {e.g. by CNX / CRT and / or a complex comprising CNX / CRT) may be identified using assays comprising detecting the level of ECM / collagen / gelatin, or the level of a correlate of ECM / collagen / gelatin degradation (e.g. a product of degraded ECM / collagen / gelatin), e.g. using antibody / reporter-based methods. Collagen / gelatin degradation assays are described e.g. in Hollander, Methods Mol. Biol. (2010) 622:367-78 and Vandooren et al., World J. Biol. Chem. (2011 ) 2(1): 14-24. In preferred embodiments, ECM / collagen / gelatin degradation can be evaluated in an assay performed essentially as described in Example 4 herein.

[0867] For example, a commercial solution of gelatin (2%) can be labeled with 5-Carboxy-X-Rhodamine, Succinimidyl Ester. The labeled gelatin can then be transferred onto sterile coverslips to create a thin layer, and stabilised by glutaraldehyde fixation. A solution of rat tail collagen can be used to coat the coverslips, creating a thin layer of collagen on top of the gelatin. The coverslips can then be transferred in culture vessels and cells with the appropriate degradative activity (e.g. human hepatocellular carcinoma Huh7 cells) can be seeded on the coverslips in the presence of test antigen-binding molecules, and incubated for 48h to allow degradation to occur. The coverslips can then be fixed, and subsequently stained with Hoescht to permit the counting of cells, and then analysed by confocal microscopy. The images acquired can be analysed using Imaged to determine the surface of degraded gelatin and the total area per field. In parallel, the number of nuclei can be calculated and the final result can be normalised to the number of cells in each field.

[0868] For example, a mixture of rat tail collagen and quenched fluorescent DQ collagen type I can be coated and polymerised on the bottom of a 384 well optical grade plate. Cells of the 3t3-vSrc mouse cell line can be seeded on top of the collagen layer in the presence of test antigen-binding molecules, and incubated for 48h to 72h. The fluorescent area of DQ signal from live cells can subsequently be evaluated by high content imaging, and normalised by nucleus count to determine the degraded area / cell.

[0869] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting ECM degradation, collagen degradation or gelatin degradation to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the level of ECM degradation / collagen degradation / gelatin degradation observed in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), in a given assay.

[0870] The ability of an antigen-binding molecule to inhibit oxireductase activity can be determined for example by analysis of oxireductase activity in the presence of, or following incubation with, the antigen-binding molecule. An antigen-binding molecule which is capable of inhibiting oxireductase activity is identified by the observation of a reduction / decrease in the level oxireductase activity in the presence of - or following incubation with - the antigen-binding molecule, as compared to the level of oxireductase activity in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule). Oxireductase activity can be evaluated using any one of a number of methods known to the person skilled in the art. For example, oxireductase activity can be evaluated in an insulin reduction assay, e.g. as described in Hirano et al., Eur J Biochem. (1995) 234(1 ):336-42.

[0871] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting oxireductase activity to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the level of oxireductase activity observed in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), in a given assay.

[0872] The ability of an antigen-binding molecule to inhibit disulfide bond reductase activity can be determined for example by analysis of disulfide bond reductase activity in the presence of, or following incubation with, the antigen-binding molecule. An antigen-binding molecule which is capable of inhibiting disulfide bond reductase activity is identified by the observation of a reduction / decrease in the level disulfide bond reductase activity in the presence of - or following incubation with - the antigen-binding molecule, as compared to the level of disulfide bond reductase activity in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule).

[0873] Disulfide bond reductase activity can be evaluated using any one of a number of methods known to the person skilled in the art. For example, disulfide bond reductase assays may employ antibodies for detecting reduced disulfide bonds in proteins, e.g. antibody clone 0X133, which recognizes polypeptide resident, N-ethylmaleimide (NEM)-modified cysteine residues (see Holbrook et al., Mabs (2016) 8(4): 672-677).

[0874] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting disulfide bond reductase activity to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the level of disulfide bond reductase activity observed in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), in a given assay.

[0875] In some embodiments, an antigen-binding molecule according to the present disclosure may potentiate ( / .e. upregulate, enhance) cell killing of cells comprising / expressing CNX / CRT, or a complex comprising CNX / CRT. In some embodiments, an antigen-binding molecule according to the present disclosure may potentiate ( / .e. upregulate, enhance) cell killing of cancer cells, e.g. cancer cells comprising / expressing CNX / CRT, or a complex comprising CNX / CRT. In some embodiments, an antigen-binding molecule according to the present disclosure may inhibit growth or reduce metastasis of a cancer comprising cells comprising / expressing CNX / CRT, or a complex comprising CNX / CRT. Cell killing can be investigated, for example, using any of the methods reviewed in Zaritskaya et al., Expert Rev Vaccines (2011 ), 9(6):601-616, hereby incorporated by reference in its entirety. Examples of in vitro assays of cytotoxicity / cell killing assays include release assays such as the51Cr release assay, the lactate dehydrogenase (LDH) release assay, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) release assay, and the calcein-acetoxymethyl (calcein-AM) release assay. These assays measure cell killing based on the detection of factors released from lysed cells. Cell killing of a given test cell type by a given effector immune cell type can be analysed e.g. by co-culturing the test cells with the effector immune cells, and measuring the number / proportion of viable / dead (e.g. lysed) test cells after a suitable period of time. Other suitable assays include the xCELLigence real-time cytolytic in vitro potency assay described in Cerignoli et al., PLoS One. (2018) 13(3): e0193498 (hereby incorporated by reference in its entirety). An increase in resistance to cell killing by granzyme B-expressing cells (e.g. effector immune cells), and / or a reduction in susceptibility to cell killing by such cells, relative to a reference level of cell killing (e.g. for that cell type) can be determined by detection of a reduction in the number / proportion of dead (e.g. lysed) test cells, and / or an increase in the number / proportion of live (e.g. viable, non-lysed) test cells, after a given period of time.

[0876] In some embodiments an antigen-binding molecule according to the present disclosure is capable of reducing the number / proportion of cells expressing CNX / CRT, or a complex comprising CNX / CRT. In some embodiments, an antigen-binding molecule according to the present disclosure is capable of depleting / enhancing depletion of such cells.

[0877] In some embodiments, an antigen-binding molecule according to the present disclosure comprises a drug moiety. The antigen-binding moiety may be conjugated to the drug moiety. Antibody-drug conjugates are reviewed e.g. in Parslow et al., Biomedicines. 2016 Sep; 4(3): 14 (hereby incorporated by reference in its entirety). In some embodiments, the drug moiety is or comprises a cytotoxic agent, such that the antigenbinding molecule displays cytotoxicity to a cell expressing CNX / CRT, or a complex comprising CNX / CRT (e.g. a cell expressing CNX / CRT, or a complex comprising CNX / CRT at the cell surface). In some embodiments, the drug moiety is or comprises a chemotherapeutic agent.

[0878] In some embodiments, the antigen-binding molecule of the present disclosure displays anticancer activity. In some embodiments, the antigen-binding molecule of the present disclosure increases killing of cancer cells. In some embodiments, the antigen-binding molecule of the present disclosure causes a reduction in the number of cancer cells in vivo, e.g. as compared to an appropriate control condition. The cancer may be a cancer expressing CNX / CRT, or a complex comprising CNX / CRT.

[0879] In some embodiments, an antigen-binding molecule according to the present disclosure reduces / inhibits growth of a cancer and / or of a tumor of a cancer. In some embodiments, an antigen-binding molecule reduces tissue invasion by cells of a cancer. In some embodiments, an antigen-binding molecule reduces metastasis of a cancer. In some embodiments, the antigen-binding molecule displays anticancer activity. In some embodiments, the antigen-binding molecule reduces the growth / proliferation of cancer cells. In some embodiments, the antigen-binding molecule reduces the survival of cancer cells. In some embodiments, the antigen-binding molecule increases the killing of cancer cells. In some embodiments, the antigen-binding molecule of the present disclosure causes a reduction in the number of cancer cells e.g. in vivo. The cancer may be a cancer comprising cells expressing CNX and / or CRT.

[0880] The antigen-binding molecule of the present disclosure may be analysed for the properties described in the preceding paragraph in appropriate assays. Such assays include e.g. in vivo models, e.g. performed essentially as described in Example 5 herein.

[0881] In some embodiments, administration of an antigen-binding molecule according to the present disclosure may cause one or more of: inhibition of the development / progression of the cancer, a delay to / prevention of onset of the cancer, a reduction in / delay to / prevention of tumor growth, a reduction in / delay to / prevention of tissue invasion, a reduction in / delay to / prevention of metastasis, a reduction in the severity of the symptoms of the cancer, a reduction in the number of cancer cells, a reduction in tumour size / volume, and / or an increase in survival (e.g. progression free survival or overall survival), e.g. as determined in an appropriate model.

[0882] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting tumor growth (e.g. in an in vivo model, e.g. of liver cancer) to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the tumor growth observed in the absence of treatment with the antigen-binding molecule (or following treatment with an appropriate control antigenbinding molecule known not to influence tumor growth), in a given assay.

[0883] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting metastasis (e.g. in an in vivo model, e.g. of metastasis of breast cancer to the lung) to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the level of metastasis observed in the absence of treatment with the antigen-binding molecule (or following treatment with an appropriate control antigen-binding molecule known not to influence metastasis), in a given assay.

[0884] In some embodiments, the antigen-binding molecule of the present disclosure is capable of increasing survival of subjects having a cancer (e.g. in an in vivo model, e.g. of liver cancer or breast cancer) to more than 1 times, e.g. one of >1.01 times, >1.02 times, >1.03 times, >1.04 times, >1.05 times, >1.1 times, >1 .2 times, >1 .3 times, >1 .4 times, >1 .5 times, >1 .6 times, >1 .7 times, >1 .8 times, >1 .9 times, >2 times, >3 times, >4 times, >5 times, >6 times, >7 times, >8 times, >9 times or >10 times the level of survival observed in the absence of treatment with the antigen-binding molecule (or following treatment with an appropriate control antigen-binding molecule known not to influence survival), in a given assay.

[0885] In some embodiments, an antigen-binding molecule according to the present disclosure reduces / inhibits pathology of a disease / condition characterised by ECM degradation in a subject. In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting pathology of a disease / condition characterised by ECM degradation to less than 1 times, e.g. <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times, <0.1 times, <0.05 times, or <0.01 times the level of observed in the absence of treatment with the antigen-binding molecule (or following treatment with an appropriate control antigenbinding molecule known not to influence pathology of the disease / condition), in a given assay.

[0886] In some embodiments, the antigen-binding molecule is capable of being internalised by cells expressing CNX / CRT. In some embodiments, the antigen-binding molecule is capable of being internalised into a given cell in a CNX-dependent manner. The antigen-binding molecule may be internalised by clatherin- mediated endocytosis.

[0887] Assays for the identification of antigen-binding molecules capable of being internalised by cells expressing CNX / CRT may comprise treating cells / tissue expressing CNX / CRT with a test antigen-binding molecule, and subsequently comparing the level of internalisation to the level observed in an appropriate control condition (e.g. cells / tissue not expressing CNX / CRT e.g. a version of a cell / tissue in which CNX / CRT expression has been knocked-out).

[0888] Antigen-binding molecules capable of being internalised by cells expressing CNX / CRT may be identified by assays comprising detecting the level of a test antigen-binding molecule in a cell following exposure of said cell to the test antigen-binding molecule. Such assays may comprise treating cells / tissue with the antigen-binding molecule, and subsequently (e.g. after an appropriate period of time, i.e. a period of time sufficient for endocytosis to be observed) comparing the level of antigen-binding molecule in the cells to the level of the antigen-binding molecule in cells of an appropriate control condition (e.g. cells / tissue not expressing CNX / CRT).

[0889] Antibody-binding molecule internalisation can be evaluated in an assay performed essentially as described in Example 5 herein.

[0890] In some embodiments, the antigen-binding molecule of the present disclosure is internalised by a cell expressing CNX / CRT at a level which is greater than 1 times, e.g. one of >1.01 times, >1.02 times, >1.03 times, >1.04 times, >1.05 times, >1.1 times, >1.2 times, >1.3 times, >1.4 times, >1.5 times, >1.6 times, >1 .7 times, >1 .8 times, >1 .9 times, >2 times, >3 times, >4 times, >5 times, >6 times, >7 times, >8 times, >9 times or >10 times the level internalised at the same time point by a control cell (e.g. a cell of the same cell type with CNX / CRT expression knocked out). In some embodiments, evaluation of the antigenbinding molecule internalisation for the purposes of such comparison is performed after more than 1 hour, e.g. one of >2 hours, >3 hours, >4 hours, >5 hours, >6 hours, >7 hours, >8 hours, >9 hours, >10 hours, >11 hours, >12 hours, >13 hours, >14 hours, >15 hours, >16 hours, >17 hours, >18 hours, >19 hours, >20 hours, >21 hours, >22 hours, >23 hours or >24 hours following incubation of the antigen-binding molecule with the cells. Nucleic acids and vectors

[0891] The present disclosure provides a nucleic acid, or a plurality of nucleic acids, encoding an antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex or a constituent polypeptide thereof according to the present disclosure. In some embodiments, the nucleic acid(s) comprise or consist of DNA and / or RNA.

[0892] In some embodiments, the nucleic acid(s) may be, or may be comprised in, a vector, or a plurality of vectors. That is, the nucleotide sequence(s) of the nucleic acid(s) may be contained in vector(s). The antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex or a constituent polypeptide thereof according to the present disclosure may be produced within a cell by transcription from a vector encoding the antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex or a constituent polypeptide thereof and subsequent translation of the transcribed RNA.

[0893] Accordingly, the present disclosure also provides a vector, or plurality of vectors, comprising the nucleic acid or plurality of nucleic acids according to the present disclosure. The vector may facilitate delivery of the nucleic acid(s) encoding an antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex or a constituent polypeptide thereof according to the present disclosure to a cell. The vector may be an expression vector comprising elements required for expressing nucleic acid(s) comprising / encoding an antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex or a constituent polypeptide thereof according to the present disclosure. The vector may comprise elements facilitating integration of the nucleic acid(s) into the genomic DNA of cell into which the vector is introduced.

[0894] Nucleic acids and vectors according to the present disclosure may be provided in purified or isolated form, i.e. from other nucleic acid, or naturally-occurring biological material.

[0895] A ‘vector’ as used herein is a nucleic acid molecule used as a vehicle to transfer exogenous nucleic acid into a cell. The vector may be a vector for expression of the nucleic acid in the cell. Such vectors may include a promoter sequence operably linked to the nucleotide sequence encoding the sequence to be expressed. A vector may also include a termination codon and expression enhancers. Any suitable vectors, promoters, enhancers and termination codons known in the art may be used to express a peptide or polypeptide from a vector according to the present disclosure.

[0896] The term ‘operably linked’ may include the situation where a selected nucleic acid sequence and regulatory nucleic acid sequence (e.g. promoter and / or enhancer) are covalently linked in such a way as to place the expression of nucleic acid sequence under the influence or control of the regulatory sequence (thereby forming an expression cassette). Thus a regulatory sequence is operably linked to the selected nucleic acid sequence if the regulatory sequence is capable of effecting transcription of the nucleic acid sequence. The resulting transcript(s) may then be translated into a desired peptide(s) / polypeptide(s). Vectors contemplated in connection with the present disclosure include DNA vectors, RNA vectors, plasmids (e.g. conjugative plasmids (e.g. F plasmids), non-conjugative plasmids, R plasmids, col plasmids, episomes), viral vectors (e.g. retroviral vectors, e.g. gammaretroviral vectors (e.g. murine Leukemia virus (MLV)-derived vectors, e.g. SFG vector), lentiviral vectors, adenovirus vectors, adeno- associated virus vectors, vaccinia virus vectors and herpesvirus vectors), transposon-based vectors, and artificial chromosomes (e.g. yeast artificial chromosomes), e.g. as described in Maus et al., Annu Rev Immunol (2014) 32:189-225 and Morgan and Boyerinas, Biomedicines (2016) 4:9, which are both hereby incorporated by reference in their entirety.

[0897] In some embodiments, the vector may be a eukaryotic vector, e.g. a vector comprising the elements necessary for expression of protein from the vector in a eukaryotic cell. In some embodiments, the vector may be a mammalian vector, e.g. comprising a cytomegalovirus (CMV) or SV40 promoter to drive protein expression.

[0898] Constituent polypeptides of an antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex according to the present disclosure may be encoded by different nucleic acids of the plurality of nucleic acids, or by different vectors of the plurality of vectors.

[0899] Cells comprising / expressing the antigen-binding molecules / moieties and their constituent polypeptides

[0900] The present disclosure also provides a cell comprising or expressing an antigen-binding molecule / antigen-binding moiety / antigen-binding polypeptide complex or a constituent polypeptide thereof according to the present disclosure. Also provided is a cell comprising or expressing a nucleic acid, a plurality of nucleic acids, a vector or a plurality of vectors according to the present disclosure.

[0901] The cell may be a eukaryotic cell, e.g. a mammalian cell. The mammal may be a primate (rhesus, cynomolgous, non-human primate or human) or a non-human mammal (e.g. rabbit, guinea pig, rat, mouse or other rodent (including any animal in the order Rodentia), cat, dog, pig, sheep, goat, cattle (including cows, e.g. dairy cows, or any animal in the order Bos), horse (including any animal in the order Equidae), donkey, and non-human primate).

[0902] In some embodiments, the cell is, or is derived from, a cell type commonly used for the expression of polypeptides for use in therapy in humans. Exemplary cells are described e.g. in Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100:3451-3461 (hereby incorporated by reference in its entirety), and include e.g. CHO, HEK 293, PER.C6, NS0 and BHK cells. In preferred embodiments, the cell is, or is derived from, a CHO cell.

[0903] The present disclosure also provides a method for producing a cell comprising a nucleic acid(s) or vector(s) according to the present disclosure, comprising introducing a nucleic acid, a plurality of nucleic acids, a vector or a plurality of vectors according to the present disclosure into a cell. In some embodiments, introducing an isolated nucleic acid(s) or vector(s) according to the present disclosure into a cell comprises transformation, transfection, electroporation or transduction (e.g. retroviral transduction). The present disclosure also provides a method for producing a cell expressing / comprising an antigenbinding molecule / antigen-binding moiety or a constituent polypeptide thereof according to the present disclosure, comprising introducing a nucleic acid, a plurality of nucleic acids, a vector or a plurality of vectors according to the present disclosure in a cell. In some embodiments, the methods additionally comprise culturing the cell under conditions suitable for expression of the nucleic acid(s) or vector(s) by the cell. In some embodiments, the methods are performed in vitro.

[0904] The present disclosure also provides cells obtained or obtainable by the methods according to the present disclosure.

[0905] Producing the antigen-binding molecules

[0906] Antigen-binding molecules and polypeptides according to the present disclosure may be prepared according to methods for the production of antibody-drug conjugates known to the skilled person.

[0907] Antigen-binding molecules and components thereof (e.g. antigen-binding moieties / polypeptides) according to the present disclosure may be prepared by chemical synthesis, e.g. liquid or solid phase synthesis. For example, peptides / polypeptides can be synthesised using the methods described in, for example, Chandrudu et al., Molecules (2013), 18: 4373-4388, which is hereby incorporated by reference in its entirety.

[0908] Alternatively, antigen-binding molecules and components thereof (e.g. antigen-binding moieties / polypeptides) may be produced by recombinant expression. Molecular biology techniques suitable for recombinant production of polypeptides are well known in the art, such as those set out in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th Edition), Cold Spring Harbor Press, 2012, and in Nat Methods. (2008); 5(2): 135-146 both of which are hereby incorporated by reference in their entirety. Methods for the recombinant production of antigen-binding moieties / polypeptides are also described in Frenzel et al., Front Immunol. (2013); 4: 217 and Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100: 3451-3461 , both of which are hereby incorporated by reference in their entirety.

[0909] In some cases, the antigen-binding molecules of the present disclosure and components thereof (e.g. antigen-binding moieties are comprised of more than one polypeptide chain. In such cases, production of the antigen-binding molecule / moiety may comprise transcription and translation of more than one polypeptide, and subsequent association of the polypeptide chains to form the antigen-binding molecule / moiety.

[0910] For recombinant production according to the present disclosure, any cell suitable for the expression of polypeptides may be used. The cell may be a prokaryote or eukaryote. In some embodiments, the cell is a prokaryotic cell, such as a cell of archaea or bacteria. In some embodiments, the bacteria may be Gram-negative bacteria such as bacteria of the family Enterobacteriaceae, for example Escherichia coll. In some embodiments, the cell is a eukaryotic cell such as a yeast cell, a plant cell, insect cell or a mammalian cell, e.g. a cell described hereinabove. In some cases, the cell is not a prokaryotic cell because some prokaryotic cells do not allow for the same folding or post-translational modifications as eukaryotic cells. In addition, very high expression levels are possible in eukaryotes and proteins can be easier to purify from eukaryotes using appropriate tags. Specific plasmids may also be utilised which enhance secretion of the protein into the media.

[0911] In some embodiments polypeptides may be prepared by cell-free-protein synthesis (CFPS), e.g. according to a system described in Zemella et al. Chembiochem (2015) 16(17): 2420-2431, which is hereby incorporated by reference in its entirety.

[0912] Production of antigen-binding molecules and components thereof (e.g. antigen-binding moieties / polypeptides) may involve culture or fermentation of a eukaryotic cell modified to express the polypeptide(s) of interest. The culture or fermentation may be performed in a bioreactor provided with an appropriate supply of nutrients, air / oxygen and / or growth factors. Secreted proteins can be collected by partitioning culture media / fermentation broth from the cells, extracting the protein content, and separating individual proteins to isolate secreted polypeptide(s). Culture, fermentation and separation techniques are well known to those of skill in the art, and are described, for example, in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th Edition; incorporated by reference herein above).

[0913] Bioreactors include one or more vessels in which cells may be cultured. Culture in the bioreactor may occur continuously, with a continuous flow of reactants into, and a continuous flow of cultured cells from, the reactor. Alternatively, the culture may occur in batches. The bioreactor monitors and controls environmental conditions such as pH, oxygen, flow rates into and out of, and agitation within the vessel such that optimum conditions are provided for the cells being cultured.

[0914] Following culturing the cells that express the polypeptide(s), the polypeptide(s) of interest may be isolated. Any suitable method for separating proteins from cells known in the art may be used. In order to isolate the polypeptide, it may be necessary to separate the cells from nutrient medium. If the polypeptide(s) are secreted from the cells, the cells may be separated by centrifugation from the culture media that contains the secreted polypeptide(s) of interest. If the polypeptide(s) of interest collect within the cell, protein isolation may comprise centrifugation to separate cells from cell culture medium, treatment of the cell pellet with a lysis buffer, and cell disruption e.g. by sonification, rapid freeze-thaw or osmotic lysis.

[0915] It may then be desirable to isolate the polypeptide(s) of interest from the supernatant or culture medium, which may contain other protein and non-protein components. A common approach to separating protein components from a supernatant or culture medium is by precipitation. Proteins of different solubilities are precipitated at different concentrations of precipitating agent such as ammonium sulfate. For example, at low concentrations of precipitating agent, water soluble proteins are extracted. Thus, by adding different increasing concentrations of precipitating agent, proteins of different solubilities may be distinguished. Dialysis may be subsequently used to remove ammonium sulfate from the separated proteins. Other methods for distinguishing different proteins are known in the art, for example ion exchange chromatography and size chromatography. These may be used as an alternative to precipitation or may be performed subsequently to precipitation.

[0916] Once the polypeptide(s) of interest have been isolated from culture it may be desired or necessary to concentrate the polypeptide(s). A number of methods for concentrating proteins are known in the art, such as ultrafiltration or lyophilisation.

[0917] Antigen-binding moieties / polypeptides / polypeptide complexes according to the present disclosure may be conjugated to linker-payload moieties according to the present disclosure for the production of antigenbinding molecules according to the present disclosure by any suitable techniques, which are well known to the skilled person and routinely employed in the art.

[0918] General methods for the conjugation of antigen-binding moieties / polypeptides / polypeptide complexes to linker-payload moieties are described e.g. in Chudasama et al., Nature Chemistry, (2016), 8:114-119, Baah et al., Molecules. (2021) 26(10): 2943, and Walsh et al., Chem. Soc. Rev. (2021 ) 50:1305-1353, all of which are hereby incorporated by reference in their entirety. Conjugation of antigen-binding moieties and linker-payload moieties and purification of antigen-binding molecules produced by such conjugation is described e.g. in Beck et al., (2017) Nat Rev Drug Discov 16: 315-337; Peters and Brown Biosci Rep (2015) 35: art:e00225; McCombs and Owen, The AAPS Journal (2015) 17: 339-351; Jackson, Org Process Res Dev (2016) 20: 852-866; and Olivier and Hurvitz, Antibody-Drug Conjugates: Fundamentals, Drug Development, and Clinical Outcome to Target Cancer (2016) Wiley.

[0919] Antigen-binding moieties according to the present disclosure may be conjugated to linker-payload moieties according to the present disclosure by any suitable techniques, which are well known to the skilled person and routinely employed in the art. Such methods are described e.g. in Chudasama et al., Nature Chemistry, (2016), 8:114-119, Baah et al., Molecules. (2021 ) 26(10): 2943, and Walsh et al., Chem. Soc. Rev. (2021 ) 50:1305-1353, all of which are hereby incorporated by reference in their entirety.

[0920] Conjugation of antigen-binding moieties and linker-payload moieties and the purification of antigenbinding molecules produced by such conjugation can be performed e.g. as described in in Beck et al., (2017) Nat Rev Drug Discov 16: 315-337; Peters and Brown Biosci Rep (2015) 35: art:e00225; McCombs and Owen, The AAPS Journal (2015) 17: 339-351; Jackson, Org Process Res Dev (2016) 20: 852-866; or Olivier and Hurvitz, Antibody-Drug Conjugates: Fundamentals, Drug Development, and Clinical Outcome to Target Cancer, (2016) Wiley, all of which are hereby incorporated by reference in their entirety. Other relevant disclosures relating to conjugation and linkers are: Tsuchikama and An, Protein Cell. (2018) 9(1): 33-46, Khongorzul et al., Mol Cancer Res (2020) 18 (1 ): 3-19 and Drago et al., Nature Reviews Clinical Oncology (2021) 18: 327-344, all of which are hereby incorporated by reference in their entirety.

[0921] Lysine amide coupling Lysine-based conjugation is one of the most widely used non-specific conjugation strategies. Such conjugation occurs on reactive amine side chains of lysine residues due to their good nucleophilicity. Immunoglobulin scaffolds contains over 80 lysine residues, most of which are exposed on the surface of the molecule. Among the surface lysine residues, more than 20 have been shown as highly solvent- accessible and can serve as potential ADC conjugation sites. Lysine conjugation follows two main strategies that result in the formation of a stable amide or amidine bond between the protein and the drug-linker complex. Generally speaking, activated esters on the drug-linker complexes, often O- succinimide reagents such as N-hydroxysuccinimidyl (NHS) or sulfo-NHS esters, react with the antibody lysine residues and achieve the conjugation via amide bonds. On the other hand, stable amidine bonds can be generated on an antibody by the reaction of imido ester compounds, such as Traut’s reagent, with antibody lysine residues.

[0922] A one-step conjugation of a drug-linker moiety containing an amine-reactive group to the antibody via amide bonds is known, as well as two-step conjugation, where in the first step, a small bi-functional reagent containing both an amine- and a thiol-reactive functional groups is reacted with the available lysine e-amino groups to serve as a chemical adaptor, leaving free thiol-reactive groups on the antibody. In the second step, the payload drugs or drug-linker complexes are attached to the thiol-reactive groups introduced previously to form the ADC. The two-step approach is often used when the drug / drug-linker complex contains a thiol-reactive module or as an alternative route when introducing an amine-reactive module into the drug or drug-linker complex is proven to be difficult. Four small adaptors commonly used in the two-step conjugation: SPDB disulfide, MCC (maleimidomethyl cyclohexane-1-carboxylate), sulfo- SPDB, and Hydrazine.

[0923] Cysteine coupling

[0924] Cysteine modification occurs most commonly by 1 ,4-conjugate addition to A / -substituted maleimides. Maleimides are particularly attractive reagents due to their synthetic accessibility and rapid reaction rates with cysteine under mild conditions. The resulting thiosuccinimide conjugates are inherently unstable, due to their propensity towards retro-Michael addition. This instability can be mitigated by forcing postconjugation hydrolysis of the thiosuccinimide, creating a stable chemical linkage. Accordingly, a number of “self-hydrolysing” maleimides have now been developed, with ring-opening catalysed by adjacent functional groups such as primary amine, polyethylene glycol (PEG) and A / -aryl amongst the most promising. Other reagents including a-halocarbonyls, palladium oxidative-addition complexes, https: / / pubs.rsc.orQ / en / content / articlelandinq / 2021 / cs / d0cs00310Q ethynylphosphonamidates, vinylphosphonites and ethynylbenziodoxolones.

[0925] Some non-maleimide cysteine conjugations are summarised in Kang, et al., Chem Sci (2021 ) 12, 13613- 13647 (doi: 10.1039 / D1 SC02973H), and include the use of:

[0926] (i) alkynyl carboxylic acid derivatives;

[0927] (ii) 5-methylene pyrrolone (5MP);

[0928] (iii) 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB);

[0929] (iv) phenyloxadiazole sulfone (PODS);

[0930] (v) aza-dibenzocyclooctyne (DBCO); (vi) phosphonamidite;

[0931] (vii) 3-arylpropionitrile (APN);

[0932] (viii) perfluoroarene;

[0933] (ix) ethynylbenziodoxolone (EBX);

[0934] (x) bicyclo[1.1.0]butane (BCB) carboxylic amide; and

[0935] (xi) allenamide.

[0936] Another possible approach is described in Cheng, et al., Front. Oncol. 12:951589 (doi: 10.3389 / fonc.2022.951589) where 2-methylsulfonyl pyrimidine is used instead of a maleimide.

[0937] Genetic modification of the number of accessible cysteine residues on an antibody surface is a method to achieve site-selective and homogeneous modification. For example, in THIOMABs, the engineered cysteine is installed on an anti-MUC16 antibody by mutation of heavy chain alanine 114 (HC-A114). Other approaches have engineered antibodies to contain cysteine mutations at D265C, S239C, E269C, K326C or A327C, or to insert additional cysteines before and after positions HC-S239, HC-A114, and LC- V205.

[0938] A further approach, uses cysteine / disulphide rebridging to produce ADCs with highly homogenous drug- to-antibody ratios (DAR) at native interchain disulfides. Cycteine / disulphide rebridging methods for ADC production include ThioBridge®. Briefly, in the ThioBridge® approach, interchain disulphide bonds in the antigen-binding moiety (e.g. an antibody or antigen-binding fragment thereof) are reduced to release sulfhydryl residues for the conjugation. Conjugation then occurs with a linker-payload moiety which comprises a bis-functional electrophilic group capable of reacting with a pair of sulfhydryl residues on the antigen-binding moiety to covalently rebridge the disulphide bonds. Cysteine / disulphide rebridging approaches are reviewed in Antibody-Drug Conjugates: Methods and Protocols (Humana Press, 2020), at Chapter 8: Bridged Cysteine Conjugations (Bird et al.), which is hereby incorporated by reference in its entirety.

[0939] Non-natural amino acid incorporation by genetic engineering

[0940] Site-specific incorporation of non-canonical amino acids (ncAAs) into antibodies results in an efficient approach to the site-specific modification of antibodies, and therefore homogeneous ADCs.

[0941] NcAAs bearing unique functionalities, such as ketones, azides, cyclopropenes or diene functional groups, have been developed and incorporated into antibodies. Such ncAAs include p-acetylphenylalanine (pAcF), which has a ketone side chain which can participate in oxime ligation reactions; Ne-(1- methylcycloprop-2-enecarboxamido)-lysine (CpK), which has a cyclopropene side chain which can participate in IEDDA reactions; para-azidomethyl phenylalanine (pAMF), which has a an azide side chain which can undergo click reactions; spiro[2.4]hepta-4,6-diene-lysine (SCpHK), which has a spiro[2.4]hepta-4,6-diene side chain which can participate in Diels-Alder reactions; and N6-(2- azidoethoxy)-carbonyl-L-lysine (AzK), which has a an azide side chain which can undergo click reactions.

[0942] Azide-containing ncAAs can undergo rapid CuAAC or SPAAC reactions under physiological conditions, para-azidophenylalanine (pAzF) can undergo reactions with, for example, cyclooctyne-functionalised linkers and dibenzylcyclooctyne (DBCO)-functionalised linkers. A cyclopropene derivative of lysine (N c-[((2-methylcycloprop-2-en-1-yl)methoxy)carbonyl]-l-lysine; CypK) can undergo a rapid and efficient inverse-electron demand Diels-Alder (IEDDA) reaction with a tetrazine-functionalised linker.

[0943] Cyclopentadiene-containing ncAAs, spiro[2.4]hepta-4,6-diene-lysine (SCpHK) and cyclopentadiene-lysine (CpHK), can undergo irreversible Diels-Alder cycloadditions with maleimide-modified drugs.

[0944] Self-labelling protein tag coupling

[0945] Self-labelling protein tags are able to react with a specific partner ligand to form a covalent bond between the tag and the partner ligand. The self-labelling protein tag can be expressed as a fusion protein with a protein / polypeptide of interest. A linker-payload moiety may comprise a specific partner ligand to a given self-labelling protein. Thus self-labelling protein tags allow the covalent attachment of a fusion protein (e.g. an antigen-binding moiety:self-labelling protein tag fusion protein) with a specific partner ligand (e.g. a linker-payload moiety comprising the specific partner ligand).

[0946] Self-labelling proteins are known in the art, for example, SNAP-tag, CLIP-tag and HaloTag. Self-labelling proteinJigand pairings include e.g. SNAP-tag and O6-benzylguanine or derivatives thereof, CLIP-tag and benzylcytosine or derivatives thereof, HaloTag and chloroalkane or derivatives thereof. Self-labelling proteins are described in Wilhelm et al., 2021 Biochemistry 60(33): 2560-2575.

[0947] In some embodiments, the self-labelling protein tag is a SNAP-tag. The SNAP-tag is a modified form of the 20-kDa monomeric DNA repair enzyme, human O6-alkylguanine-DNA-alkyltransferase (AGT), The SNAP-tag can be specifically conjugated with synthetic O6-benzylguanine (BG) derivatives, resulting in a stable thioether bond between a reactive cysteine residue in the SNAP-tag and the BG derivative. The SNAP-tag can be appended onto the N- or C-terminus of proteins without affecting the function of a large number of fusion proteins, for example, in this approach the antigen-binding moiety may be expressed as a fusion protein with the SNAP-tag. The fusion protein may then be exposed to a BG-modified payload moiety to allow for conjugation. SNAP-tag technology is described in Hussain, et al., Nature Protocols 14:3101-3125 (2019). A SNAP-tag may comprise, or consist of, an amino acid sequence having at least 60% amino acid sequence identity, e.g. one of >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100% amino acid sequence identity to SEQ ID NO:419.

[0948] Enzymatic conjugation

[0949] Enzymes can be used to achieve site-selective antibody modification due to their high specificity and mild reaction conditions. Enzymes can either directly attach a payload to a specific amino acid sequence or introduce a reactive functionality on the antibody that can be further functionalised with the desired payload.

[0950] Transpeptidation using sortase

[0951] Sortase-mediated antibody conjugation (SMAC) technology is an additional enzymatic ligation approach. SMAC-technology uses S. aureus sortase A, which is a transpeptidase that cleaves the amide bond between threonine and glycine residues in the LPXTG (X = any amino acid) pentapeptide motif, and subsequently catalyses the attachment of glycine-functionalised payloads to the newly generated C- terminus. The sortase recognition motif and a Strep II tag, which is used to aid removal of unreacted antibody, were fused to the light and heavy chain C-terminus of different antibodies. Sortase-mediated conjugation can then be used to attach a series of penta-glycine tagged payloads.

[0952] Transpeptidation using microbial transglutaminase

[0953] The use of bacterial transglutaminases is a powerful approach for site-specific incorporation of the payload into the antibody. A transglutaminase derived from Streptomyces mobaraensis catalyzes transpeptidation where a primary amine-containing linker is covalently attached to the primary amide side chain of a specific glutamine (Q295) within deglycosylated antibodies, resulting in ADCs with a defined DAR arising from the conjugation of 2 linker-payloads (one conjugation site per heavy chain). An N297Q mutation prior to this conjugation provides two more reaction sites (resulting in the conjugation of 4-linker- payloads). An alternative version using a peptide sequence-specific transglutaminase. This enzyme recognizes and utilizes LLQG motif that is genetically incorporated, resulting in site-specific antibody-drug conjugation. Another advantage of this LLQG-specific bacterial transglutaminase is that conjugation sites can be flexibly laid by inserting this short peptide motif within the antibody structure. Further alternative approaches allow for the use of transglutaminase without deglycosylation.

[0954] N-Glycan engineering

[0955] Asn297 (N297) within the Fc domain and the N-glycan on this residue are conserved in all IgG classes, making these components attractive reaction sites for broadly applicable ADC conjugation. Incorporation of an aldehyde group on the N-glycan terminus using p-1 ,4-galactosyltransferase (GalT) and a-2,6- sialyltransferase (SialT) introduce a sialic acid on each N-glycan terminus, which is subsequently converted into an aldehyde group using NalO4 under mild oxidation conditions. The aldehyde groups generated can then used to conjugate aminooxyfunctionalized payloads.

[0956] Another approach is to incorporate non-natural saccharides possessing orthogonal reaction handles into the antibody. A technology based on this strategy is the GlycoConnect in which the glycan chain at Asn297 is trimmed using the endoglycosidase Endo S2 and then azide groups are introduced using a mutant galactosyl transferase GalT(Y289L) and N-azidoacetylgalactosamine (GalNAz). The azide handles can be used for a strain-promoted click reaction with payloads.

[0957] In some embodiments, the linker-payload terminates in an amino group which is conjugated to the antigen-binding molecule using transglutaminase.

[0958] In some embodiments, the method further comprises purifying / isolating the antigen-binding molecule ( / .e. from unreacted precursors and / or by-products). In some embodiments, the antigen-binding molecule may be purified / isolated by chromatography, e.g. size-exclusion chromatography.

[0959] The present disclosure also provides an antigen-binding molecule obtained or obtainable by the methods of the present disclosure. Particular exemplary antigen-binding molecules

[0960] The present disclosure provides antigen-binding molecules comprising an antigen-binding moiety (e.g. a CNX-binding moiety), optionally further comprising a linker-payload moiety.

[0961] In some embodiments, the antigen-binding moiety comprises:

[0962] (a)

[0963] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0964] HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and

[0965] (II) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; or

[0966] (b)

[0967] (I) a heavy chain variable (VH) region incorporating the following CDRs:

[0968] HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and

[0969] (II) a light chain variable (VL) region incorporating the following CDRs:

[0970] LC-CDR1 having the amino acid sequence of SEQ ID NO:179 LC-CDR2 having the amino acid sequence of SEQ ID NQ:180 LC-CDR3 having the amino acid sequence of SEQ ID NO:173.

[0971] In some embodiments, the antigen-binding moiety comprises:

[0972] (ii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:40; or

[0973] (I) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:165; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:178.

[0974] In some embodiments, the antigen-binding moiety comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 279 or 280 In some embodiments, the antigen-binding moiety comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 281. In some embodiments, the antigen-binding moiety comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 418. In some embodiments, the antigen-binding moiety further comprises a SNAP protein. In some embodiments the SNAP protein comprises or consists of an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 419. In some embodiments, the antigen-binding moiety comprises or consists of an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 417.

[0975] In some embodiments, the antigen-binding molecule comprises a linker-payload moiety. As used herein, a linker-payload moiety refers to a moiety comprising one or more payload moieties, and a linker moiety for linking the payload moiety(Zies) to the antigen-binding moiety of the antigen-binding molecule.

[0976] In some embodiments, the payload moiety comprises or consists of a microtubule-targeting agent, e.g. an auristatin. In some embodiments, the payload moiety comprises or consists of monomethyl auristatin E (MMAE) or auristatin F.

[0977] In some embodiments, the payload moiety comprises or consists of a DNA-targeting agent, e.g. a topoisomerase inhibitor, e.g. a TOP1 inhibitor. In some embodiments, the payload moiety comprises or consists of camptothecin or a derivative thereof. In some embodiments, the payload moiety comprises or consists of deruxtecan (Dxd).

[0978] In some embodiments, the linker moiety comprises a maleimidocaproyl (MC) group. In some embodiments, the linker moiety further comprises a cathepsin-cleavable group, e.g. a dipeptide (e.g. valine-citruline (Val-Cit or VC) linker) or a tetrapeptide (e.g. a glycine-glycine-phenylalanine-glycine (GGFG) linker). In some embodiments, the linker moiety further comprises a spacer moiety, e.g. a paraaminobenzyl (PAB).

[0979] In some embodiments, the linker moiety is MC-Val-Cit-PAB. In some embodiments, the linker moiety is MC-GGFG.

[0980] In some embodiments, the linker-payload moiety is MC-Val-Cit-PAB-MMAE. In some embodiments, the linker-payload component of the antigen-binding molecule has the following structure:

[0981] In some embodiments, the linker-payload moiety is MC-GGFG-Dxd. In some embodiments, the linkerpayload component of the antigen-binding molecule has the following structure:

[0982]

[0983] In some embodiments, the antigen-binding molecule has the following structure (1 E1-MC-Val-Cit-PAB- MMAE): wherein ABM is an antigen-binding moiety which comprises:

[0984] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0985] HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and

[0986] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0987] LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; optionally, wherein the antigen-binding moiety comprises a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:40; further optionally wherein the antigen-binding moiety is an IgG monoclonal antibody.

[0988] In some embodiments, the antigen-binding molecule is 1 E1-MC-Val-Cit-PAB-MMAE and the DAR is 4.

[0989] In some embodiments, the antigen-binding molecule has the following structure (1 E1-MC-GGFG-Dxd):

[0990] wherein ABM is an antigen-binding moiety which comprises:

[0991] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0992] HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and

[0993] (ii) a light chain variable (VL) region incorporating the following CDRs:

[0994] LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; optionally wherein the antigen-binding moiety comprises a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:40; further optionally wherein the antigen-binding moiety is an IgG monoclonal antibody.

[0995] In some embodiments, the antigen-binding molecule is 1 E1-MC-GGFG-Dxd and the DAR is 8.

[0996] In some embodiments, the linker-payload moiety is benzyl-guanine Auristatin F. In some embodiments, the linker-payload component of the antigen-binding molecule has the following structure:

[0997] In some embodiments, the antigen-binding molecule has the following structure (scFV(C008)-SNAP- Auristatin F): wherein ABM is an antigen-binding moiety which comprises:

[0998] (i) a heavy chain variable (VH) region incorporating the following CDRs:

[0999] HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and

[1000] (ii) a light chain variable (VL) region incorporating the following CDRs:

[1001] LC-CDR1 having the amino acid sequence of SEQ ID NO:179 LC-CDR2 having the amino acid sequence of SEQ ID NQ:180 LC-CDR3 having the amino acid sequence of SEQ ID NO:173; optionally wherein the antigen-binding moiety comprises a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:165; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:178; further optionally wherein the antigen-binding moiety is an scFv; and wherein SNAP is a SNAP protein, optionally comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:419.

[1002] In some embodiments, the antigen-binding molecule is scFV(C008)-SNAP-Auristatin F and the DAR is 1.

[1003] In some embodiments, the scFv(C008)-SNAP component of the antigen-binding molecule comprises an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:417.

[1004] The present disclosure also provides compositions comprising an antigen-binding molecule according to the present disclosure.

[1005] The antigen-binding molecules described herein may be formulated as pharmaceutical compositions or medicaments for clinical use and may comprise a pharmaceutically acceptable carrier, diluent, excipient or adjuvant. Thus, the present disclosure provides a pharmaceutical composition / medicament comprising an antigen-binding molecule described herein.

[1006] The pharmaceutical compositions / medicaments of the present disclosure may comprise one or more pharmaceutically-acceptable carriers (e.g. liposomes, micelles, microspheres, nanoparticles), diluents / excipients (e.g. starch, cellulose, a cellulose derivative, a polyol, dextrose, maltodextrin, magnesium stearate), adjuvants, fillers, buffers, preservatives (e.g. vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methyl paraben, propyl paraben), anti-oxidants (e.g. vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium), lubricants (e.g. magnesium stearate, talc, silica, stearic acid, vegetable stearin), binders (e.g. sucrose, lactose, starch, cellulose, gelatin, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), xylitol, sorbitol, mannitol), stabilisers, solubilisers, surfactants (e.g., wetting agents), masking agents or colouring agents (e.g. titanium oxide).

[1007] The term ‘pharmaceutically-acceptable’ as used herein pertains to compounds, ingredients, materials, compositions, dosage forms, etc., which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of the subject in question (e.g. a human subject) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, diluent, excipient, adjuvant, filler, buffer, preservative, anti-oxidant, lubricant, binder, stabiliser, solubiliser, surfactant, masking agent, colouring agent, flavouring agent or sweetening agent of a composition according to the present disclosure must also be ‘acceptable’ in the sense of being compatible with the other ingredients of the formulation. Suitable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, anti-oxidants, lubricants, binders, stabilisers, solubilisers, surfactants, masking agents, colouring agents, flavouring agents or sweetening agents can be found in standard pharmaceutical texts, for example, Remington’s ‘The Science and Practice of Pharmacy’ (Ed. A. Adejare), 23rdEdition (2020), Academic Press.

[1008] Pharmaceutical compositions and medicaments of the present disclosure may be formulated for topical, parenteral, systemic, intracavitary, intravenous, intra-arterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, intrathecal, oral or transdermal routes of administration. In some embodiments, a pharmaceutical composition / medicament may be formulated for administration by injection or infusion, or administration by ingestion.

[1009] Suitable formulations may comprise the antigen-binding molecule provided in a sterile or isotonic medium. Medicaments and pharmaceutical compositions may be formulated in fluid, including gel, form. Fluid formulations may be formulated for administration by injection or infusion (e.g. via catheter) to a selected region of the human or animal body.

[1010] In some embodiments, the pharmaceutical compositions / medicament is formulated for injection or infusion, e.g. into a blood vessel, tissue / organ of interest, or tumor.

[1011] The present disclosure also provides methods for the production of pharmaceutically useful compositions, such methods of production may comprise one or more steps selected from: producing an antigenbinding molecule described herein; isolating / purifying an antigen-binding molecule described herein; and / or mixing an antigen-binding molecule described herein with a pharmaceutically-acceptable carrier, adjuvant, excipient or diluent.

[1012] For example, a further aspect the present disclosure relates to a method of formulating or producing a medicament or pharmaceutical composition for use in the treatment of a disease / condition (e.g. a disease / condition described herein), the method comprising formulating a pharmaceutical composition or medicament by mixing an antigen-binding molecule described herein with a pharmaceutically acceptable carrier, adjuvant, excipient or diluent.

[1013] Therapeutic and prophylactic applications

[1014] The antigen-binding molecules and compositions described herein find use in therapeutic and prophylactic methods.

[1015] The present disclosure provides an antigen-binding molecule or composition described herein for use in a method of medical treatment or prophylaxis. The present disclosure provides an antigen-binding molecule or composition described herein for use as a medicament. For example, the present disclosure provides an antigen-binding molecule that binds to CNX, comprising (i) a CNX-binding moiety and (ii) a linkerpayload moiety, for use as a medicament.

[1016] Also provided is an antigen-binding molecule or composition described herein for use in a method of treating or preventing a disease or condition described herein. Also provided is the use of an antigenbinding molecule or composition described herein in the manufacture of a medicament for treating or preventing a disease or condition described herein. Also provided is a method of treating or preventing a disease or condition described herein, comprising administering to a subject a therapeutically or prophylactically effective amount of an antigen-binding molecule or composition described herein.

[1017] The methods may be effective to reduce the development or progression of a disease / condition, alleviation of the symptoms of a disease / condition or reduction in the pathology of a disease / condition. The methods may be effective to prevent progression of the disease / condition, e.g. to prevent worsening of, or to slow the rate of development of, the disease / condition. In some embodiments, the methods may lead to an improvement in the disease / condition, e.g. a reduction in the symptoms of the disease / condition or reduction in some other correlate of the severity / activity of the disease / condition. In some embodiments, the methods may prevent development of the disease / condition a later stage (e.g. a chronic stage or metastasis).

[1018] It will be appreciated that the antigen-binding molecules or compositions of the present disclosure may be used for the treatment / prevention of any disease / condition that would derive therapeutic or prophylactic benefit from a reduction in the level / activity of CNX, CRT, complexes comprising CNX / CRT, or a reduction in the number or activity of cells comprising / expressing CNX, CRT, or complexes comprising CNX / CRT.

[1019] For example, the disease / condition may be a disease / condition in which CNX, CRT, complexes comprising CNX / CRT, or cells expressing / expressing the same are pathologically-implicated, e.g. a disease / condition in which an increased level / activity of CNX, CRT, complexes comprising CNX / CRT, of an increase in the number / proportion of cells comprising / expressing CNX, CRT or complexes comprising CNX / CRT is positively associated with the onset, development or progression of the disease / condition, and / or severity of one or more symptoms of the disease / condition. In some embodiments, an increased level / activity of CNX, CRT, complexes comprising CNX / CRT, of an increase in the number / proportion of cells comprising / expressing CNX, CRT or complexes comprising CNX / CRT may be a risk factor for the onset, development or progression of the disease / condition.

[1020] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterised by an increase in the level of expression or activity of CNX, CRT or complexes comprising CNX or CRT, e.g. as compared to the level of expression / activity in the absence of the disease / condition. In some embodiments, the disease / condition to be treated / prevented is a disease / condition characterised by an increase in the number / proportion / activity of cells expressing CNX, CRT or complexes comprising CNX or CRT, e.g. as compared to the level / number / proportion / activity in the absence of the disease / condition.

[1021] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterised by an increase in the level of surface expression of CNX, CRT or complexes comprising CNX or CRT, e.g. as compared to the level of surface expression in the absence of the disease / condition.

[1022] For example, where the disease / condition is a cancer, the cancer may comprise cells having increased surface expression of CNX, CRT or complexes comprising CNX or CRT. Where the disease / condition is a cancer, the level of surface expression of CNX, CRT or complexes comprising CNX or CRT on a cell may be greater than the level of surface expression of CNX, CRT or complexes comprising CNX or CRT on equivalent non-cancerous cells / non-tumor tissue.

[1023] In some embodiments, the disease / condition to be treated / prevented is a disease / condition characterised by an increase in the number / proportion of cells expressing CNX, CRT or complexes comprising CNX or CRT on the cell surface, e.g. as compared to the number / proportion of cells in the absence of the disease / condition.

[1024] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterised by increased levels of GALA activation, e.g. as compared to the level of GALA activation in the absence of the disease / condition.

[1025] For example, where the disease / condition is a cancer, the cancer may comprise cells having increased levels of GALA activation. Where the disease / condition is a cancer, the level of GALA activation may be greater than the level of GALA activation in equivalent non-cancerous cells / non-tumor tissue.

[1026] In some embodiments, the disease / condition to be treated / prevented is a disease / condition characterised by an increase in the number / proportion of cells with increased levels of GALA activation e.g. as compared to the number / proportion of cells in the absence of the disease / condition.

[1027] The GalNAc-T activation (GALA) pathway is a cellular process in which GalNAc-T (GALNT) enzymes are relocated from the Golgi to the endoplasmic reticulum (ER). Trafficking of GALNTs to the ER may be stimulated by growth factors (e.g. EGF, PDGF) and activation of Src kinase. GALNTs are active in the ER and GalNAc incorporation in proteins increases after GALNT relocation. GALA activation may be evaluated, for example, by detecting the level of a correlate of GALA activation (e.g. O-glycosylation, Tn glycosylation, Src activity, GALNT localisation to the ER, GALNT activity). GALA activation may be evaluated in an appropriate assay. Such assays are known to the skilled person and include e.g. in vitro or ex vivo assays.

[1028] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition described in WO 2020 / 159445 A1 (hereby incorporated by reference in its entirety).

[1029] Treatment in accordance with the methods of the present disclosure may achieve one or more of the following in a subject (compared to an equivalent untreated subject, or subject treated with an appropriate control): a reduction in the level of CNX, CRT or complexes comprising CNX or CRT ; a reduction in the activity of CNX, CRT or complexes comprising CNX or CRT ; and / or a reduction in the number / proportion of cells comprising / expressing CNX, CRT or complexes comprising CNX or CRT.

[1030] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated O-glycosylation activity. For example, where the disease / condition is a cancer, the cancer may comprise cells having elevated O-glycosylation activity. As used herein ‘O- glycosylation activity’ refers to addition of O-linked glycan to the hydroxyl group of the side chain of e.g. a serine, threonine, tyrosine, hydroxylysine, or hydroxy proline residue of a protein. An ‘elevated’ level of O- glycosylation activity may refer to a level of O-glycosylation activity which is greater than the level of O- glycosylation activity in the absence of the disease / condition (e.g. in a healthy subject, or in equivalent non-diseased tissue). Where the disease / condition is a cancer, the level of O-glycosylation activity may be greater than the level of O-glycosylation activity in equivalent non-cancerous cells / non-tu mor tissue. A cancer / cell thereof may comprise one or more mutations (e.g. relative to equivalent non-cancerous cells / non-tumor tissue) causing upregulation of O-glycosylation activity.

[1031] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated Src activity. For example, where the disease / condition is a cancer, the cancer may comprise cells having elevated Src activity. As used herein ‘Src activity’ refers to Src- mediated phosphorylation of tyrosine residues. An ‘elevated’ level of Src activity may refer to a level of Src activity which is greater than the level of Src activity in the absence of the disease / condition (e.g. in a healthy subject, or in equivalent non-diseased tissue). Where the disease / condition is a cancer, the level of Src activity may be greater than the level of Src activity in equivalent non-cancerous cells / non-tumor tissue. A cancer / cell thereof may comprise one or more mutations (e.g. relative to equivalent non- cancerous cells / non-tumor tissue) causing upregulation of Src activity.

[1032] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated GalNAc-transferase (GALNT) activity. For example, where the disease / condition is a cancer, the cancer may comprise cells having elevated GALNT activity. As used herein ‘GALNT activity’ refers to GALNT-mediated transfer of N-acetylgalactosamine (GalNAc) from LIDP- GalNAc to the hydroxyl group of the side chain of e.g. a serine or threonine residue. An ‘elevated’ level of GALNT activity may refer to a level of GALNT activity which is greater than the level of GALNT activity in the absence of the disease / condition (e.g. in a healthy subject, or in equivalent non-diseased tissue). Where the disease / condition is a cancer, the level of GALNT activity may be greater than the level of GALNT activity in equivalent non-cancerous cells / non-tumor tissue. A cancer / cell thereof may comprise one or more mutations (e.g. relative to equivalent non-cancerous cells / non-tumor tissue) causing upregulation of GALNT activity.

[1033] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated O-glycosylation. For example, where the disease / condition is a cancer, the cancer may comprise cells having an elevated level of O-glycosylation of a protein expressed by the cells. An ‘elevated’ level of O-glycosylation may refer to a level of O-glycosylation which is greater than the level of O-glycosylation in the absence of the disease / condition (e.g. in a healthy subject, or in equivalent non-diseased tissue). Where the disease / condition is a cancer, the level of O-glycosylation may be greater than the level of O-glycosylation in equivalent non-cancerous cells / non-tumor tissue. A cancer / cell thereof may comprise one or more mutations (e.g. relative to equivalent non-cancerous cells / non-tumor tissue) causing upregulation of O-glycosylation.

[1034] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated Tn glycosylation. For example, where the disease / condition is a cancer, the cancer may comprise cells having Tn glycosylation of a protein expressed by the cells. As used herein ‘Tn glycosylation’ refers to the presence of N-acetylgalactosamine (GalNAc) linked to the hydroxyl group of the side chain of a serine or threonine residue of a protein by a glycosidic bond. A ‘Tn glycosylated’ protein comprises at least one Tn glycan, which may also be referred to as Tn antigen.

[1035] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated Tn glycosylation. For example, where the disease / condition is a cancer, the cancer may comprise cells having an elevated level of Tn glycosylation of a protein expressed by the cells. An ‘elevated’ level of Tn glycosylation may refer to a level of Tn glycosylation which is greater than the level of Tn glycosylation in the absence of the disease / condition (e.g. in a healthy subject, or in equivalent non-diseased tissue). Where the disease / condition is a cancer, the level of Tn glycosylation may be greater than the level of Tn glycosylation in equivalent non-cancerous cells / non- tumor tissue. A cancer / cell thereof may comprise one or more mutations (e.g. relative to equivalent non- cancerous cells / non-tumor tissue) causing upregulation of Tn glycosylation. A protein having an ‘elevated’ level of Tn glycosylation as compared to a reference protein may possess more Tn glycans than the reference protein.

[1036] The anti-CNX antibodies of the present disclosure are demonstrated to be useful to inhibit ECM degradation. Accordingly, in some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterised by extracellular matrix (ECM) degradation. A disease / condition which is ‘characterised by ECM degradation’ may be a disease / condition in which ECM degradation is a symptom of the disease / condition. The disease / condition to be treated / prevented in accordance with the present disclosure may be a disease / condition in which ECM degradation is pathologically-implicated. For example, the disease / condition may be a disease / condition in which ECM degradation, and / or an increased level of ECM degradation, is implicated in the pathology of the disease / condition.

[1037] Diseases / conditions characterised by ECM degradation include e.g. cancers.

[1038] The involvement of ECM degradation in the development and progression of cancers is well known, and is reviewed e.g. in Walker et al., Int. J. Mol. Sci. (2018) 19(10): 3028, Najafi et al., J. Cell Biochem. (2019) 120(3):2782-2790 and Winkler et al., Nat. Commun. (2020) 11 (1 ): 5120, all of which are hereby incorporated by reference in their entirety.

[1039] In some embodiments the disease / condition to be treated / prevented is a cancer. Cancer may refer to any unwanted cell proliferation (or any disease manifesting itself by unwanted cell proliferation), neoplasm or tumor. The cancer may be benign or malignant and may be primary or secondary (metastatic). A neoplasm or tumor may be any abnormal growth or proliferation of cells and may be located in any tissue. The cancer may be of tissues / cells derived from e.g. the adrenal gland, adrenal medulla, anus, appendix, bladder, blood, bone, bone marrow, brain, breast, cecum, central nervous system (including or excluding the brain) cerebellum, cervix, colon, duodenum, endometrium, epithelial cells (e.g. renal epithelia), gallbladder, oesophagus, glial cells, heart, ileum, jejunum, kidney, lacrimal glad, larynx, liver, lung, lymph, lymph node, lymphoblast, maxilla, mediastinum, mesentery, myometrium, nasopharynx, omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous system, peritoneum, pleura, prostate, salivary gland, sigmoid colon, skin, small intestine, soft tissues, spleen, stomach, testis, thymus, thyroid gland, tongue, tonsil, trachea, uterus, vulva, and / or white blood cells.

[1040] Tumors may be nervous or non-nervous system tumors. Nervous system tumors may originate either in the central or peripheral nervous system, e.g. glioma, medulloblastoma, meningioma, neurofibroma, ependymoma, Schwannoma, neurofibrosarcoma, astrocytoma and oligodendroglioma. Non-nervous system cancers / tumors may originate in any other non-nervous tissue, examples include melanoma, mesothelioma, lymphoma, myeloma, leukemia, Non-Hodgkin’s lymphoma (NHL), Hodgkin’s lymphoma, chronic myelogenous leukemia (CML), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), cutaneous T cell lymphoma (CTCL), chronic lymphocytic leukemia (CLL), hepatoma, epidermoid carcinoma, prostate carcinoma, breast cancer, lung cancer, colon cancer, ovarian cancer, pancreatic cancer, thymic carcinoma, NSCLC, hematologic cancer and sarcoma.

[1041] Anti-CNX antibodies are shown herein and also e.g. in Ros et al. Nat. Cell Biol. (2020) 22(11 ): 1371 -1381 and WO 2020 / 159445 A1 to be useful to inhibit tumor growth and metastasis, including of breast and liver cancers. Ryan et al., J. Transl. Med. (2016) 14:196 proposes CNX as a therapeutic target in colorectal cancer. ADCs comprising an anti-CNX antigen-binding moiety are shown herein to be useful to inhibit tumour growth and metastasis, including of liver, pancreatic and breast cancers. Chen et al., Cancer Immunol. Res. (2019) 7(1 ):123-135 demonstrate that expression of CNX is upregulated in oral squamous cell carcinoma, and that knockdown of CNX improved control of tumor growth in a model of melanoma. The authors also demonstrated that CNX inhibits the proliferation and effector function of CD4+ and CD8+ T cells, by a mechanism involving upregulation of the expression of immune checkpoint molecule PD-1 . Thus, Chen et al. suggests that intervention targeting CNX could be useful for the treatment / prevention of a wide range of cancers, through indirect antagonism of PD-1 / PD- L1-mediated suppression of anticancer responses.

[1042] In some embodiments, the cancer is liver cancer, breast cancer, oral cancer (e.g. oral squamous cell carcinoma), sarcoma, lung cancer, prostate cancer, bladder cancer, renal cancer, melanoma, pancreatic cancer, endometrial cancer, colorectal cancer, thyroid cancer, cervical cancer, or gastric cancer. In some embodiments, the cancer is cervical cancer, liver cancer (e.g. hepatocellular carcinoma, bile duct cancer), breast cancer (e.g. triple negative breast cancer), pancreatic cancer, gastric cancer, or colorectal cancer.

[1043] In some embodiments, the liver cancer is a primary liver cancer. In some embodiments, the liver cancer is hepatocellular carcinoma (HCC), fibrolamellar carcinoma, bile duct cancer (cholangiocarcinoma), angiosarcoma or hepatoblastoma.

[1044] In some embodiments, the breast cancer is a primary breast cancer. In some embodiments, the breast cancer is ductal carcinoma, lobular carcinoma, in situ breast cancer (e.g. ductal carcinoma in situ (DCIS) invasive carcinoma (e.g. invasive ductal carcinoma (IDC), invasive lobular carcinoma (ILC), triple negative breast cancer or inflammatory breast cancer), Paget disease, angiosarcoma or Phyllodes tumor.

[1045] In some embodiments, the cancer is a cancer that would derive therapeutic or prophylactic benefit from a reduction in the expression or activity of CNX, CRT or complexes comprising CNX or CRT. In some embodiments, the cancer is a cancer which is caused or exacerbated by expression / overexpression or activity of CNX, CRT or complexes comprising CNX or CRT. In some embodiments, the cancer is a cancer for which expression / overexpression or activity of CNX, CRT or complexes comprising CNX or CRT is a risk factor for the development or progression of the cancer. In some embodiments the cancer is a cancer for which expression / overexpression or activity of CNX, CRT or complexes comprising CNX or CRT is positively associated with onset, development, progression, severity or metastasis.

[1046] As used herein, overexpression of a given protein / protein complex (e.g. CNX, CRT, complexes comprising the same) refers to a level of gene or protein expression of the relevant protein / protein complex which is greater than the level of expression by equivalent non-cancerous cells / non-tumor tissue.

[1047] In some embodiments, the cancer may be a cancer characterised by expression / overexpression of CNX or CRT (i.e. ‘CNX-positive’ cancers and ‘CRT-positive’ cancers, respectively), or of polypeptide complexes comprising CNX / CRT. The cancer may comprise cells expressing / overexpressing CNX, CRT, or polypeptide complexes comprising CNX / CRT. CNX / CRT expression may be determined by any suitable means. Expression may be gene expression or protein expression. Gene expression can be determined e.g. by detection of mRNA encoding CNX / CRT, for example by quantitative real-time PCR (qRT-PCR). Protein expression can be determined e.g. by detection of CNX / CRT, for example by antibody-based methods, for example by western blot, immunohistochemistry, immunocytochemistry, flow cytometry, or ELISA.

[1048] In some embodiments, the cancer may be a cancer characterised by surface expression of CNX / CRT. In some embodiments, the cancer may comprise cells expressing CNX / CRT at the cell surface. CNX / CRT may be present in or at the cell membrane of cells of the cancer.

[1049] In some embodiments, the cancer may be a cancer characterised by expression / overexpression of O- glycosylated CNX / CRT. The cancer may comprise cells expressing / overexpressing O-glycosylated CNX / CRT. In some embodiments, the cancer may be a cancer characterised by expression of CNX / CRT having an elevated level of O-glycosylation. The cancer may comprise cells expressing CNX / CRT having an elevated level of O-glycosylation.

[1050] In some embodiments, the cancer may be a cancer characterised by expression / overexpression of Tn glycosylated CNX / CRT. The cancer may comprise cells expressing / overexpressing Tn glycosylated CNX / CRT. In some embodiments, the cancer may be a cancer characterised by expression of CNX / CRT having an elevated level of Tn glycosylation. The cancer may comprise cells expressing CNX / CRT having an elevated level of Tn glycosylation.

[1051] Treatment of a subject with an antigen-binding molecule in accordance with the present disclosure may: delay / prevent the onset of one or more symptoms of the cancer; reduce the severity of one or more symptoms of the cancer; increase survival of the subject; reduce / inhibit survival of cells of the cancer; reduce the number of cells of the cancer in the subject; reduce tumor size / volume; reduce cancer / tumor burden in the subject; reduce / inhibit growth of cells of the cancer; reduce / inhibit tumor growth; reduce / inhibit invasion by cells of the cancer; and / or reduce / inhibit metastasis of the cancer.

[1052] Administration of the antigen-binding molecules and compositions of the present disclosure is preferably in a ‘therapeutically-effective’ or ‘prophylactically-effective’ amount, this being sufficient to show therapeutic or prophylactic benefit to the subject. The actual amount administered, and rate and timecourse of administration, will depend on the nature and severity of the disease / condition and the particular article administered. Prescription of treatment, e.g. decisions on dosage etc., is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disease / disorder to be treated, the condition of the individual subject, the site of delivery, the method of administration and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington’s ‘The Science and Practice of Pharmacy’ (ed. A. Adejare), 23rdEdition (2020), Academic Press. Administration of the antigen-binding molecules and compositions of the present disclosure may be topical, parenteral, systemic, intracavitary, intravenous, intra-arterial, intramuscular, intrathecal, intraocular, intravitreal, intraconjunctival, subretinal, suprachoroidal, subcutaneous, intradermal, intrathecal, oral, nasal or transdermal. Administration may be by injection or infusion. Administration may be intratumoral.

[1053] In some aspects and embodiments, articles of the present disclosure may be administered to a tissue / organ of interest (e.g. a tissue / organ affected by the disease / condition, e.g. a tissue / organ in which symptoms of the disease / condition manifest). In some aspects and embodiments, articles of the present disclosure may be administered to the blood ( / .e. intravenous / intra-arterial administration) by injection or infusion (e.g. via cannula), or may be administered subcutaneously or orally. In some aspects and embodiments, articles of the present disclosure may be administered to a tumor.

[1054] Administration may be alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated. The antigen-binding molecule or composition described herein and a therapeutic agent may be administered simultaneously or sequentially.

[1055] In some embodiments, the methods comprise additional therapeutic or prophylactic intervention, e.g. for the treatment / prevention of a cancer. In some embodiments, the therapeutic or prophylactic intervention is selected from chemotherapy, immunotherapy, radiotherapy, surgery, vaccination and / or hormone therapy. In some embodiments, the therapeutic or prophylactic intervention comprises leukapheresis. In some embodiments, the therapeutic or prophylactic intervention comprises a stem cell transplant.

[1056] Simultaneous administration refers to administration of the antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell or composition and therapeutic agent together, for example as a pharmaceutical composition containing both agents (combined preparation), or immediately after each other and optionally via the same route of administration, e.g. to the same artery, vein or other blood vessel. Sequential administration refers to administration of one of the antigen-binding molecule / composition or therapeutic agent followed after a given time interval by separate administration of the other agent. It is not required that the two agents are administered by the same route, although this is the case in some embodiments. The time interval may be any time interval.

[1057] Chemotherapy and radiotherapy respectively refer to treatment of a cancer with a drug or with ionising radiation (e.g. radiotherapy using X-rays or y-rays). The drug may be a chemical entity, e.g. small molecule pharmaceutical, antibiotic, DNA intercalator, protein inhibitor (e.g. kinase inhibitor), or a biological agent, e.g. antibody, antibody fragment, aptamer, nucleic acid (e.g. DNA, RNA), peptide, polypeptide, or protein.

[1058] The chemotherapy may be administered according to a treatment regime. The treatment regime may be a pre-determined timetable, plan, scheme or schedule of chemotherapy administration which may be prepared by a physician or medical practitioner and may be tailored to suit the patient requiring treatment. The treatment regime may indicate one or more of: the type of chemotherapy to administer to the patient; the dose of each drug or radiation; the time interval between administrations; the length of each treatment; the number and nature of any treatment holidays, if any etc. For a co-therapy a single treatment regime may be provided which indicates how each drug is to be administered.

[1059] Chemotherapeutic drugs may be selected from: Abemaciclib, Abiraterone Acetate, Abitrexate (Methotrexate), Abraxane (Paclitaxel Albumin-stabilized Nanoparticle Formulation), ABVD, ABVE, ABVE- PC, AC, Acalabrutinib, AC-T, Adcetris (Brentuximab Vedotin), ADE, Ado-Trastuzumab Emtansine, Adriamycin (Doxorubicin Hydrochloride), Afatinib Dimaleate, Afinitor (Everolimus), Akynzeo (Netupitant and Palonosetron Hydrochloride), Aldara (Imiquimod), Aldesleukin, Alecensa (Alectinib), Alectinib, Alemtuzumab, Alimta (Pemetrexed Disodium), Aliqopa (Copanlisib Hydrochloride), Alkeran for Injection (Melphalan Hydrochloride), Alkeran Tablets (Melphalan), Aloxi (Palonosetron Hydrochloride), Alunbrig (Brigatinib), Ambochlorin (Chlorambucil), Amboclorin (Chlorambucil), Amifostine, Aminolevulinic Acid, Anastrozole, Aprepitant, Aredia (Pamidronate Disodium), Arimidex (Anastrozole), Aromasin (Exemestane), Arranon (Nelarabine), Arsenic Trioxide, Arzerra (Ofatumumab), Asparaginase Erwinia chrysanthemi, Atezolizumab, Avastin (Bevacizumab), Avelumab, Axicabtagene Ciloleucel, Axitinib, Azacitidine, Bavencio (Avelumab), BEACOPP, Becenum (Carmustine), Beleodaq (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, Besponsa (Inotuzumab Ozogamicin) , Bevacizumab, Bexarotene, Bexxar (Tositumomab and Iodine I 131 Tositumomab), Bicalutamide, BiCNU (Carmustine), Bleomycin, Blinatumomab, Blincyto (Blinatumomab), Bortezomib, Bosulif (Bosutinib), Bosutinib, Brentuximab Vedotin, Brigatinib, BuMel, Busulfan, Busulfex (Busulfan), Cabazitaxel, Cabometyx (Cabozantinib-S- Malate), Cabozantinib-S-Malate, CAF, Calquence (Acalabrutinib), Campath (Alemtuzumab), Camptosar (Irinotecan Hydrochloride), Capecitabine, CAPOX, Carac (Fluorouracil — Topical), Carboplatin, CARBOPLATIN-TAXOL, Carfilzomib, Carmubris (Carmustine), Carmustine, Carmustine Implant, Casodex (Bicalutamide), CEM, Ceritinib, Cerubidine (Daunorubicin Hydrochloride), Cervarix (Recombinant HPV Bivalent Vaccine), Cetuximab, CEV, Chlorambucil, CHLORAMBUCIL-PREDNISONE, CHOP, Cisplatin, Cladribine, Clafen (Cyclophosphamide), Clofarabine, Clofarex (Clofarabine), Clolar (Clofarabine), CMF, Cobimetinib, Cometriq (Cabozantinib-S-Malate), Copanlisib Hydrochloride, COPDAC, COPP, COPP-ABV, Cosmegen (Dactinomycin), Cotellic (Cobimetinib), Crizotinib, CVP, Cyclophosphamide, Cyfos (Ifosfamide), Cyramza (Ramucirumab), Cytarabine, Cytarabine Liposome, Cytosar-U (Cytarabine), Cytoxan (Cyclophosphamide), Dabrafenib, Dacarbazine, Dacogen (Decitabine), Dactinomycin, Daratumumab, Darzalex (Daratumumab), Dasatinib, Daunorubicin Hydrochloride, Daunorubicin Hydrochloride and Cytarabine Liposome, Decitabine, Defibrotide Sodium, Defitelio (Defibrotide Sodium), Degarelix, Denileukin Diftitox, Denosumab, DepoCyt (Cytarabine Liposome), Dexamethasone, Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, Doxil (Doxorubicin Hydrochloride Liposome), Doxorubicin Hydrochloride, Doxorubicin Hydrochloride Liposome, Dox-SL (Doxorubicin Hydrochloride Liposome), DTIC-Dome (Dacarbazine), Durvalumab, Efudex (Fluorouracil — Topical), Elitek (Rasburicase), Ellence (Epirubicin Hydrochloride), Elotuzumab, Eloxatin (Oxaliplatin), Eltrombopag Olamine, Emend (Aprepitant), Empliciti (Elotuzumab), Enasidenib Mesylate, Enzalutamide, Epirubicin Hydrochloride, EPOCH, Erbitux (Cetuximab), Eribulin Mesylate, Erivedge (Vismodegib), Erlotinib Hydrochloride, Erwinaze (Asparaginase Erwinia chrysanthemi), Ethyol (Amifostine), Etopophos (Etoposide Phosphate), Etoposide, Etoposide Phosphate, Evacet (Doxorubicin Hydrochloride Liposome), Everolimus, Evista (Raloxifene Hydrochloride), Evomela (Melphalan Hydrochloride), Exemestane, 5-FU (Fluorouracil Injection), 5-FU (Fluorouracil — Topical), Fareston (Toremifene), Farydak (Panobinostat), Faslodex (Fulvestrant), FEC, Femara (Letrozole), Filgrastim, Fludara (Fludarabine Phosphate), Fludarabine Phosphate, Fluoroplex (Fluorouracil — Topical), Fluorouracil Injection, Fluorouracil — Topical, Flutamide, Folex (Methotrexate), Folex PFS (Methotrexate), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, Folotyn (Pralatrexate), FU-LV, Fulv...

Claims

Claims:

1. An antigen-binding molecule that binds to CNX, comprising (i) a CNX-binding moiety and (ii) a linkerpayload moiety.

2. The antigen-binding molecule according to claim 1 , wherein the CNX-binding moiety comprises:(a)(i) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and(II) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; or(b)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:179 LC-CDR2 having the amino acid sequence of SEQ ID NQ:180 LC-CDR3 having the amino acid sequence of SEQ ID NO:173; or(c)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:4; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NQ:10 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NO:12; or(d)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:18 HC-CDR2 having the amino acid sequence of SEQ ID NO:19 HC-CDR3 having the amino acid sequence of SEQ ID NQ:20; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:25 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:27; or(e)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:49; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:53 LC-CDR2 having the amino acid sequence of SEQ ID NO:54 LC-CDR3 having the amino acid sequence of SEQ ID NO:55; or(f)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:62 HC-CDR3 having the amino acid sequence of SEQ ID NO:63; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:68 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:69; or(g)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:62 HC-CDR3 having the amino acid sequence of SEQ ID NO:63; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:73 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:74; or(h)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:62 HC-CDR3 having the amino acid sequence of SEQ ID NO:63; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:78 LC-CDR2 having the amino acid sequence of SEQ ID NO:79 LC-CDR3 having the amino acid sequence of SEQ ID NQ:80; or(i)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:83; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:73LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:74; or(j)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:86; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:89 LC-CDR2 having the amino acid sequence of SEQ ID NO:11 LC-CDR3 having the amino acid sequence of SEQ ID NQ:90; or(k)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:61 HC-CDR2 having the amino acid sequence of SEQ ID NO:95 HC-CDR3 having the amino acid sequence of SEQ ID NO:96; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NQ:101 LC-CDR2 having the amino acid sequence of SEQ ID NQ:102 LC-CDR3 having the amino acid sequence of SEQ ID NQ:103; or(l)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NQ:108 HC-CDR2 having the amino acid sequence of SEQ ID NQ:109 HC-CDR3 having the amino acid sequence of SEQ ID NQ:110; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:115 LC-CDR2 having the amino acid sequence of SEQ ID NO:116 LC-CDR3 having the amino acid sequence of SEQ ID NO:117; or(m)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:122; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:125 LC-CDR2 having the amino acid sequence of SEQ ID NO:126 LC-CDR3 having the amino acid sequence of SEQ ID NO:127; or(n)(I) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO:132 HC-CDR2 having the amino acid sequence of SEQ ID NO:133HC-CDR3 having the amino acid sequence of SEQ ID NO:134; and(ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:139 LC-CDR2 having the amino acid sequence of SEQ ID NQ:140 LC-CDR3 having the amino acid sequence of SEQ ID NQ:80; or(o)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:146 HC-CDR2 having the amino acid sequence of SEQ ID NO:147 HC-CDR3 having the amino acid sequence of SEQ ID NO:148; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:153; or (P)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:3 HC-CDR3 having the amino acid sequence of SEQ ID NO:156; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:158 LC-CDR2 having the amino acid sequence of SEQ ID NO:159 LC-CDR3 having the amino acid sequence of SEQ ID NQ:160; or(q)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:171 LC-CDR2 having the amino acid sequence of SEQ ID NO:172 LC-CDR3 having the amino acid sequence of SEQ ID NO:173; or(r)(I) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:185 HC-CDR2 having the amino acid sequence of SEQ ID NO:186 HC-CDR3 having the amino acid sequence of SEQ ID NO:187; and(II) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:73 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:194; or(s)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:199 HC-CDR3 having the amino acid sequence of SEQ ID NQ:200; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NQ:205 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NQ:206; or(t)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:211 HC-CDR3 having the amino acid sequence of SEQ ID NO:212; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:216 LC-CDR2 having the amino acid sequence of SEQ ID NO:172 LC-CDR3 having the amino acid sequence of SEQ ID NO:217; or(u)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:222 HC-CDR2 having the amino acid sequence of SEQ ID NO:223 HC-CDR3 having the amino acid sequence of SEQ ID NO:224; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:229 LC-CDR2 having the amino acid sequence of SEQ ID NO:172 LC-CDR3 having the amino acid sequence of SEQ ID NQ:230; or(v)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:48 HC-CDR2 having the amino acid sequence of SEQ ID NO:199 HC-CDR3 having the amino acid sequence of SEQ ID NQ:200; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:235 LC-CDR2 having the amino acid sequence of SEQ ID NO:236 LC-CDR3 having the amino acid sequence of SEQ ID NO:237; or(w)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:185 HC-CDR2 having the amino acid sequence of SEQ ID NO:243 HC-CDR3 having the amino acid sequence of SEQ ID NO:244; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:248LC-CDR2 having the amino acid sequence of SEQ ID NO:249LC-CDR3 having the amino acid sequence of SEQ ID NO:250.

3. The antigen-binding molecule according to claim 1 or claim 2, wherein the CNX-binding moiety comprises: a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32, 165, 1 , 17, 47, 60, 82, 85, 94, 107, 121 , 131 , 154, 155, 184, 198, 210, 221 or 242; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:40, 178, 9, 24, 52, 67, 72, 77, 88, 100, 114, 124, 138, 152, 157, 170, 191, 204, 215, 228, 234 or 247; optionally4. The antigen-binding molecule according to any one of claims 1 to 3, wherein the CNX-binding moiety comprises:(i) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:32; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:40; or(II) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:165; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:178; or(ill) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:1; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:9; or(iv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:17; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:24; or(v) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:47; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:52; or(vi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:60; anda VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:67; or(vii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:60; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:72; or(viii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:60; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:77; or(ix) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:82; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:72; or(x) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:85; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:88; or(xi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:94; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:100; or(xii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:107; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:114; or(xiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:121 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:124; or(xiv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:131 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:138;or(xv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:145; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:152; or(xvi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:155; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:157; or(xvii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:165; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:170; or(xviii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:184; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:191; or(xix) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:198; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:204; or(xx) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:210; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:215; or(xxi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:221 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:228; or(xxii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:198; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:234; or(xxiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:242; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:247.

5. The antigen-binding molecule according to any one of claims 1 to 4, wherein the linker-payload moiety comprises or consists of a cytotoxic agent.

6. The antigen-binding molecule according to any one of claims 1 to 5, wherein the linker-payload moiety comprises or consists of: a microtubule-targeting agent, a DNA-targeting agent, an RNA-targeting agent, an immune system-activating agent, an apoptosis-promoting agent, a metabolism-inhibiting agent and a proteasome inhibiting agent.

7. The antigen-binding molecule according to any one of claims 1 to 6, wherein the linker-payload moiety comprises or consists of a microtubule-targeting agent.

8. The antigen-binding molecule according to claim 7, wherein the linker-payload moiety comprises or consists of an auristatin.

9. The antigen-binding molecule according to claim 8, wherein the linker-payload moiety comprises or consists of monomethyl auristatin E (MMAE) or auristatin F.

10. The antigen-binding molecule according to any one of claims 1 to 6, wherein the linker-payload moiety comprises or consists of a DNA-targeting agent.

11. The antigen-binding molecule according to claim 10, wherein the linker-payload moiety comprises or consists of a topoisomerase inhibitor, optionally wherein the topoisomerase inhibitor is a TOP1 inhibitor.

12. The antigen-binding molecule according to claim 11, wherein the linker-payload moiety comprises or consists of camptothecin or a derivative thereof.

13. The antigen-binding molecule according to claim 12, wherein the linker-payload moiety comprises or consists of deruxtecan (Dxd).

14. The antigen-binding molecule according to any one of claims 1 to 13, wherein the linker-payload moiety comprises cathepsin-cleavable linker.

15. The antigen-binding molecule according to claim 14, wherein the linker-payload moiety comprises a valine-citruline linker or a glycine-glycine-phenylalanine-glycine linker.

16. The antigen-binding molecule according to any one of claims 1 to 15, wherein the antigen-binding molecule is selected from one of:(a)wherein ABM is an antigen-binding moiety which comprises:(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:41LC-CDR2 having the amino acid sequence of SEQ ID NO:42LC-CDR3 having the amino acid sequence of SEQ ID NO:43;wherein ABM is an antigen-binding moiety which comprises:(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:33 HC-CDR2 having the amino acid sequence of SEQ ID NO:34 HC-CDR3 having the amino acid sequence of SEQ ID NO:35; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:41 LC-CDR2 having the amino acid sequence of SEQ ID NO:42 LC-CDR3 having the amino acid sequence of SEQ ID NO:43; or(c)wherein SNAP is a SNAP protein, and wherein ABM is an antigen-binding moiety which comprises:(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:166 HC-CDR2 having the amino acid sequence of SEQ ID NO:167 HC-CDR3 having the amino acid sequence of SEQ ID NO:168; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:179 LC-CDR2 having the amino acid sequence of SEQ ID NQ:180 LC-CDR3 having the amino acid sequence of SEQ ID NO:173.

17. The antigen-binding molecule according to any one of claims 1 to 16, wherein the drug to antibody ratio (DAR) is between 1 and 8.

18. A composition comprising an antigen-binding molecule according to any one of claims 1 to 17, and a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

19. An antigen-binding molecule according to any one of claims 1 to 17, or a composition according to claim 18, for use as a medicament.

20. An antigen-binding molecule according to any one of claims 1 to 17, or a composition according to claim 18, for use in a method of medical treatment or prophylaxis, wherein the disease is characterised by cells which exhibit increased levels of GALA activation and / or increased cell surface expression of CNX.

21. An antigen-binding molecule according to any one of claims 1 to 17, or a composition according to claim 18, for use in treating or preventing a cancer.

22. Use of an antigen-binding molecule according to any one of claims 1 to 17, or a composition according to claim 18, in the manufacture of a medicament for treating or preventing a cancer.

23. A method of treating or preventing a cancer, comprising administering to a subject a therapeutically- or prophylactically-effective amount of an antigen-binding molecule according to any one of claims 1 to 17, or a composition according to claim 18.

24. The antigen-binding molecule or composition for use according to claim 21 , the use according to claim 22, or the method according to claim 23, wherein the cancer is selected from: liver cancer, breastcancer, oral cancer, oral squamous cell carcinoma, sarcoma, lung cancer, prostate cancer, bladder cancer, renal cancer, melanoma, pancreatic cancer, endometrial cancer, colorectal cancer, ovarian cancer, cervical cancer, brain cancer, bile duct cancer, testicular cancer, and thyroid cancer.

25. Use of antigen-binding molecule according to any one of claims 1 to 17 to deplete cells expressing CNX or increase killing of cells expressing CNX.

26. An in vitro complex, optionally isolated, comprising an antigen-binding molecule according to any one of claims 1 to 17 bound to CNX.

27. A method comprising contacting a cell expressing or suspected of expressing CNX with an antigenbinding molecule that binds to CNX, wherein the antigen binding molecule comprises (i) a CNX-binding moiety and (ii) a linker-payload moiety, wherein the antigen-binding molecule kills the cell.

28. A method of killing a cell expressing or suspected of expressing CNX, said method comprising contacting the cell with an antigen-binding molecule that binds to CNX, wherein the antigen binding molecule comprises (i) a CNX-binding moiety and (ii) a linker-payload moiety.

29. The method of claim 27 or claim 28, wherein the cell is a cancer cell.

30. The method of claim 29, wherein the cancer cell is a liver cancer cell, a pancreatic cancer cell or a breast cancer cell.

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