LRG1 antigen-binding molecules

Abstract

The present disclosure provides antigen-binding molecules capable of binding to LRG1. Also provided are nucleic acids encoding such antigen-binding molecules, vectors comprising such nucleic acids, cells comprising such antigen-binding molecules, and compositions comprising such antigen-binding molecules, nucleic acids, vectors, and / or cells. Further provided are methods for treating / preventing diseases / conditions using such antigen-binding molecules, nucleic acids, vectors, cells and / or compositions.

Description

[0001] LRG1 Antigen-Binding Molecules

[0002] This application claims priority from SG 10202402973X filed 25 September 2024, 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. The present disclosure also relates to methods of medical treatment and prophylaxis.

[0005] Background

[0006] LRG1 is a serum protein primarily produced by hepatocytes and neutrophils. Reportedly, LRG1 can also be secreted from endothelial cells, epithelial cells, fibroblasts, and other types of myeloid cells in local tissue, such as the lung, kidney, heart, skin, brain, and testis. The physiological role of LRG1 remains poorly understood due to the lack of overt phenotypic abnormalities in Lrg1 mice. However, LRG1 is thought to be involved in a plethora of physiological processes including the modulation of various signalling cascades, mainly TGFp signalling.

[0007] LRG1 is believed to play beneficial roles in the acute response to infection and injury. In particular, upregulated LRG1 expression in response to bacterial infection can mediate the differentiation and infiltration of lymphocytes and promote the survival of circulating immune cells by neutralizing cytochrome c cytotoxicity. In addition, LRG1 promotes wound healing or tissue repair by stimulating the renewal of damaged epithelial cells, tissue vascularization, and peripheral nerve regeneration.

[0008] LRG1 has also been implicated in disease pathogenesis. Evidence points to LRG1 being a proangiogenic factor, that through local upregulation contributes to the formation of defective vessels. The expression of LRG1 is associated with a variety of malignancies, such as non-small-cell lung cancer (NSCLC), ovarian cancer, and bladder cancer, and is thought to promote tumour growth via the angiogenesis process. LRG1 has been indicated to promote neovascularization in mouse models of ocular disease by potentiating endothelial TGF-p / activin receptor-like kinase 1 (ALK1) signalling. Circulating LRG1 levels have also been reported to be raised in severe COVID-19 patients and in patients with vasculitis, where vascular damage is a primary feature.

[0009] LRG1 has also been implicated in fibrosis. LRG1 has been reported to promote fibrosis in several tissues (e.g. lung, kidney, dermal and ocular tissues). However, LRG1 has also been reported to protect against fibrosis. Reduced levels of LRG1 in fibroblasts following selective deletion of PPARp / 5, made the epidermis of mutant mice thicker and more susceptible to inflammation and dermal fibrosis. LRG1 has also been shown to prevent the activation of skin fibroblasts by inhibiting pro-fibrotic TGFp signalling. A similar protective role has been described in the heart, where LRG1 is constitutively expressed via PPARp / 5 in resident fibroblasts to counteract TGFp function and preserve tissue integrity. Accordingly, the role of LRG1 in fibrosis is not well understood, in view of the conflicting studies demonstrating both pro- and anti-fibrotic activities. LRG1 expression has also been observed to be increased in several inflammatory disorders such as Still’s disease, psoriasis, lupus nephritis, rheumatoid arthritis and vasculitis. However, the role of LRG1 in these conditions remains largely unclear.

[0010] LRG1 is known or speculated to be involved in modulation of many signalling pathways. LRG1 likely modifies cell behaviour both directly, by altering the cell transcriptome, and indirectly by interfering with intermediate steps of the signalling cascades. LRG1 has been mainly described as a modifier of the TGFp canonical pathway. LRG1 promotes pathogenic angiogenesis in endothelial cells through the ALK1-S mad 1 / 5 / 8 pathway. LRG1 binds directly to the TGF-p accessory receptor endoglin, which, in the presence of TGF-01 , results in promotion of the pro-angiogenic Smad1 / 5 / 8 signalling pathway. LRG1 may also modulate the ALK5-Smad2 / 3 arm to favour the formation of myofibroblasts and Th17 lymphocytes.

[0011] TGFp non-canonical signalling is also likely to mediate some LRG1-driven biological functions including neutrophil activation and wound healing via AKT. Additional transduction factors involved in LRG1 signalling include (i) EGFR which promotes pancreatic cancer cell malignancy through p38 / MAPK, dissemination of melanoma cells and cornea repair through STAT3; (ii) the IL-6 / STAT3 axis which modulates neutrophil chemotaxis; (iii) Wnt / pcatenin which, in the heart, inhibit fibroblast proliferation and migration.

[0012] Diseases characterised by fibrosis and / or inflammation are a growing concern. For example, age-related macular degeneration (AMD) is the leading cause of blindness in the elderly worldwide. The neovascular form of AMD (sometimes referred to as nAMD, nvAMD or ‘wet’ AMD) is characterized by disorganised growth of blood vessels from the choroid which may penetrate the retinal pigment epithelium (RPE) layer and proliferate in the subretinal space, especially in the macular region. As the choroidal neovascularization (CNV) involutes, the cytokines and growth factors produced by RPE cells shift towards transform growth factor (TGF)p and tissue inhibitors of metalloproteinases (TIMP). As a result, the fibrotic scar may form resulting in irreversible loss of the central vision. Anti-vascular endothelial growth factor (VEGF) therapy is the current standard of care. Despite being effective in repressing CNV and blood vessel leakage, around 40% of AMD patients do not respond well to anti-VEGF treatment. Subretinal fibrosis affects about 50% of nAMD patients despite undergoing anti-VEGF therapy. This subgroup of patients may not regain vision despite suppression of exudative activity. Currently, no drug in the market targets subretinal fibrosis in nAMD patients. There is an urgent need to develop a new treatment for nAMD patients, especially those with subretinal fibrosis and who do not respond well to current anti- VEGF drugs.

[0013] Summary

[0014] In a first aspect the present disclosure provides an antigen-binding molecule, optionally isolated, which binds to LRG1 . In some embodiments, the antigen-binding molecule contacts the LRRCT region of LRG1 . In some embodiments, the antigen-binding molecule contacts the region of LRG1 shown in SEQ ID NO:183, 190 or 191.

[0015] In some embodiments, the antigen-binding molecule is capable of inhibiting one or more functions of LRG1.

[0016] In some embodiments, the antigen-binding molecule comprises:

[0017] (a)

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

[0019] 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

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

[0021] 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

[0022] (b)

[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] (c)

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

[0029] HC-CDR1 having the amino acid sequence of SEQ ID NO:2 HC-CDR2 having the amino acid sequence of SEQ ID NO:32 HC-CDR3 having the amino acid sequence of SEQ ID NO:33; 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:37 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:38; or

[0032] (d)

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

[0034] HC-CDR1 having the amino acid sequence of SEQ ID NO:43 HC-CDR2 having the amino acid sequence of SEQ ID NO:44 HC-CDR3 having the amino acid sequence of SEQ ID NO:45; and

[0035] (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO:25

[0036] LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:48.

[0037] In some embodiments, the antigen-binding molecule comprises: a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:1 , 17, 31 , 42, 82, 103, 105, 107, 122, 125; 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, 24, 36, 47, 86, 91 , 95, 98, 112, 114, 118, 129.

[0038] In some embodiments, the antigen-binding molecule comprises:

[0039] (i) 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

[0040] (ii) 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

[0041] (iii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:31 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:36; or

[0042] (iv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:42; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:47; or

[0043] (v)

[0044] (vi) -

[0045] (vii) 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:86; or (viii) 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:91 ; or

[0046] (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:95; or

[0047] (x) 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:98; or

[0048] (xi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:86; or

[0049] (xii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:91 ; or

[0050] (xiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:95; or

[0051] (ixv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:98; or

[0052] (xv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:86; 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:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:91 ; or

[0053] (xvii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:95; or

[0054] (xviii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:98; or

[0055] (ixx) 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:112; or

[0056] (xx) 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

[0057] (xxi) 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:118; or

[0058] (xxii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:122; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:112; or

[0059] (xxiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:122; 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 (ixv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:122; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:118; or

[0060] (xv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:125; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:112; or

[0061] (xvi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:125; 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

[0062] (xvii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:125; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:118; or

[0063] (xviii) 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:129.

[0064] In some embodiments, the antigen-binding molecule binds to human LRG1 , mouse LRG1 , and / or cynomolgus monkey LRG1 .

[0065] In some embodiments, the antigen-binding molecule is a multispecific antigen-binding molecule.

[0066] In some embodiments, the antigen-binding molecule is conjugated to a drug moiety or a detectable moiety.

[0067] The present disclosure also provides a chimeric antigen receptor (CAR) comprising an antigen-binding molecule according to the present disclosure.

[0068] The present disclosure also provides a nucleic acid, or a plurality of nucleic acids, optionally isolated, encoding the antigen-binding molecule according to the present disclosure.

[0069] The present disclosure also provides an expression vector, or a plurality of expression vectors, comprising a nucleic acid or a plurality of nucleic acids according to the present disclosure. The present disclosure also provides a cell comprising an antigen-binding molecule, a CAR, a nucleic acid or a plurality of nucleic, or an expression vector or a plurality of expression vectors according to the present disclosure.

[0070] The present disclosure also provides a method comprising culturing a cell according to the present disclosure under conditions suitable for expression of an antigen-binding molecule or CAR by the cell.

[0071] The present disclosure also provides a composition comprising the antigen-binding molecule, a CAR, a nucleic acid or a plurality of nucleic acids, an expression vector or a plurality of expression vectors, or a cell according to the present disclosure, and a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0072] The present disclosure provides an antigen-binding molecule, a CAR, a nucleic acid or a plurality of nucleic acids, an expression vector or a plurality of expression vectors, a cell, or composition according to the present disclosure, for use as a medicament.

[0073] The present disclosure also provides an antigen-binding molecule, a CAR, a nucleic acid or a plurality of nucleic acids, an expression vector or a plurality of expression vectors, a cell, or composition according to the present disclosure, for use in the treatment or prevention of a disease or condition in which LRG1 is pathologically-implicated.

[0074] The present disclosure also provides the use of an antigen-binding molecule, a CAR, a nucleic acid or a plurality of nucleic acids, an expression vector or a plurality of expression vectors, a cell, or composition according to the present disclosure, in the manufacture of a medicament for use in the treatment or prevention of a disease or condition in which LRG1 is pathologically-implicated.

[0075] The present disclosure also provides a method of treating or preventing a disease or condition in which LRG1 is pathologically-implicated, comprising administering to a subject a therapeutically- or prophylactically-effective amount of an antigen-binding molecule, a CAR, a nucleic acid or a plurality of nucleic acids, an expression vector or a plurality of expression vectors, a cell, or composition according to the present disclosure.

[0076] In some embodiments, the disease or condition is characterised by one or more of: fibrosis, inflammation and pathological angiogenesis.

[0077] In some embodiments, the disease or condition is characterised by: (i) fibrosis, inflammation and / or pathological angiogenesis of the eye; or (ii) fibrosis, inflammation and / or pathological angiogenesis of the pancreas. In some embodiments, the disease or condition is characterised by: (i) fibrosis, inflammation and / or pathological angiogenesis of the eye; (ii) fibrosis, inflammation and / or pathological angiogenesis of the pancreas; (iii) fibrosis and / or inflammation of the joints; (iv) fibrosis and / or inflammation of the skin; (v) fibrosis and / or inflammation of the bowel; or (vi) fibrosis and / or inflammation of the kidney; (vii) fibrosis and / or inflammation of the lungs.

[0078] In some embodiments, the disease or condition is selected from: macular degeneration, Age-related Macular Degeneration (AMD), Geographic Atrophy (‘dry’ or non-exudative AMD), early AMD, early onset macular degeneration (EOMD), intermediate AMD, late / advanced AMD, ‘wet’ (neovascular or exudative) AMD, choroidal neovascularisation (CNV), retinal dystrophy, glaucoma (open-angle or closed-angle), neuromyelitis optica (neuromyelitis optica spectrum disorder (NMOSD)), polypoidal choroidal vasculopathy, proliferative vitreoretinopathy (PVR), macular edema, drusen formation, Grave's ophthalmopathy, corneal opacification, subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis {e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis preretinal fibrosis, interlobular fibrosis, periductal fibrosis, diffuse interlobular fibrosis, diffuse intralobular fibrosis, pancreatitis, cystic fibrosis or pancreatic cancer. In some embodiments, the disease or condition is selected from: macular degeneration, Age-related Macular Degeneration (AMD), Geographic Atrophy (‘dry’ or non-exudative AMD), early AMD, early onset macular degeneration (EOMD), intermediate AMD, late / advanced AMD, ‘wet’ (neovascular or exudative) AMD, choroidal neovascularisation (CNV), retinal dystrophy, glaucoma (open-angle or closed-angle), neuromyelitis optica (neuromyelitis optica spectrum disorder (NMOSD)), diabetic macular edema (DME), diabetic retinopathy, proliferative diabetic retinopathy (PDR), polypoidal choroidal vasculopathy, proliferative vitreoretinopathy (PVR), macular edema, drusen formation, Grave's ophthalmopathy, corneal opacification, subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis {e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis preretinal fibrosis, interlobular fibrosis, periductal fibrosis, diffuse interlobular fibrosis, diffuse intralobular fibrosis, pancreatitis, cystic fibrosis, pancreatic cancer, skin fibrosis, kidney fibrosis, pulmonary fibrosis, rheumatoid arthritis, inflammatory bowel disease, diabetic nephropathy, or a diabetic wound.

[0079] In some embodiments, the disease or condition is cancer. In some embodiments, the cancer is selected from: a cancer comprising cells expressing / overexpressing LRG1 , a solid tumor, a metastatic tumor, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, liver cancer, hepatocellular carcinoma, skin cancer, melanoma, lung cancer, lung adenocarcinoma, squamous cell lung carcinoma, non-small cell lung cancer, breast cancer, breast carcinoma, ductal carcinoma, colorectal cancer, colorectal carcinoma, colorectal adenocarcinoma, gastric cancer, gastric carcinoma, gastric adenocarcinoma, head and neck cancer, squamous cell carcinoma of the head and neck (SCCHN), ovarian cancer, ovarian carcinoma, ovarian serous adenocarcinoma, kidney cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, renal papillary cell carcinoma, cervical cancer, cervical squamous cell carcinoma, esophageal cancer, esophageal adenocarcinoma, cholangiocarcinoma, uterine cancer, uterine corpus endometrial carcinoma, thyroid cancer, thyroid carcinoma, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma and thymoma. The present disclosure also provides, an in vitro complex, optionally isolated, comprising an antigenbinding molecule according to the present disclosure bound to LRG1 .

[0080] The present disclosure also provides a method for detecting LRG1 in a sample, comprising contacting a sample containing, or suspected to contain, LRG1 with an antigen-binding molecule according to the present disclosure, and detecting the formation of a complex of the antigen-binding molecule with LRG1 .

[0081] The present disclosure also provides a method of selecting or stratifying a subject for treatment with an LRG1 -targeted agent, the method comprising contacting, in vitro, a sample from the subject with an antigen-binding molecule according to the present disclosure and detecting the formation of a complex of the antigen-binding molecule with LRG1 .

[0082] The present disclosure also provides the use of an antigen-binding molecule according to the present disclosure as an in vitro or in vivo diagnostic or prognostic agent.

[0083] Description

[0084] The present invention is based on the inventors’ unexpected finding that a group of novel LRG1 -specific antigen-binding molecules are able to inhibit profibrotic, pro-angiogenic, pro-tumourigenic and / or pro- inflammatory functions of LRG1 . The present disclosure provides antigen-binding molecules that bind to LRG1 , having novel biophysical and / or functional properties as compared to antigen-binding molecules disclosed in the prior art.

[0085] LRG1

[0086] The present disclosure relates to LRG1 -specific antigen-binding molecules.

[0087] Leucine-rich a-2 glycoprotein 1 (LRG1) is a secreted member of the family of leucine-rich repeat proteins.

[0088] Human LRG1 is the protein identified by UniProt P02750 (SEQ ID NO:172). Human LRG1 comprises an N-terminal signal peptide (SEQ ID NO:174), 8 leucine-rich repeats (LRRs) (SEQ ID NO:175, 176, 177, 178, 179, 180, 181 , and 182) and a leucine-rich C-terminal domain (LRRCT) (SEQ ID NO:183). Human LRG1 contains 5 glycosylation sites (T37, N79, N186, N269 and N325) and two disulfide bonds (amino acid 43 to amino acid 56, and amino acid 303 to amino acid 329) (numbering is with reference to SEQ ID NO:172). Upon cleavage of the N-terminal signal peptide, LRG1 is released in the extracellular space. The mature form of human LRG1 is shown in SEQ ID NOU 73.

[0089] In this specification ‘LRG1 ’ refers to LRG1 from any species, and includes isoforms, fragments, variants or homologues from any species. In some embodiments LRG1 is LRG1 from a mammal (e.g. a therian, placental, epitherian, preptotheria, archontan, primate (rhesus, cynomolgous, non-human primate or human)). In some embodiments, the LRG1 is human LRG1 or mouse LRG1 . As used herein, a ‘fragment’, ‘variant’, ‘isoform’ or ‘homologue’ of a given protein may optionally be characterised as having at least 60% {e.g. one of >60%, >65%, >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91%, >92%, >93%, >94%, >95%, >96%, >97%, >98% or >99%) amino acid sequence identity to the amino acid sequence of the reference protein {e.g. a reference isoform).

[0090] 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. 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 LRG1 (UniProt P02750, v2; SEQ ID NO:172) and mouse LRG1 (UniProt: Q91XL1 , v1 ; SEQ ID NO:184) are homologues of one another. Homologues include orthologues.

[0091] Isoforms, fragments, variants or homologues of LRG1 according to the present disclosure may optionally be characterised as having at least 60% e.g. one of >60%, >65%, >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 the amino acid sequence of an immature or mature LRG1 isoform from a given species, e.g. human.

[0092] Isoforms, fragments, variants or homologues may optionally be functional isoforms, fragments, variants or homologues, e.g. having a functional property / activity of the reference LRG1 {e.g. human LRG1 ), as determined by analysis by a suitable assay for the functional property / activity. For example, an isoform, fragment, variant or homologue of LRG1 may display binding to and / or an association with a TGF-beta- receptor-ll / endoglin / ALK1 complex and / or a TGF-beta-receptor-ll / ALK5 complex.

[0093] In some embodiments, the LRG1 comprises, or consists of, an amino acid sequence having at least 70%, {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:172 or 173.

[0094] In some embodiments, the LRG1 comprises, or consists of, an amino acid sequence having at least 70% {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:184 or 185.

[0095] In some embodiments, the LRG1 comprises, or consists of, an amino acid sequence having at least 70% {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:188.

[0096] 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.

[0097] A fragment of LRG1 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.

[0098] In some embodiments, a fragment of LRG1 comprises, or consists of, an amino acid sequence having at least 70% {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:173, 174, 175, 176, 177, 178, 179, 180, 181 , 182, or 183.

[0099] In some embodiments, a fragment of LRG1 comprises, or consists of, an amino acid sequence having at least 70% {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:185, 186, or 187. In some embodiments, a fragment of LRG1 comprises, or consists of, an amino acid sequence having at least 70% {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:189.

[0100] As used herein, ‘LRRCT region’ or ‘leucine-rich C-terminal domain’ refers to an amino acid sequence having at least 70% {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:183, 187 or 189. In some embodiments, the LRRCT region has an amino acid sequence having at least 70% {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:183.

[0101] The structure and function of LRG1 is reviewed e.g. in Camilli et al. Journal of Biomedical Science (2022) 29(1 ):6, which is hereby incorporated by reference in its entirety.

[0102] Antigen-binding molecules

[0103] The present disclosure provides antigen-binding molecules capable of binding to LRG1 . An antigenbinding molecule that is capable of binding to a given target antigen may also be described as an antigenbinding molecule that binds to the given target antigen.

[0104] An ‘antigen-binding molecule’ refers to a molecule that binds to a given target antigen. Antigen-binding molecules include antibodies {i.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 molecule may be an antibody or an antigen-binding fragment thereof.

[0105] Antigen-binding molecules according to the present disclosure also include antibody-derived molecules, e.g. molecules comprising an antigen-binding region / domain derived from an antibody. Antibody-derived antigen-binding molecules may comprise an antigen-binding region / domain that comprises, or consists of, the antigen-binding region of an antibody {e.g. an antigen-binding fragment of an antibody). In some embodiments, the antigen-binding region / domain of an antibody-derived antigen-binding molecule may be or comprise the Fv {e.g. provided as an scFv) or the Fab region of an antibody, or the whole antibody. For example, antigen-binding molecules according to the present disclosure include antibody-drug conjugates (ADCs) comprising a (cytotoxic) drug moiety {e.g. as described hereinbelow). Antigen-binding molecules according to the present disclosure also include multispecific antigen-binding molecules such as immune cell engager molecules comprising a domain for recruiting (effector) immune cells (reviewed e.g. in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17: 418-434 and Ellerman, Methods (2019) 154:102-117, both of which are hereby incorporated by reference in their entirety), including BiTEs, BiKEs and TriKEs. Antigen-binding molecules according to the present disclosure also include chimeric antigen receptors (CARs), which are recombinant receptors providing both antigen-binding and T cell activating functions (CAR structure, function and engineering is reviewed e.g. in Dotti et al., Immunol Rev (2014) 257(1 ) and Jayaraman et al., EBioMedicine (2020) 58:102931 , both of which are hereby incorporated by reference in their entirety).

[0106] The antigen-binding molecule of the present disclosure comprises a moiety or moieties capable of binding to a target antigen(s). 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. In some embodiments, the moiety capable of binding to a target antigen comprises or consists of an aptamer capable of binding to the target antigen, e.g. a nucleic acid aptamer (reviewed, for example, in Zhou and Rossi Nat Rev Drug Discov. 2017 16(3) :181 -202). In some embodiments, the moiety capable of binding to a target antigen comprises or consists of an antigen-binding peptide / polypeptide, e.g. a peptide aptamer, thioredoxin, monobody, anticalin, Kunitz domain, avimer, knottin, fynomer, atrimer, DARPin, affibody, nanobody {i.e. a singledomain antibody (sdAb)), affilin, armadillo repeat protein (ArmRP), OBody or fibronectin - reviewed e.g. in Reverdatto et al., Curr Top Med Chem. 2015; 15(12): 1082-1101 , which is hereby incorporated by reference in its entirety (see also e.g. Boersma et al., J Biol Chem (2011 ) 286:41273-85 and Emanuel et al., Mabs (2011 ) 3:38-48).

[0107] 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. The antigen-binding molecules of the present disclosure generally comprise an antigen-binding domain comprising a VH and a VL of an antibody capable of specific binding to the target antigen. The antigenbinding domain formed by a VH and a VL may also be referred to herein as an Fv region.

[0108] An antigen-binding molecule may be, or may comprise, an antigen-binding polypeptide, or an antigenbinding polypeptide complex. An antigen-binding molecule may comprise more than one polypeptide which together form an antigen-binding domain. 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).

[0109] An antigen-binding molecule 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 molecule comprising two heavy chain polypeptides and two light chain polypeptides.

[0110] The antigen-binding molecules of the present disclosure may be designed and prepared using the sequences of monoclonal antibodies (mAbs), e.g. monoclonal antibodies capable of binding to LRG1 . 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.

[0111] 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.

[0112] 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.

[0113] 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. In some embodiments, the antigen-binding molecule comprises the CDRs of an antigen-binding molecule that binds to LRG1 . In some embodiments, the antigen-binding molecule comprises the FRs of an antigen-binding molecule that binds to LRG1 . In some embodiments, the antigen-binding molecule comprises the CDRs and the FRs of an antigen-binding molecule that binds to LRG1 . That is, in some embodiments, the antigen-binding molecule comprises the VH region and the VL region of an antigenbinding molecule that binds to LRG1 .

[0114] In some embodiments, the antigen-binding molecule comprises the CDRs, FRs and / or the VH and / or VL regions of a LRG1 -binding antibody clone described herein, or CDRs, FRs and / or VH and / or VL regions which are derived from those of a LRG1 -binding antibody clone described herein.

[0115] In some embodiments, a LRG1 -binding antibody clone is selected from: EBC58, EBC59, EBC60, EBC61 , EBC1 191 , EBC1 192, EBC1 193, EBC1 194, EBC1 195, EBC1 196, EBC1 197, EBC1 198, EBC1 199, EBC1200, EBC1201 , EBC1202, EBC1203, EBC1204, EBC1205, EBC1206, EBC1207, EBC1208, EBC1209, EBC1210, EBC121 1 and EBC1212.

[0116] In some embodiments, a LRG1 -binding antibody clone is selected from: EBC58, EBC59, EBC60, EBC61 , EBC1 191 , EBC1 192, EBC1 193, EBC1 194, EBC1 195, EBC1 196, EBC1 197, EBC1 198, EBC1 199, EBC1200, EBC1201 , EBC1202, EBC1206, EBC1207, EBC1208 and EBC1212.

[0117] In some embodiments, a LRG1 -binding antibody clone is selected from: EBC59, EBC60, EBC61 , EBC1 191 , EBC1 192, EBC1 195, EBC1 196, EBC1 199, EBC1200 and EBC1212.

[0118] In some embodiments, the antigen-binding molecule comprises a VH region according to one of (1 ) to (6) below:

[0119] (1 ) a VH region incorporating the following CDRs:

[0120] HC-CDR1 having the amino acid sequence of SEQ ID NO:2

[0121] HC-CDR2 having the amino acid sequence of SEQ ID NO:3

[0122] 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.

[0123] (2) a VH region incorporating the following CDRs:

[0124] HC-CDR1 having the amino acid sequence of SEQ ID NO:18

[0125] HC-CDR2 having the amino acid sequence of SEQ ID NO:19

[0126] 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. (3) a VH region incorporating the following CDRs:

[0127] HC-CDR1 having the amino acid sequence of SEQ ID NO:2

[0128] HC-CDR2 having the amino acid sequence of SEQ ID NO:32

[0129] HC-CDR3 having the amino acid sequence of SEQ ID NO:33, 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.

[0130] (4) a VH region incorporating the following CDRs:

[0131] HC-CDR1 having the amino acid sequence of SEQ ID NO:43

[0132] HC-CDR2 having the amino acid sequence of SEQ ID NO:44

[0133] HC-CDR3 having the amino acid sequence of SEQ ID NO:45, 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.

[0134] (5) -

[0135] (6) -

[0136] In some embodiments, the antigen-binding molecule comprises a VH region according to one of (7) to (19) below:

[0137] (7) a VH region incorporating the following FRs:

[0138] HC-FR1 having the amino acid sequence of SEQ ID NO:5

[0139] HC-FR2 having the amino acid sequence of SEQ ID NO:6

[0140] HC-FR3 having the amino acid sequence of SEQ ID NO:7

[0141] 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.

[0142] (8) a VH region incorporating the following FRs:

[0143] HC-FR1 having the amino acid sequence of SEQ ID NO:21

[0144] HC-FR2 having the amino acid sequence of SEQ ID NO:22

[0145] HC-FR3 having the amino acid sequence of SEQ ID NO:23

[0146] 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. (9) a VH region incorporating the following FRs:

[0147] HC-FR1 having the amino acid sequence of SEQ ID NO:34

[0148] HC-FR2 having the amino acid sequence of SEQ ID NO:35

[0149] HC-FR3 having the amino acid sequence of SEQ ID NO:23

[0150] 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.

[0151] (10) a VH region incorporating the following FRs:

[0152] HC-FR1 having the amino acid sequence of SEQ ID NO:46

[0153] HC-FR2 having the amino acid sequence of SEQ ID NO:35

[0154] HC-FR3 having the amino acid sequence of SEQ ID NO:23

[0155] 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.

[0156] (11 ) -

[0157] (12) -

[0158] (13) a VH region incorporating the following FRs:

[0159] HC-FR1 having the amino acid sequence of SEQ ID NO:83

[0160] HC-FR2 having the amino acid sequence of SEQ ID NO:84

[0161] HC-FR3 having the amino acid sequence of SEQ ID NO:85

[0162] 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.

[0163] (14) a VH region incorporating the following FRs:

[0164] HC-FR1 having the amino acid sequence of SEQ ID NO:83

[0165] HC-FR2 having the amino acid sequence of SEQ ID NO:84

[0166] HC-FR3 having the amino acid sequence of SEQ ID NQ:104

[0167] 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.

[0168] (15) a VH region incorporating the following FRs: HC-FR1 having the amino acid sequence of SEQ ID NO:83

[0169] HC-FR2 having the amino acid sequence of SEQ ID NO:84

[0170] HC-FR3 having the amino acid sequence of SEQ ID NQ:106

[0171] 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.

[0172] (16) a VH region incorporating the following FRs:

[0173] HC-FR1 having the amino acid sequence of SEQ ID NQ:108

[0174] HC-FR2 having the amino acid sequence of SEQ ID NQ:109

[0175] HC-FR3 having the amino acid sequence of SEQ ID NO:1 10

[0176] HC-FR4 having the amino acid sequence of SEQ ID NO:1 1 1 , 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.

[0177] (17) a VH region incorporating the following FRs:

[0178] HC-FR1 having the amino acid sequence of SEQ ID NO:123

[0179] HC-FR2 having the amino acid sequence of SEQ ID NO:84

[0180] HC-FR3 having the amino acid sequence of SEQ ID NO:124

[0181] HC-FR4 having the amino acid sequence of SEQ ID NO:1 1 1 , 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.

[0182] (18) a VH region incorporating the following FRs:

[0183] HC-FR1 having the amino acid sequence of SEQ ID NO:126

[0184] HC-FR2 having the amino acid sequence of SEQ ID NO:127

[0185] HC-FR3 having the amino acid sequence of SEQ ID NO:128

[0186] HC-FR4 having the amino acid sequence of SEQ ID NO:1 1 1 , 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.

[0187] (19) a VH region incorporating the following FRs:

[0188] HC-FR1 having the amino acid sequence of SEQ ID NO:83

[0189] HC-FR2 having the amino acid sequence of SEQ ID NO:84

[0190] HC-FR3 having the amino acid sequence of SEQ ID NO:85

[0191] 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.

[0192] In some embodiments, the antigen-binding molecule comprises a VH region comprising the CDRs according to any one of (1 ) to (6) above, and the FRs according to any one of (7) to (19) above.

[0193] In some embodiments, the antigen-binding molecule comprises a VH region according to one of (20) to (32) below:

[0194] (20) a VH region comprising the CDRs according to (1 ) and the FRs according to (7).

[0195] (21 ) a VH region comprising the CDRs according to (2) and the FRs according to (8).

[0196] (22) a VH region comprising the CDRs according to (3) and the FRs according to (9).

[0197] (23) a VH region comprising the CDRs according to (4) and the FRs according to (10).

[0198] (24) -

[0199] (25) -

[0200] (26) a VH region comprising the CDRs according to (2) and the FRs according to (13).

[0201] (27) a VH region comprising the CDRs according to (2) and the FRs according to (14).

[0202] (28) a VH region comprising the CDRs according to (2) and the FRs according to (15).

[0203] (29) a VH region comprising the CDRs according to (2) and the FRs according to (16).

[0204] (30) a VH region comprising the CDRs according to (2) and the FRs according to (17).

[0205] (31 ) a VH region comprising the CDRs according to (2) and the FRs according to (18).

[0206] (32) a VH region comprising the CDRs according to (2) and the FRs according to (19).

[0207] In some embodiments, the antigen-binding molecule comprises a VH region according to one of (33) to (44) below:

[0208] (33) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:1 . (34) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity)to the amino acid sequence of SEQ ID NO:17.

[0209] (35) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:31 .

[0210] (36) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:42.

[0211] (37) -

[0212] (38) -

[0213] (39) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:82.

[0214] (40) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NQ:103.

[0215] (41 ) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NQ:105.

[0216] (42) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NQ:107.

[0217] (43) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:122.

[0218] (44) a VH region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:125. In some embodiments, the antigen-binding molecule comprises a VL region according to one of (45) to

[0219] (50) below:

[0220] (45) a VL region incorporating the following CDRs:

[0221] LC-CDR1 having the amino acid sequence of SEQ ID NO:10

[0222] LC-CDR2 having the amino acid sequence of SEQ ID NO:1 1

[0223] 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.

[0224] (46) a VL region incorporating the following CDRs:

[0225] LC-CDR1 having the amino acid sequence of SEQ ID NO:25

[0226] LC-CDR2 having the amino acid sequence of SEQ ID NO:26

[0227] 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.

[0228] (47) a VL region incorporating the following CDRs:

[0229] LC-CDR1 having the amino acid sequence of SEQ ID NO:37

[0230] LC-CDR2 having the amino acid sequence of SEQ ID NO:26

[0231] LC-CDR3 having the amino acid sequence of SEQ ID NO:38, 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.

[0232] (48) a VL region incorporating the following CDRs:

[0233] LC-CDR1 having the amino acid sequence of SEQ ID NO:25

[0234] LC-CDR2 having the amino acid sequence of SEQ ID NO:26

[0235] LC-CDR3 having the amino acid sequence of SEQ ID NO:48, 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.

[0236] (49) -

[0237] (50) -

[0238] In some embodiments, the antigen-binding molecule comprises a VL region according to one of (51 ) to

[0239] (64) below:

[0240] (51 ) a VL region incorporating the following FRs:

[0241] LC-FR1 having the amino acid sequence of SEQ ID NO:13

[0242] LC-FR2 having the amino acid sequence of SEQ ID NO:14

[0243] LC-FR3 having the amino acid sequence of SEQ ID NO:15

[0244] 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.

[0245] (52) a VL region incorporating the following FRs:

[0246] LC-FR1 having the amino acid sequence of SEQ ID NO:28

[0247] LC-FR2 having the amino acid sequence of SEQ ID NO:29

[0248] LC-FR3 having the amino acid sequence of SEQ ID NQ:30

[0249] 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.

[0250] (53) a VL region incorporating the following FRs:

[0251] LC-FR1 having the amino acid sequence of SEQ ID NO:39

[0252] LC-FR2 having the amino acid sequence of SEQ ID NQ:40

[0253] LC-FR3 having the amino acid sequence of SEQ ID NO:41

[0254] 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.

[0255] (54) a VL region incorporating the following FRs:

[0256] LC-FR1 having the amino acid sequence of SEQ ID NO:49

[0257] LC-FR2 having the amino acid sequence of SEQ ID NQ:50

[0258] LC-FR3 having the amino acid sequence of SEQ ID NO:51

[0259] 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.

[0260] (55) -

[0261] (56) -

[0262] (57) a VL region incorporating the following FRs:

[0263] LC-FR1 having the amino acid sequence of SEQ ID NO:87

[0264] LC-FR2 having the amino acid sequence of SEQ ID NO:88

[0265] LC-FR3 having the amino acid sequence of SEQ ID NO:89

[0266] LC-FR4 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-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.

[0267] (58) a VL region incorporating the following FRs:

[0268] LC-FR1 having the amino acid sequence of SEQ ID NO:92

[0269] LC-FR2 having the amino acid sequence of SEQ ID NO:93

[0270] LC-FR3 having the amino acid sequence of SEQ ID NO:94

[0271] LC-FR4 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-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.

[0272] (59) a VL region incorporating the following FRs:

[0273] LC-FR1 having the amino acid sequence of SEQ ID NO:96

[0274] LC-FR2 having the amino acid sequence of SEQ ID NO:93

[0275] LC-FR3 having the amino acid sequence of SEQ ID NO:97

[0276] LC-FR4 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-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.

[0277] (60) a VL region incorporating the following FRs:

[0278] LC-FR1 having the amino acid sequence of SEQ ID NO:99

[0279] LC-FR2 having the amino acid sequence of SEQ ID NQ:100

[0280] LC-FR3 having the amino acid sequence of SEQ ID NQ:101

[0281] LC-FR4 having the amino acid sequence of SEQ ID NQ:102, 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.

[0282] (61 ) a VL region incorporating the following FRs:

[0283] LC-FR1 having the amino acid sequence of SEQ ID NO:1 13

[0284] LC-FR2 having the amino acid sequence of SEQ ID NO:88

[0285] LC-FR3 having the amino acid sequence of SEQ ID NO:89

[0286] LC-FR4 having the amino acid sequence of SEQ ID NQ:102, 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.

[0287] (62) a VL region incorporating the following FRs: LC-FR1 having the amino acid sequence of SEQ ID NO:1 15

[0288] LC-FR2 having the amino acid sequence of SEQ ID NO:1 16

[0289] LC-FR3 having the amino acid sequence of SEQ ID NO:1 17

[0290] LC-FR4 having the amino acid sequence of SEQ ID NQ:102, 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.

[0291] (63) a VL region incorporating the following FRs:

[0292] LC-FR1 having the amino acid sequence of SEQ ID NO:1 19

[0293] LC-FR2 having the amino acid sequence of SEQ ID NQ:120

[0294] LC-FR3 having the amino acid sequence of SEQ ID NO:121

[0295] LC-FR4 having the amino acid sequence of SEQ ID NQ:102, 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.

[0296] (64) a VL region incorporating the following FRs:

[0297] LC-FR1 having the amino acid sequence of SEQ ID NO:99

[0298] LC-FR2 having the amino acid sequence of SEQ ID NQ:130

[0299] LC-FR3 having the amino acid sequence of SEQ ID NO:131

[0300] LC-FR4 having the amino acid sequence of SEQ ID NQ:102, 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.

[0301] In some embodiments, the antigen-binding molecule comprises a VL region comprising the CDRs according to any one of (45) to (50) above, and the FRs according to any one of (51 ) to (64) above.

[0302] In some embodiments, the antigen-binding molecule comprises a VL region according to one of (65) to (92) below:

[0303] (65) a VL region comprising the CDRs according to (45) and the FRs according to (51 ).

[0304] (66) a VL region comprising the CDRs according to (46) and the FRs according to (52).

[0305] (67) a VL region comprising the CDRs according to (47) and the FRs according to (53).

[0306] (68) a VL region comprising the CDRs according to (48) and the FRs according to (54).

[0307] (69) - (70) -

[0308] (71 ) a VL region comprising the CDRs according to (46) and the FRs according to (57).

[0309] (72) a VL region comprising the CDRs according to (46) and the FRs according to (58).

[0310] (73) a VL region comprising the CDRs according to (46) and the FRs according to (59).

[0311] (74) a VL region comprising the CDRs according to (46) and the FRs according to (60).

[0312] (75) a VL region comprising the CDRs according to (46) and the FRs according to (61 ).

[0313] (76) a VL region comprising the CDRs according to (46) and the FRs according to (62).

[0314] (77) a VL region comprising the CDRs according to (46) and the FRs according to (63).

[0315] (78) a VL region comprising the CDRs according to (46) and the FRs according to (64).

[0316] In some embodiments, the antigen-binding molecule comprises a VL region according to one of (79) to

[0317] (92) below:

[0318] (79) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:9.

[0319] (80) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:24.

[0320] (81 ) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:36.

[0321] (82) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:47.

[0322] (83) -

[0323] (84) - (85) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:86.

[0324] (86) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:91 .

[0325] (87) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:95.

[0326] (88) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:98.

[0327] (89) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:112.

[0328] (90) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:114.

[0329] (91 ) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:118.

[0330] (92) a VL region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:129.

[0331] In some embodiments, the antigen-binding molecule comprises a VH region according to any one of (1 ) to (44) above, and a VL region according to any one of (45) to (92) above.

[0332] In embodiments in accordance with the present disclosure, one or more amino acids 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 {i.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 a!., Meth. Enzym. 202 (1991 ) 301 -336.

[0333] 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:

[0334] 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.

[0335] 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: 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.

[0336] 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.

[0337] The VH and VL region of an antigen-binding region of an antibody together constitute the Fv region. In some embodiments, the antigen-binding molecule according to the present disclosure comprises, or consists of, an Fv region that binds to LRG1 . 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).

[0338] 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 molecule 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).

[0339] In some embodiments, the antigen-binding molecule described herein comprises, or consists of, a whole antibody that binds to LRG1 . 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.

[0340] 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. IgG 1 , lgG2, lgG3, lgG4), IgA (e.g. lgA1 , lgA2), IgD, IgE, or IgM. The light chain may be kappa (K) or lambda (A).

[0341] Herein, a ‘CH1 domain’ refers to an amino acid sequence corresponding to the CH1 domain of an immunoglobulin (Ig). The CH1 domain is the region of an Ig formed by positions 118 to 215 of the immunoglobulin constant domain, according to the EU numbering system (described in Edelman et al., Proc Natl Acad Sci USA (1969) 63(1 ): 78-85). A ‘hinge domain’ refers to an amino acid sequence corresponding to the hinge domain of an Ig. The hinge domain is the region of an Ig formed by positions 216 to 230 of the immunoglobulin constant domain, according to the EU numbering system. A ‘CH2 domain’ refers to an amino acid sequence corresponding to the CH2 domain of an Ig. The CH2 domain is the region of an Ig formed by positions 231 to 340 of the immunoglobulin constant domain, according to the EU numbering system. A ‘CH3 domain’ refers to an amino acid sequence corresponding to the CH3 domain of an immunoglobulin (Ig). The CH3 domain is the region of an Ig formed by positions 341 to 447 of the immunoglobulin constant domain, according to the EU numbering system. A ‘CH2-CH3 region’ refers to an amino acid sequence corresponding to the CH2 and CH3 domains of an immunoglobulin (Ig). The CH2-CH3 region is the region of an Ig formed by positions 231 to 447 of the immunoglobulin constant domain, according to the EU numbering system.

[0342] In some embodiments, the antigen-binding molecule described herein comprises, or consists of, an IgG (e.g. IgG 1 , lgG2, lgG3, lgG4), IgA (e.g. lgA1 , lgA2), IgD, IgE, or IgM that binds to LRG1 .

[0343] In some embodiments, the antigen-binding molecule 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. Ig A1 , Ig A2) , IgD, IgE or IgM, e.g. a human IgG (e.g. hlgG 1 , hlgG2, hlgG3, hlgG4), hlgA (e.g. hlgA1 , hlgA2), hlg D, h Ig E 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 ml , G1 m2, G1 m3 or G1 ml 7).

[0344] In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NOU 32 or 137. In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NOU 93. In some preferred embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NOU 32. In some embodiments, the antigen-binding molecule comprises a CH1 region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:133. In some embodiments, the antigen-binding molecule comprises a CH2 region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:135. In some embodiments, the antigen-binding molecule comprises a CH3 region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:136.

[0345] In some embodiments, the antigen-binding molecule comprises a hinge region comprising an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:134.

[0346] 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 molecule as described herein.

[0347] In some embodiments, the antigen-binding molecule 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.

[0348] In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:138, 139, 140, 141 , 142, 143. In some embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:194. In some preferred embodiments, the antigen-binding molecule comprises an amino acid sequence having at least 70% 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% sequence identity) to the amino acid sequence of SEQ ID NO:138

[0349] In some embodiments, the antigen-binding molecule is or comprises a monoclonal antibody, or an antigen-binding fragment thereof. In some embodiments, the antigen-binding molecule 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 disulf ide-linked 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.

[0350] In some embodiments, the antigen-binding molecule 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.

[0351] In some embodiments, the antigen-binding molecule is a mouse / human chimeric antibody / antibody fragment (i.e. an antigen-binding molecule comprising mouse antibody variable domains and human antibody constant regions). In some embodiments, the antigen-binding molecule is a humanised antibody / antibody fragment. In some embodiments, the antigen-binding molecule comprises mouse antibody CDRs and human antibody framework and constant regions.

[0352] Mouse / human chimeric antigen-binding molecules 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.

[0353] Humanised antigen-binding molecules 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. Aspects of the present disclosure relate to multispecific antigen-binding molecules. By ‘multispecific’ it is meant that the antigen-binding molecule displays specific binding to more than one target. In some embodiments, the antigen-binding molecule is a bispecific antigen-binding molecule. In some embodiments, the antigen-binding molecule comprises at least two different antigen-binding domains {i.e. at least two antigen-binding domains, e.g. comprising non-identical VHs and VLs).

[0354] In some embodiments, the antigen-binding molecule binds to LRG1 and another target {e.g. an antigen other than LRG1 ), and so is at least bispecific. The term ‘bispecific’ means that the antigen-binding molecule is able to bind specifically to at least two distinct antigenic determinants.

[0355] It will be appreciated that an antigen-binding molecule according to the present disclosure {e.g. a multispecific antigen-binding molecule) may comprise antigen-binding molecules capable of binding to the targets for which the antigen-binding molecule is specific. For example, an antigen-binding molecule that binds to LRG1 and an antigen other than LRG1 may comprise: (i) an antigen-binding molecule that binds to LRG1 , and (ii) an antigen-binding molecule that binds to an antigen other than LRG1 .

[0356] It will also be appreciated that an antigen-binding molecule according to the present disclosure {e.g. a multispecific antigen-binding molecule) may comprise antigen-binding polypeptides or antigen-binding polypeptide complexes capable of binding to the targets for which the antigen-binding molecule is specific.

[0357] In some embodiments, a component antigen-binding molecule of a larger antigen-binding molecule {e.g. a multispecific antigen-binding molecule) may be referred to e.g. as an ‘antigen-binding domain’ or ‘antigen-binding region’ of the larger antigen-binding molecule.

[0358] Multispecific antigen-binding molecules 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. IgGa, F(ab’)2 or CovX-Body; IgG or IgG-like molecules, e.g. IgG, chimeric IgG, KA-body common HC; CH1 / CL fusion proteins, e.g. scFv2-CH1 / CL, VHH2-CH1 / CL; ‘variable domain only’ bispecific antigenbinding molecules, e.g. tandem scFv (taFV), triplebodies, diabodies (Db), dsDb, Db(kih), DART, scDB, dsFv-dsFv, tandAbs, triple heads, tandem dAb / VHH, tertravalent dAb.VHH; Non-lg fusion proteins, e.g. scFvz-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-Fabz-IgG-cytokinez; 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.

[0359] The skilled person is able to design and prepare bispecific antigen-binding molecules. Methods for producing multispecific antigen-binding molecules include chemically crosslinking antigen-binding molecules 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:l V: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.

[0360] Other methods for producing multispecific antigen-binding molecules 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.

[0361] Multispecific antigen-binding molecules according to the present disclosure can also be produced recombinantly, by expression from e.g. a nucleic acid construct encoding polypeptides for the antigenbinding molecules, 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 antigenbinding molecules, Methods Mol. Med. 2000; 40:333-339, the entire contents of both of which are hereby incorporated by reference.

[0362] For example, a DNA construct encoding the light and heavy chain variable domains for the two antigenbinding fragments (i.e. the light and heavy chain variable domains for the antigen-binding fragment capable of binding LRG1 , 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.

[0363] Fc regions

[0364] In some embodiments, the antigen-binding molecules of the present disclosure comprise an Fc region. In some embodiments, the antigen-binding molecules of the present disclosure does not comprise an Fc region.

[0365] 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.

[0366] 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 etal., 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 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.

[0367] 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 ).

[0368] 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.

[0369] 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.

[0370] 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.

[0371] 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, FcyRHb, FcyRHc, FcyRllla and FcyRHIb. In some embodiments, the Fc region comprises modification to increase binding to FcyRHIa. 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 FcyRHb. 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.

[0372] 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 etal., Mol Cancer Ther. (2008) 7:2517-2527.

[0373] 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. 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.

[0374] 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 etal. Biol Chem. (2014) 289:15309-15318.

[0375] In some embodiments, the Fc region comprises modification to reduce / prevent an Fc-mediated function. 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.

[0376] 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.

[0377] In some embodiments, the Fc region is not able to induce one or more Fc-mediated functions (i.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).

[0378] 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. 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.

[0379] 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 etal., 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 H268Q / V309L / A330S / P331 S 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 V234A / G237A / P238S / H268A / V309L / A330S / P331 S 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 / P331 S as described in US 2015 / 0044231 A1 .

[0380] 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 C1 q 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 IgG 1 . The combination of substitutions L234A / L235E / G237A / A330S / P331 S in lgG1 Fc is disclosed in US 2015 / 0044231 A1 to abolish induction of phagocytosis, ADCC and CDC.

[0381] 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).

[0382] 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.

[0383] In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235A / P329G.

[0384] In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235A / P329G / N297Q.

[0385] In some embodiments, the Fc region comprises modification corresponding to corresponding to the combination of substitutions L234A / L235E / G237A / A330S / P331 S.

[0386] In some embodiments, the Fc region comprises modification corresponding to corresponding to the substitution S228P, e.g. in lgG4.

[0387] In some embodiments - particularly embodiments in which the antigen-binding molecule 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.

[0388] In some embodiments, the antigen antigen-binding molecule of the present disclosure 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. Polypeptides and particular exemplary antigen-binding molecules

[0389] The present disclosure also provides polypeptide constituents of antigen-binding molecules. The polypeptides may be provided in isolated or substantially purified form.

[0390] The antigen-binding molecule of the present disclosure may be, or may comprise, a complex of polypeptides.

[0391] 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.

[0392] 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.

[0393] 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).

[0394] 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.

[0395] 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.

[0396] In some embodiments, the polypeptide according to the present disclosure comprises a structure from N- to C-terminus according to one of the following:

[0397] (i) VH

[0398] (ii) VL

[0399] (iii) VH-CH1

[0400] (iv) VL-CL

[0401] (v) VL-CH1

[0402] (vi) VH-CL

[0403] (vii) VH-CH1 -CH2-CH3

[0404] (viii) VL-CL-CH2-CH3

[0405] (ix) VL-CH1 -CH2-CH3 (x) VH-CL-CH2-CH3

[0406] Also provided by the present disclosure are antigen-binding molecules composed of the polypeptides of the present disclosure. In some embodiments, the antigen-binding molecule of the present disclosure comprises one of the following combinations of polypeptides:

[0407] (A) VH + VL

[0408] (B) VH-CH1 + VL-CL

[0409] (C) VL-CH1 + VH-CL

[0410] (D) VH-CH1 -CH2-CH3 + VL-CL

[0411] (E) VH-CL-CH2-CH3 + VL-CH1

[0412] (F) VL-CH1 -CH2-CH3 + VH-CL

[0413] (G) VL-CL-CH2-CH3 + VH-CH1

[0414] (H) VH-CH1 -CH2-CH3 + VL-CL-CH2-CH3

[0415] (I) VH-CL-CH2-CH3 + VL-CH1 -CH2-CH3

[0416] In some embodiments, the antigen-binding molecule 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 molecule comprises two polypeptides comprising the structure VH- CH1 -CH2-CH3, and two polypeptides comprising the structure VL-CL.

[0417] In some embodiments, the antigen-binding molecule of the present disclosure comprises one of the following combinations of polypeptides:

[0418] (J) VH (anti-LRG1) + VL (anti-LRG1)

[0419] (K) VH (anti-LRG1 )-CH1 + VL (anti-LRGI )-CL

[0420] (L) VL (anti-LRG1 )-CH1 + VH (anti-LRGI )-CL

[0421] (M) VH (anti-LRG1 )-CH1 -CH2-CH3 + VL (anti-LRGI )-CL

[0422] (N) VH (anti-LRG1 )-CL-CH2-CH3 + VL (anti-LRG1 )-CH1

[0423] (O) VL (anti-LRG1 )-CH1 -CH2-CH3 + VH (anti-LRGI )-CL

[0424] (P) VL (anti-LRG1 )-CL-CH2-CH3 + VH (anti-LRG1 )-CH1

[0425] (Q) VH (anti-LRG1 )-CH1 -CH2-CH3 + VL (anti-LRG1 )-CL-CH2-CH3

[0426] Wherein: ‘VH(anti-LRGI )’ refers to the VH of an antigen-binding molecule capable of binding to LRG1 as described herein, e.g. as defined in one of (1 ) to (44); and ‘VL(anti-LRGI )’ refers to the VL of an antigenbinding molecule capable of binding to LRG1 as described herein, e.g. as defined in one of (45) to (92).

[0427] In some embodiments, the antigen-binding molecule of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% (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:1 , 17, 31 , 42, 82, 103, 105, 107, 122, or 125. In some embodiments, the antigen-binding molecule of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% (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:9. 24, 36, 47, 86, 91 , 95, 98, 112, 114, 118, or 129.

[0428] In some embodiments, the antigen-binding molecule of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% (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:144, 145, 146, 147, 150, 151 , 152, 153, 154, or 155.

[0429] In some embodiments, the antigen-binding molecule of the present disclosure comprises a polypeptide which comprises or consists of an amino acid sequence having at least 70% (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:156, 157, 158, 159, 162, 163, 164, 165, 166, 167, 168, or 169.

[0430] In some embodiments, the antigen-binding molecule 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 EBC58, EBC59, EBC60, EBC61 , EBC1191 , EBC1192, EBC1193, EBC1194, EBC1195, EBC1196, EBC1197, EBC1198, EBC1199, EBC1200, EBC1201 , EBC1202, EBC1203, EBC1204, EBC1205, EBC1206, EBC1207, EBC1208, EBC1209, EBC1210, EBC1211 and EBC1212, as shown in Table A herein. That is, in some embodiments, the antigen-binding molecule 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.

[0431] In some embodiments, the antigen-binding molecule 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 EBC58, EBC59, EBC60, EBC61 , EBC1191 , EBC1192, EBC1193, EBC1194, EBC1195, EBC1196, EBC1197, EBC1198, EBC1199, EBC1200, EBC1201 , EBC1202, EBC1203, EBC1204, EBC1205, EBC1206, EBC1207, EBC1208, EBC1209, EBC1210, EBC1211 and EBC1212, as shown in Table B herein. That is, in some embodiments, the antigen-binding molecule 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. In some embodiments, the antigen-binding molecule of the present disclosure comprises a polypeptide or polypeptides comprising: (i) an amino acid sequence having at least 70% {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 an amino acid sequence indicated in column A of Table C, and (ii) an amino acid sequence having at least 70% {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 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.

[0432] In some embodiments, the antigen-binding molecule of the present disclosure comprises a polypeptide or polypeptides comprising a VH region and a VL region of a clone selected from EBC58, EBC59, EBC60, EBC61 , EBC1191 , EBC1192, EBC1193, EBC1194, EBC1195, EBC1196, EBC1197, EBC1198, EBC1199, EBC1200, EBC1201 , EBC1202, EBC1203, EBC1204, EBC1205, EBC1206, EBC1207, EBC1208, EBC1209, EBC1210, EBC1211 and EBC1212, as shown in Table C herein. That is, in some embodiments, the antigen-binding molecule 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.

[0433] In some embodiments, the antigen-binding molecule of the present disclosure comprises: (i) a polypeptide comprising or consisting of an amino acid sequence having at least 70% {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 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% {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 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.

[0434] In some embodiments, the antigen-binding molecule of the present disclosure comprises the polypeptides of an antigen-binding molecule according to any one of [1] to

[0028] as detailed in Table D herein. That is, in some embodiments, the antigen-binding molecule 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.

[0435] Linkers and additional sequences

[0436] In some embodiments, the antigen-binding molecules and polypeptides of the present disclosure comprise one or more linker 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. 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.

[0437] 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 41 1 ) 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.

[0438] The antigen-binding molecules and polypeptides of the present disclosure may additionally comprise further amino acids or sequences of amino acids. For example, the antigen-binding molecules and polypeptides may comprise amino acid sequence(s) to facilitate expression, folding, trafficking, processing, purification, or detection of the antigen-binding molecule / polypeptide. For example, antigenbinding molecules and polypeptides of the present disclosure may additionally comprise a sequence of amino acids forming a detectable moiety, e.g. as described hereinbelow.

[0439] The antigen-binding molecules and polypeptides of the present disclosure 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.

[0440] The signal peptide may be present at the N-terminus of the antigen-binding molecule / polypeptide, and may be present in the newly synthesised antigen-binding molecule / polypeptide. The signal peptide provides for efficient trafficking and secretion of the antigen-binding molecule / polypeptide. Signal peptides are often removed by cleavage, and thus are not comprised in the mature antigen-binding molecule / polypeptide secreted from the cell expressing the antigen-binding molecule / polypeptide.

[0441] 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., 201 1 Nature Methods 8: 785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24: 2172- 2176).

[0442] Labels and conjugates

[0443] In some embodiments, the antigen-binding molecules of the present disclosure additionally comprise a detectable moiety.

[0444] 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.

[0445] 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.

[0446] In some embodiments, the antigen-binding molecule / polypeptide 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.

[0447] In some embodiments, the antigen-binding molecule / polypeptide 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.

[0448] In some embodiments, the antigen-binding molecules of the present disclosure are conjugated to a chemical moiety. The chemical moiety may be a moiety for providing a therapeutic effect, i.e. a drug moiety. A drug moiety may be a small molecule {e.g. a low molecular weight (< 1000 daltons, typically between -300-700 daltons) organic compound). Drug moieties are described e.g. in Parslow et al., Biomedicines. 2016 Sep; 4(3):14 (hereby incorporated by reference in its entirety). In some embodiments, a drug moiety may be or comprise a cytotoxic agent. In some embodiments, a drug moiety may be or comprise a chemotherapeutic agent. Drug moieties include e.g. calicheamicin, DM1 , DM4, monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), SN-38, doxorubicin, duocarmycin, D6.5 and PBD.

[0449] Antigen-binding molecules according to the present disclosure also include antibody-derived molecules, e.g. molecules comprising an antigen-binding region / domain derived from an antibody. Antibody-derived antigen-binding molecules may comprise an antigen-binding region / domain that comprises, or consists of, the antigen-binding region of an antibody {e.g. an antigen-binding fragment of an antibody). In some embodiments, the antigen-binding region / domain of an antibody-derived antigen-binding molecule may be or comprise the Fv {e.g. provided as an scFv) or the Fab region of an antibody, or the whole antibody. For example, antigen-binding molecules according to the present disclosure include antibody-drug conjugates (ADCs) comprising a (cytotoxic) drug moiety. Antigen-binding molecules according to the present disclosure also include multispecific antigen-binding molecules such as immune cell engager molecules comprising a domain for recruiting (effector) immune cells (reviewed e.g. in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17: 418-434 and Ellerman, Methods (2019) 154:102-1 17, both of which are hereby incorporated by reference in their entirety), including BiTEs, BiKEs and TriKEs. Antigenbinding molecules according to the present disclosure also include chimeric antigen receptors (CARs), which are recombinant receptors providing both antigen-binding and T cell activating functions (CAR structure, function and engineering is reviewed e.g. in Dotti et al., Immunol Rev (2014) 257(1 ), which is hereby incorporated by reference in its entirety).

[0450] In some embodiments, an antigen-binding molecule according to the present disclosure comprises a drug moiety. The antigen-binding molecule 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). FDA approved ADCs currently on the market are described in Tong et al., Molecules. 2021 Oct; 26(19): 5847 (hereby incorporated by reference in its entirety).

[0451] In some embodiments the antibody-drug conjugate comprises an antigen binding molecule moiety, a drug moiety (or payload moiety), and a linker to join the drug moiety to the antibody. In some embodiments the antibody-drug conjugate consists of an antibody moiety, a drug moiety (or payload moiety), and a linker to join the drug moiety to the antibody.

[0452] The antigen binding molecule moiety may be a molecule that binds to a given target antigen. Antigenbinding molecules include antibodies {i.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. The linker may be cleavable or non-cleavable. The linker may be based on a chemical motifs such as disulfides, hydrazones or peptides (cleavable), or thioethers (non-cleavable). The type of linker, cleavable or noncleavable, lends specific properties to the cytotoxic drug. For example, a non-cleavable linker keeps the drug within the cell. As a result, the entire antibody, linker and cytotoxic (anti-cancer) agent enter the targeted cancer cell where the antibody is degraded into an amino acid. The resulting complex - amino acid, linker and cytotoxic agent - is considered to be the active drug. In contrast, cleavable linkers are detached by enzymes in the cancer cell.

[0453] The drug moiety (or payload) may be a small molecule or a nucleic acid drug. In some embodiments, the drug moiety (or payload), is or comprises a cytotoxic agent. In some embodiments, the drug moiety is or comprises a chemotherapeutic agent. In some embodiments, the drug moiety is or comprises an antiarthritis drug. In some embodiments, the drug moiety is or comprises a steroid.

[0454] Functional properties of the antigen-binding molecules

[0455] The antigen-binding molecules described herein may be characterised by reference to certain functional properties. In some embodiments, the antigen-binding molecule described herein may possess one or more of the following properties: binds to LRG1 (e.g. human LRG1 , mouse LRG1 , and / or cynomolgus monkey LRG1 ); binds cross-reactively to human LRG1 and an orthologue (e.g. mouse LRG1 and / or cynomolgus monkey LRG1 ) binds to the leucine-rich C-terminal (LRRCT) of LRG1 ; reduces a function of LRG1 ; reduces or inhibits fibrotic gene expression; reduces or inhibits ECM protein expression (e.g. fibronectin expression); reduces or inhibits migration of cells (e.g. RPE cells); reduces or inhibits proliferation of cells (e.g. RPE cells); reduces or inhibits transdifferentiation of cells (e.g. RPE cells); reduces or inhibits fibroblast activation; reduces or inhibits angiogenesis; reduces or inhibits fibrosis; reduces the pathology of a disease / condition characterised by angiogenesis in a subject; reduces the pathology of a disease / condition characterised by fibrosis in a subject; reduces or inhibits tumour growth; reduces or inhibits metastasis; reduces or inhibits inflammation.

[0456] It will be appreciated that a given antigen-binding molecule may display more than one of the properties recited in the preceding paragraph. A given antigen-binding molecule 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.

[0457] 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 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 {e.g. a dilution series). It will be appreciated that the cells preferably express the target antigen for the antigen-binding molecule {i.e.

[0458] LRG1 ).

[0459] 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 LRG1 may be the concentration of the antigen-binding molecule at which 50% of the maximal level of binding to human LRG1 is achieved.

[0460] 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.

[0461] The antigen-binding molecules described herein bind to LRG1 . The antigen-binding molecules and antigen-binding domains described herein preferably display specific binding to the relevant target antigen {e.g. LRG1 ). As used herein, ‘specific binding’ refers to binding which is selective for the antigen, and which can be discriminated from non-specific 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.

[0462] 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) 907:41 1 -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.

[0463] 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 (i.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.

[0464] 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.

[0465] In some embodiments, the antigen-binding molecule described herein binds to LRG1 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 LRG1 with sub-micromolar affinity, i.e. KD < 1 x 10-6M. In some embodiments, the antigen-binding molecule described herein binds to LRG1 with an affinity in the nanomolar range, i.e. KD = 9.9 x 107to 1 x 10-9M. In some embodiments, the antigen-binding molecule described herein binds to LRG1 with sub-nanomolar affinity, i.e. KD < 1 x 10-9M. In some embodiments, the antigen-binding molecule described herein binds to LRG1 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 LRG1 with sub-picomolar affinity, i.e. KD < 1 x 10-12M.

[0466] In some embodiments, the antigen-binding molecule described herein binds to human LRG1 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 1 1 herein). In some embodiments, the antigen-binding molecule described herein binds to human LRG1 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 1 1 herein).

[0467] In some embodiments, the antigen-binding molecule described herein binds to human LRG1 with an EC50 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 6 herein).

[0468] In some embodiments, the antigen-binding molecule is cross-reactive for human LRG1 and a homologue thereof (e.g. mouse LRG1 ). As used herein, a ‘cross-reactive’ antigen-binding molecule / domain binds to the target antigens for which the antigen-binding molecule / domain is cross-reactive. For example, an antigen-binding molecule / domain / polypeptide which is cross-reactive for human LRG1 and mouse LRG1 binds to human LRG1 , and is also capable of binding to mouse LRG1 . Similarly, an antigen-binding molecule / domain / polypeptide which is cross-reactive for human LRG1 and monkey (e.g. cynomolgus monkey) LRG1 binds to human LRG1 , and is also capable of binding to monkey (e.g. cynomolgus monkey) LRG1 . Cross-reactive antigen-binding molecules / domains / polypeptides may display specific binding to each of the target antigens.

[0469] In some embodiments, the antigen-binding molecule binds to human LRG1 and mouse LRG1 . In some embodiments, the antigen-binding molecule binds to human LRG1 and monkey (e.g. cynomolgus monkey) LRG1 . In some embodiments, the antigen-binding molecule binds to mouse LRG1 and monkey (e.g. cynomolgus monkey) LRG1 . In some embodiments, the antigen-binding molecule binds to human LRG1 , mouse LRG1 and monkey (e.g. cynomolgus monkey) LRG1 . In some embodiments, the antigenbinding molecule binds to human LRG1 , mouse LRG1 , porcine LRG1 and monkey (e.g. cynomolgus monkey) LRG1 .

[0470] The antigen-binding molecules of the present disclosure may bind to a particular region of interest of LRG1 . Antigen-binding molecules according to the present disclosure may bind to linear epitope of LRG1 , consisting of a contiguous sequence of amino acids (i.e. an amino acid primary sequence). In some embodiments, an antigen-binding molecules may bind to a conformational epitope of LRG1 , consisting of a discontinuous sequence of amino acids of the amino acid sequence.

[0471] 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.

[0472] In some embodiments, the antigen-binding molecule of the present disclosure binds to a domain of LRG1 described herein, e.g. an LRR or the LRRCT domain.

[0473] In some embodiments, the antigen-binding molecule of the present disclosure binds to the LRRCT domain of LRG1 . In some embodiments, the antigen-binding molecule binds to the region of LRG1 shown in SEQ ID NO:183. In some embodiments, the antigen-binding molecule contacts the region of LRG1 shown in SEQ ID NO:183. In some embodiments, the antigen-binding molecule binds to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO:183. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:183.

[0474] As used herein, where an antigen-binding molecule ‘contacts’ a given region, it refers to an antigenbinding molecule binding to the given region (e.g. the LRRCT region) via contact with one or more amino acids of the given region. In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region with an affinity in the micromolar range, i.e. KD = 9.9 x 10-4to 1 x 10-6M. In some embodiments, the antigenbinding molecule described herein binds to the LRRCT region with sub-micromolar affinity, i.e. KD < 1 x 10-6M. In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region 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 the LRRCT region with sub-nanomolar affinity, i.e. KD < 1 x 10-9M. In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region 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 the LRRCT region with sub-picomolar affinity, i.e. KD < 1 x 1012M.

[0475] In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region 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. In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region 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.

[0476] In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region with an EC50 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 6 herein). In some embodiments, the antigen-binding molecule described herein binds to the LRRCT region with an EC50 of 10 nM or less, preferably one of <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.

[0477] In some embodiments, the antigen-binding molecule binds to the region of LRG1 shown in SEQ ID NO:190. In some embodiments, the antigen-binding molecule contacts the region of LRG1 shown in SEQ ID NO:190. In some embodiments, the antigen-binding molecule binds to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO:190. In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:190. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NQ:190.

[0478] In some embodiments, the antigen-binding molecule binds to the region of LRG1 shown in SEQ ID NO:191 . In some embodiments, the antigen-binding molecule contacts the region of LRG1 shown in SEQ ID NO:191 . In some embodiments, the antigen-binding molecule binds to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO:191 . In some embodiments, the epitope of the antigen-binding molecule comprises or consists of the amino acid sequence shown in SEQ ID NO:191 . In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:191 .

[0479] In some embodiments, the antigen-binding molecule does not bind to the region of LRG1 shown in SEQ ID NO:192. In some embodiments, the antigen-binding molecule does not contact the region of LRG1 shown in SEQ ID NO:192. In some embodiments, the antigen-binding molecule does not bind to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO:192. In some embodiments, the antigen-binding molecule does not bind to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:192.

[0480] In some embodiments, an antigen-binding molecule does not target / bind to the LRR region of LRG1 . As used herein, ‘LRR region’ refers to an amino acid sequence having at least 70% (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:195. The LRR region comprises 8 leucine-rich repeats (LRRs) (SEQ ID NO:175, 176, 177, 178, 179, 180, 181 , and 182). In some embodiments, the antigen-binding molecule does not bind to the region of LRG1 shown in SEQ ID NO:175, 176, 177, 178, 179, 180, 181 or 182.

[0481] In some embodiments, the antigen-binding molecule does not bind to the region of LRG1 shown in SEQ ID NO:195. In some embodiments, the antigen-binding molecule does not contact the region of LRG1 shown in SEQ ID NO:195. In some embodiments, the antigen-binding molecule does not bind to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO:195. In some embodiments, the antigen-binding molecule does not bind to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:195.

[0482] In some embodiments, the antigen-binding molecule does not bind to the region of LRG1 shown in SEQ ID NO:196. In some embodiments, the antigen-binding molecule does not contact the region of LRG1 shown in SEQ ID NO:196. In some embodiments, the antigen-binding molecule does not bind to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO:196. In some embodiments, the antigen-binding molecule does not bind to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO:196.

[0483] In some embodiments, the antigen-binding molecule does not bind to the region of LRG1 shown in SEQ ID NO: 175, 176, 177, 178, 179, 180, 181 or 182. In some embodiments, the antigen-binding molecule does not contact the region of LRG1 shown in SEQ ID NO: 175, 176, 177, 178, 179, 180, 181 or 182. In some embodiments, the antigen-binding molecule does not bind to LRG1 via contact with one or more amino acids of the region shown in SEQ ID NO: 175, 176, 177, 178, 179, 180, 181 or 182. In some embodiments, the antigen-binding molecule does not bind to a polypeptide comprising or consisting of the amino acid sequence shown in SEQ ID NO: 175, 176, 177, 178, 179, 180, 181 or 182. 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.

[0484] In some embodiments, the antigen-binding molecule is capable of binding the same region of LRG1 , or an overlapping region of LRG1 , to the region of LRG1 which is bound by an antibody comprising the VH and VL regions (see e.g. Table C) of one of clones EBC58, EBC59, EBC60, EBC61 , EBC1191 , EBC1192, EBC1193, EBC1194, EBC1195, EBC1196, EBC1197, EBC1198, EBC1199, EBC1200, EBC1201 , EBC1202, EBC1203, EBC1204, EBC1205, EBC1206, EBC1207, EBC1208, EBC1209, EBC1210, EBC1211 and EBC1212.

[0485] 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.

[0486] In some embodiments, the antigen-binding molecule reduces / inhibits a function of LRG1 .

[0487] In some embodiments, the antigen-binding molecule is an antagonist of LRG1 . In some embodiments, the antigen-binding molecule is capable of inhibiting a function or process mediated by LRG1 . Herein, ‘inhibition’ refers to a reduction, decrease or lessening relative to a control condition. Suitable assays for investigating the function of LRG1 are well known to the skilled person.

[0488] Assays for the identification of antigen-binding molecules capable of reducing / inhibiting a function of LRG1 may comprise treating cells / tissue in the presence of LRG1 with a test antigen-binding molecule, and subsequently comparing the level of relevant function to the level observed in an appropriate control condition {e.g. untreated or control treated cells / tissues).

[0489] Antigen-binding molecules capable of reducing / inhibiting a function of LRG1 , may be identified using assays comprising detecting a correlate of a function of LRG1 {e.g. the gene and / or protein expression, and / or activity, or one or more proteins whose expression is directly / indirectly upregulated or downregulated as a consequence of a function of LRG1 . Such assays may comprise treating cells / tissue in the presence of LRG1 with the test 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 LRG1 to be observed) comparing the level of the correlate of a function of LRG1 in such cells / tissue to the level of the correlate of the relevant function in an appropriate control condition {e.g. untreated or control treated cells / tissues).

[0490] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting a function of LRG1 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.

[0491] A correlate of a function of LRG1 may be e.g. fibrotic gene expression, ECM protein expression, cell migration, cell proliferation, epithelial-mesenchymal transition (EMT) {e.g. by perivascular cells or retinal epithelial cells), fibroblast generation, myofibroblast generation, fibroblast activation, angiogenesis, fibrosis, inflammation, tumorigenesis, ALK1 / Smad1 / 5 / 8 signalling, ALK5 / Smad2 / 3 signalling, BMP signalling, TNFa, or EGF signalling.

[0492] In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not requiring or involving Fc-mediated function. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 independently of Fc-mediated function. That is, in some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 in an Fc region-independent manner.

[0493] The ability of an antigen-binding molecule to inhibit a function of LRG1 by a mechanism not requiring / involving Fc-mediated function can be evaluated e.g. by analyzing the ability of the antigenbinding molecule provided in a format lacking a functional Fc region to inhibit a function of LRG1 . For example, the effect on a function of LRG1 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.).

[0494] In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not involving ADCC. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not involving ADCP. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not involving CDC.

[0495] In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 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 LRG1 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 LRG1 by a mechanism not requiring binding of the antigen-binding molecule to one or more of FcyRI, FcyRlla, FcyRllb, FcyRllc, FcyRllla and FcyRlllb. In some embodiments, the antigenbinding molecule is able to inhibit a function of LRG1 by a mechanism not requiring binding to FcyRllla. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not requiring binding to FcyRlla. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not requiring binding to FcyRllb. In some embodiments, the antigenbinding molecule is able to inhibit a function of LRG1 by a mechanism not requiring binding to a complement protein. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not requiring binding to C1q. In some embodiments, the antigen-binding molecule is able to inhibit a function of LRG1 by a mechanism not requiring N297 glycosylation.

[0496] In some embodiments, the antigen-binding molecule reduces / inhibits fibrotic gene expression in cells (e.g. in RPE cells, dermal fibroblasts, synovial fibroblasts, mesangial cells). In some embodiments, the antigen-binding molecule reduces / inhibits extracellular matrix (ECM) protein expression in cells (e.g. in RPE cells, dermal fibroblasts, synovial fibroblasts, mesangial cells). In some embodiments, the antigenbinding molecule reduces / inhibits migration of cells (e.g. in RPE cells, dermal fibroblasts, synovial fibroblasts, mesangial cells). In some embodiments, the antigen-binding molecule reduces / inhibits proliferation of cells (e.g. in RPE cells, dermal fibroblasts, synovial fibroblasts, mesangial cells). In some embodiments, the antigen-binding molecule reduces / inhibits transdifferentiation of RPE cells (e.g. differentiation into fibroblasts or myofibroblasts). In some embodiments, the antigen-binding molecule reduces / inhibits fibroblast activation.

[0497] The ability of an antigen-binding molecule to reduce / inhibit fibrotic gene expression / ECM protein expression / migration of cells / proliferation of cells / transdifferentiation of RPE cells / fibroblast activation can be determined for example by analysis of fibrotic gene expression / ECM protein expression / migration of cells / proliferation of cells / transdifferentiation of RPE cells / fibroblast activation in the presence of, or following incubation with, the antigen-binding molecule. An antigen-binding molecule which is capable of inhibiting fibrotic gene expression / ECM protein expression / migration of cells / proliferation of cells / transdifferentiation of RPE cells / fibroblast activation is identified by the observation of a reduction / decrease in the level of fibrotic gene expression / ECM protein expression / migration of cells / proliferation of cells / transdifferentiation of RPE cells / fibroblast activation in the presence of the antigen-binding molecule, as compared to the level of fibrotic gene expression / ECM protein expression / migration of cells / proliferation of cells / transdifferentiation of RPE cells / fibroblast activation in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule).

[0498] Antigen-binding molecules capable of reducing / inhibiting fibrotic gene expression may be identified using assays comprising detecting the level of fibrotic gene expression. Gene expression can be measured by various means known to those skilled in the art, for example by measuring levels of mRNA by quantitative real-time PCR (qRT-PCR), or by reporter-based methods. In preferred embodiments, fibrotic gene expression can be evaluated in an assay essentially as described in Example 7.

[0499] As used herein, a ‘fibrotic gene’ is a gene associated with fibrosis or fibrotic disease, e.g. expression of the gene is associated with fibrosis or fibrotic disease. Examples of genes associated with fibrosis / fibrotic disease include CTGF, Coll a1 , Col4a2, fibronectin, SMAD3, TGFB1 , LOX, LOXL2, PPARG, TIMP1 , TIMP2, NGF, CD44, ICAM1 , MMP12 and HSPB1 . Genes associated with fibrosis are reviewed in Gu et al (2021 ) Front. Genet. 11 :627396, which is hereby incorporated by reference in its entirety.

[0500] Antigen-binding molecules capable of reducing / inhibiting ECM protein expression may be identified using assays comprising detecting the level of ECM protein expression. Protein expression can be measured by various methods well known in the art, e.g. by antibody-based methods, for example by western blot, immunohistochemistry, immunocytochemistry, flow cytometry, ELISA, ELISPOT, or reporter-based methods. In preferred embodiments, ECM protein expression can be evaluated in an assay essentially described in Example 7.

[0501] As used herein, an ‘ECM protein’ is a protein that is a component of / forms part of the extracellular matrix (ECM). Examples of proteins found in the ECM include fibronectin, collagen, laminin and elastin.

[0502] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting fibrotic gene expression / ECM protein expression 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 fibrotic gene expression / ECM protein expression observed in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), in a given assay.

[0503] Antigen-binding molecules capable of reducing / inhibiting migration of cells {e.g. RPE cells) may be identified using assays comprising detecting the level of cell migration. Cell migration can be measured by various method well known in the art, e.g. in a transwell assay. In preferred embodiments, cell migration can be evaluated in an assay essentially as described in Example 7.

[0504] Antigen-binding molecules capable of reducing / inhibiting proliferation of cells {e.g. RPE cells) may be identified using assays comprising detecting the level of cell proliferation. Cell proliferation can be measured by various methods well known in the art, e.g. detecting a marker of cell proliferation {e.g. Ki67),3H thymidine incorporation or CFSE dilution assays. In preferred embodiments, cell proliferation can be evaluated in an assay essentially as described in Example 8.

[0505] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting migration of cells / prol iteration of cells 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 migration of cells / proliferation of cells observed in the absence of the antigen-binding molecule (or in the presence of an appropriate control antigen-binding molecule), in a given assay. Antigen-binding molecules capable of reducing / inhibiting transdifferentiation of cells may be identified using an assay comprising detecting the level of transdifferentiation of cells or the level of a correlate of transdifferentiation of cells.

[0506] As used herein, ‘transdifferentiation’ refers to the conversion of one cell type to another. It may be a process in which one mature somatic cell is transformed into another mature somatic cell without undergoing an intermediate pluripotent state or progenitor cell type. In some embodiments, an epithelial cell {e.g. a retinal pigment epithelial cell) may undergo transdifferentiation. In some embodiments, a pericyte {e.g. a retinal vascular pericyte) may undergo transdifferentiation. In some embodiments, a given cell may undergo transdifferentiation into a mesenchymal cell, a fibroblast or a myofibroblast. In some embodiments transdifferentiation may be from an RPE cell to a fibroblast. In some embodiments transdifferentiation may be from an RPE cell to a myofibroblast. In some embodiments transdifferentiation may be from an RPE cell to a mesenchymal cell {i.e. epithelial-mesenchymal transition (EMT)).

[0507] Transdifferentiation of a given cell type can be measured by detecting a reduction in the amount / proportion / activity of the original cell type and / or by an increase in the amount / proportion / activity of the transdifferentiated cell type. Characteristics of a given cell type and cell markers for a given cell type are well known to the skilled person.

[0508] For example, RPE cell markers include MITF, 0TX2, RPE65, LRAT, CRALBP, PEDF, BEST1 and ZO1 . The molecular signature of RPE cells is reviewed in Liao et al. Hum. Mol. Genet (2010) 19(21 ): 4229- 4238, which is incorporated by reference herein. Fibroblasts may be characterised by gene or protein expression of one or more of COL1 A, ACTA2, prolyl-4-hydroxylase, MAS516, and FSP1 . Identification of fibroblasts is reviewed in Lendahl et al. Nat Commun (2022) 13:3409. Myofibroblast markers may include one or more of aSMA, vimentin, palladin, cofilin or desmin, and p-cadherins. Myofibroblast markers are reviewed in Younesi et al. Methods in Molecular Biology (2021 ) Myofi brobasts, pages 17-47, which is incorporated by reference herein. Gene expression can be measured by various means known to those skilled in the art, for example by measuring levels of mRNA by quantitative real-time PCR (qRT-PCR), or by reporter-based methods. Similarly, protein expression can be measured by various methods well known in the art, e.g. by antibody-based methods, for example by western blot, immunohistochemistry, immunocytochemistry, flow cytometry, ELISA, ELISPOT, or reporter-based methods.

[0509] In preferred embodiments, transdifferentiation of RPE cells can be evaluated in an assay essentially as described in Example 9.

[0510] Antigen-binding molecules capable of reducing / inhibiting fibroblast activation may be identified using an assay comprising detecting the level of fibroblast activation. Fibroblast activation refers to the transition of the cell from a quiescent state to an activated state. Activated fibroblasts may exhibit one or more of increased proliferation, increased migration and synthesis of ECM proteins. Fibroblasts according to the present disclosure may be derived from any tissue, including eye, liver, lungs, kidney, heart, blood vessels, skin, pancreas, spleen, bowel {e.g. large or small intestine), brain, and bone marrow. In particular embodiments, for the purposes of analysis of the antigen-binding molecule, the fibroblasts may be eye or bone marrow fibroblasts. Fibroblasts may be characterised by gene or protein expression of one or more of COL1 A, ACTA2, prolyl-4-hydroxylase, MAS516, and FSP1 .

[0511] In preferred embodiments, fibroblast activation can be evaluated in an assay essentially as described in Example 7 or Example 8.

[0512] In some embodiments, an antigen-binding molecule is capable of reducing / inhibiting angiogenesis in vitro or ex vivo. In some embodiments, an antigen-binding molecule is capable of reducing / inhibiting angiogenesis {e.g. ocular angiogenesis, e.g. choroidal neovascularization) in a subject. In some embodiments, an antigen-binding molecule is capable of reversing angiogenesis in vitro or ex vivo. In some embodiments, an antigen-binding molecule is capable of reversing angiogenesis {e.g. ocular angiogenesis, e.g. choroidal neovascularization) in a subject.

[0513] Inhibition of angiogenesis can be measured in vitro, ex vivo or in vivo. For example, whether an antigenbinding molecule is capable of inhibiting angiogenesis in a given tissue can be analysed by treating cells derived from that tissue with a proangiogenic stimulus, and then analysing whether the antigen-binding molecule can reduce angiogenesis or a marker of angiogenesis. Whether an antigen-binding molecule is capable of inhibiting angiogenesis in a given tissue can be analysed ex vitro by treating an explant from that tissue with a proangiogenic stimulus {e.g. LRG1 ), and then analysing whether the antigen-binding molecule can reduce angiogenesis or a marker of angiogenesis. Whether an antigen-binding molecule is capable of inhibiting angiogenesis can be analysed in vivo, for example, by administering the antigenbinding molecule to a subject {e.g. a subject that has been exposed to a proangiogenic stimulus), and analysing tissue(s) for angiogenesis or one or more markers of angiogenesis.

[0514] Angiogenesis may be measured by means well known to the skilled person, for example by analysing gene or protein expression of one of more markers of angiogenesis {e.g. VEGF), by immunofluorescence staining of cell surface markers of blood vessels (e.g. CD31 ), or by angiography {e.g. fundus fluorescein angiography).

[0515] In preferred embodiments, angiogenesis can be evaluated in an assay essentially as described in Example 9, 10 or 12.

[0516] In some embodiments, an antigen-binding molecule is capable of reducing / inhibiting fibrosis in vitro or ex vivo. In some embodiments, an antigen-binding molecule is capable of reducing / inhibiting fibrosis {e.g. ocular fibrosis, e.g. subretinal fibrosis) in a subject. In some embodiments, an antigen-binding molecule is capable of reversing fibrosis in vitro or ex vivo. In some embodiments, an antigen-binding molecule is capable of reversing fibrosis {e.g. ocular fibrosis, e.g. subretinal fibrosis) in a subject. Inhibition of fibrosis can be measured in vitro, ex vivo or in vivo. For example, whether an antigen-binding molecule is capable of inhibiting fibrosis in a given tissue can be analysed in vitro by treating cells derived from that tissue with a profibrotic stimulus (e.g. TGFp or LRG1), and then analysing whether the antigenbinding molecule can reduce a marker of fibrosis. Whether an antigen-binding molecule is capable of inhibiting fibrosis in a given tissue can be analysed ex vivo by treating an explant from that tissue with a profibrotic stimulus (e.g. TGFp or LRG1), and then analysing whether the antigen-binding molecule can reduce a marker of fibrosis. Whether an antigen-binding molecule is capable of inhibiting fibrosis can be analysed in vivo, for example, by administering the antigen-binding molecule to a subject (e.g. a subject that has been exposed to a profibrotic stimulus), and analysing tissue(s) for one or more markers of fibrosis.

[0517] Fibrosis may be measured by means well known to the skilled person, for example by analysing gene or protein expression of one or more markers of fibrosis in a given tissue or tissues. Markers of fibrosis include increased level of collagen, fibronectin, CTGF, aSMA, periostin, IL-6, IL-11 , TIMP1 and MMP2, extracellular matrix components.

[0518] In preferred embodiments, fibrosis can be evaluated in an assay essentially as described in Example 9, 10 or 12.

[0519] In some embodiments, the antigen-binding molecule is capable of reducing / inhibiting fibrosis and angiogenesis in vitro or ex vivo. In some embodiments, the antigen-binding molecule is capable of reducing / inhibiting fibrosis and angiogenesis in a subject.

[0520] In some embodiments, the antigen-binding molecule reduces / inhibits the pathology of a disease / condition characterised by fibrosis in a subject. In some embodiments, the antigen-binding molecule reduces / inhibits the pathology of a disease / condition characterised by angiogenesis in a subject. In some embodiments, the antigen-binding molecule reduces / inhibits the pathology of a disease / condition characterised by fibrosis and angiogenesis in a subject. In some embodiments the antigen-binding molecule reduces / inhibits the pathology of macular degeneration. In some embodiments the antigenbinding molecule reduces / inhibits the pathology of rheumatoid arthritis. In some embodiments, the antigen-binding molecule reduces / inhibits the pathology of a cancer. In some embodiments, the antigenbinding molecule reduces / inhibits the pathology of diabetic nephropathy. In some embodiments, the antigen-binding molecule reduces / inhibits the pathology of inflammatory bowel disease.

[0521] In some embodiments, the antigen-binding molecule of the present disclosure is capable of reducing / inhibiting pathology of a disease / condition characterised by fibrosis and / or angiogenesis in a subject 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 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 the pathology of the disease / condition), in a given assay.

[0522] Chimeric antigen receptors (CARs)

[0523] The present disclosure also provides Chimeric Antigen Receptors (CARs) comprising the antigen-binding polypeptides or polypeptides of the present disclosure.

[0524] CARs are recombinant receptors that provide both antigen-binding and T cell activating functions. CAR structure and engineering is reviewed, for example, in Dotti et al., Immunol Rev (2014) 257(1 ), hereby incorporated by reference in its entirety. CARs comprise an antigen-binding region linked to a cell membrane anchor region and a signalling region. An optional hinge region may provide separation between the antigen-binding region and cell membrane anchor region, and may act as a flexible linker.

[0525] The CAR of the present disclosure comprises an antigen-binding region which comprises or consists of the antigen-binding molecule of the present disclosure, or which comprises or consists of a polypeptide according to the present disclosure.

[0526] The cell membrane anchor region is provided between the antigen-binding region and the signalling region of the CAR and provides for anchoring the CAR to the cell membrane of a cell expressing a CAR, with the antigen-binding region in the extracellular space, and signalling region inside the cell. In some embodiments, the CAR comprises a cell membrane anchor region comprising or consisting of an amino acid sequence which comprises, consists of, or is derived from, the transmembrane region amino acid sequence for one of CD3- , CD4, CD8 or CD28. As used herein, a region which is ‘derived from’ a reference amino acid sequence comprises an amino acid sequence having at least 60% (e.g. one of >60%, >65%, >70%, >75%, >80%, >85%, >86%, >87%, >88%, >89%, >90%, >91 %, >92%, >93%, >94%, >95%, >96%, >97%, >98%, >99% or 100%) sequence identity to the reference sequence.

[0527] The signalling region of a CAR allows for activation of the T cell. The CAR signalling regions may comprise the amino acid sequence of the intracellular domain of CD3- , which provides immunoreceptor tyrosine-based activation motifs (ITAMs) for phosphorylation and activation of the CAR-expressing T cell. Signalling regions comprising sequences of other ITAM-containing proteins such as FcyRI have also been employed in CARs (Haynes et al., 2001 J Immunol 166(1 ):182-187). Signalling regions of CARs may also comprise co-stimulatory sequences derived from the signalling region of co-stimulatory molecules, to facilitate activation of CAR-expressing T cells upon binding to the target protein. Suitable co-stimulatory molecules include CD28, 0X40, 4-1 BB, ICOS and CD27. In some cases CARs are engineered to provide for co-stimulation of different intracellular signalling pathways. For example, signalling associated with CD28 costimulation preferentially activates the phosphatidylinositol 3-kinase (PI3K) pathway, whereas the 4-1 BB-mediated signalling is through TNF receptor associated factor (TRAF) adaptor proteins. Signalling regions of CARs therefore sometimes contain co-stimulatory sequences derived from signalling regions of more than one co-stimulatory molecule. In some embodiments, the CAR of the present disclosure comprises one or more co-stimulatory sequences comprising or consisting of an amino acid sequence which comprises, consists of, or is derived from, the amino acid sequence of the intracellular domain of one or more of CD28, 0X40, 4-1 BB, ICOS and CD27.

[0528] An optional hinge region may provide separation between the antigen-binding domain and the transmembrane domain, and may act as a flexible linker. Hinge regions may be derived from IgG 1 . In some embodiments, the CAR of the present disclosure comprises a hinge region comprising or consisting of an amino acid sequence which comprises, consists of, or is derived from, the amino acid sequence of the hinge region of IgG 1 .

[0529] Also provided is a cell comprising a CAR according to the present disclosure. The CAR according to the present disclosure may be used to generate CAR-expressing immune cells, e.g. CAR-T or CAR-NK cells. Engineering of CARs into immune cells may be performed during culture, in vitro.

[0530] The antigen-binding region of the CAR of the present disclosure may be provided with any suitable format, e.g. scFv, scFab, etc.

[0531] Nucleic acids and vectors

[0532] The present disclosure provides a nucleic acid, or a plurality of nucleic acids, encoding an antigen-binding molecule, polypeptide or CAR according to the present disclosure. In some embodiments, the nucleic acid(s) comprise or consist of DNA and / or RNA.

[0533] 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, polypeptide or CAR according to the present disclosure may be produced within a cell by transcription from a vector encoding the antigen-binding molecule, polypeptide or CAR, and subsequent translation of the transcribed RNA.

[0534] 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, polypeptide or CAR according to the present disclosure. The vector may be an expression vector comprising elements required for expressing nucleic acid(s) comprising / encoding an antigen-binding molecule, polypeptide or CAR according to the present disclosure.

[0535] 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.

[0536] The nucleotide sequence may be contained in a vector, e.g. an expression vector. 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.

[0537] 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).

[0538] Suitable vectors include plasmids, binary vectors, DNA vectors, mRNA vectors, 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 or Morgan and Boyerinas, Biomedicines (2016) 4:9, which are both hereby incorporated by reference in their entirety.

[0539] 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.

[0540] Constituent polypeptides of an antigen-binding molecule 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.

[0541] Cells comprisinq / expressinq the antigen-binding molecules and polypeptides

[0542] The present disclosure also provides a cell comprising or expressing an antigen-binding molecule, polypeptide or CAR 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.

[0543] 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). 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, NSO and BHK cells. In preferred embodiments, the cell is, or is derived from, a CHO cell.

[0544] 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).

[0545] The present disclosure also provides a method for producing a cell expressing / comprising an antigenbinding molecule, polypeptide or CAR 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.

[0546] The present disclosure also provides cells obtained or obtainable by the methods according to the present disclosure.

[0547] Producing the antigen-binding molecules and polypeptides

[0548] Antigen-binding molecules and polypeptides according to the present disclosure may be prepared according to methods for the production of polypeptides known to the skilled person.

[0549] Polypeptides 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.

[0550] Alternatively, antigen-binding molecules and 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 molecules 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.

[0551] In some cases, the antigen-binding molecules of the present disclosure are comprised of more than one polypeptide chain. In such cases, production of the antigen-binding molecules may comprise transcription and translation of more than one polypeptide, and subsequent association of the polypeptide chains to form the antigen-binding molecule.

[0552] 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 coli. 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.

[0553] 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.

[0554] 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.

[0555] Production 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).

[0556] 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.

[0557] Following culturing the cells that express the antigen-binding molecule / 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. 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.

[0558] 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.

[0559] 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.

[0560] Compositions

[0561] The present disclosure also provides compositions comprising the antigen-binding molecules, polypeptides, CARs, nucleic acids, expression vectors and cells described herein.

[0562] The antigen-binding molecules, polypeptides, CARs, nucleic acids, expression vectors and cells described herein may be formulated as pharmaceutical compositions or medicaments for clinical use and may comprise a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0563] The compositions 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).

[0564] 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), 23rd Edition (2020), Academic Press.

[0565] Compositions 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.

[0566] Suitable formulations may comprise the relevant article 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.

[0567] In some embodiments, the composition is formulated for injection or infusion, e.g. into a blood vessel, tissue / organ of interest, or tumor.

[0568] 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, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof) or cell described herein; isolating an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof) or cell described herein; and / or mixing an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof) or cell described herein with a pharmaceutically acceptable carrier, adjuvant, excipient or diluent.

[0569] 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 cancer), the method comprising formulating a pharmaceutical composition or medicament by mixing an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof) or cell described herein with a pharmaceutically acceptable carrier, adjuvant, excipient or diluent.

[0570] Therapeutic and prophylactic applications

[0571] The antigen-binding molecules, polypeptides, CARs, nucleic acids, expression vectors, cells and compositions described herein find use in therapeutic and prophylactic methods. The present disclosure provides an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell or composition described herein for use in a method of medical treatment or prophylaxis. Also provided is an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell 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 antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell 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, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell or composition described herein.

[0572] 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 the 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 the development of a later stage of the disease / condition.

[0573] It will be appreciated that the articles 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 LRG1 .

[0574] For example, the disease / condition may be a disease / condition in which LRG1 is pathologically- implicated, e.g. a disease / condition in which an increased level / activity of LRG1 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 LRG1 may be a risk factor for the onset, development or progression of the disease / condition.

[0575] In some embodiments, the disease / condition to be treated in accordance with the present disclosure is a disease / condition characterised by an increase in the level of expression or activity of LRG1 , e.g. as compared to the level of expression / activity in the absence of the disease / condition.

[0576] Treatment in accordance with the methods of the present disclosure may achieve a reduction in the level of expression / activity of LRG1 in a subject (compared to an equivalent untreated subject, or a subject treated with an appropriate control).

[0577] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by elevated LRG1 expression / activity. Diseases / conditions associated with LRG1 dysfunction are reviewed in Camilli et al. Journal of Biomedical Science (2022) 29(1 ):6, which is incorporated by reference herein.

[0578] Antigen-binding molecules according to the present invention are useful in the treatment of diseases characterised by upregulation in LRG1 expression / activity.

[0579] Diseases characterised by upregulation in LRG1 expression include: cancer such as glioblastoma, retinoblastoma, oral squamous cell carcinoma, esophageal squamous cell carcinoma, non small cell lung cancer, squamous cell lung carcinoma, breast cancer, clear cell renal cell carcinoma, hepatocellular carcinoma, gastric cancer, pancreatic cancer, biliary tract carcinoma, colorectal carcinoma, ovarian cancer, cervical cancer, endometrial cancer, bladder cancer, prostate cancer, leukaemia; Alzheimer’s disease, Parkinson’s disease, progressive supranuclear palsy, stroke, hydrocephalus, neovascular AMD, diabetic retinopathy, corneal neovascularisation, idiopathic pulmonary fibrosis, asthma, bronchopulmonary dysplasia, SARS-CoV-2 acute respiratory distress syndrome, emphysema, myocardial infarction, atherosclerosis, idiopathic pulmonary hypertension, diabetic kidney disease, idiopathic nephrotic syndrome, lupus nephritis, vasculitides, colitis, rheumatoid arthritis, osteoarthritis, Still’s disease, viral and bacterial infection, diabetes, acute pancreatitis and psoriasis.

[0580] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a fibroinflammatory disease / condition.

[0581] As used herein, a ‘fibroinflammatory disease’ or ‘fibroinflammatory condition’ refers to a disease / condition associated with fibroinflammation (i.e. associated with fibrosis and / or inflammation). A fibroinflammatory disease / condition may be associated with fibroinflammatory processes / pathways (i.e. fibrotic and / or inflammatory processes / pathways).

[0582] Fibroinflammatory disorders are reviewed in e.g. Systemic Fibroinflammatory disorders, 2017 (Springer, DOI: 10.1007 / 978-3-319-41349-5), which is hereby incorporated by reference in its entirety.

[0583] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure may be fibroinflammation, a fibroinflammatory condition, or a disease / disorder characterised by fibroinflammation.

[0584] In some embodiments, the disease / disorder may be fibrosis, a fibrotic condition, or a disease / disorder characterised by fibrosis. In some embodiments, the disease / disorder may be inflammation, an inflammatory condition, or a disease / disorder characterised by inflammation. In some embodiments, the disease / disorder may be fibrosis and inflammation, a fibrotic and inflammatory condition, or a disease / disorder characterised by fibrosis and inflammation.

[0585] Inflammatory reactions play an important part in triggering fibrosis in many different organ systems. Inflammation can lead to excess in deposition of ECM components in the affected tissues. Low-grade but persistent inflammation is also thought to contribute to the progression of fibrosis in cardiovascular disease and hypertension. In many fibrotic disorders, a persistent inflammatory trigger is crucial to upregulation of production of growth factors, proteolytic enzymes, angiogenic factors and fibrogenic cytokines, which stimulate the deposition of connective tissue elements that progressively remodel and destroy normal tissue architecture.

[0586] As used herein, “fibrosis” refers to the formation of excess fibrous connective tissue as a result of the excess deposition of extracellular matrix components, for example collagen. Fibrous connective tissue is characterised by having extracellular matrix (ECM) with a high collagen content. The collagen may be provided in strands or fibers, which may be arranged irregularly or aligned. The ECM of fibrous connective tissue may also include glycosaminoglycans.

[0587] As used herein, “excess fibrous connective tissue” refers to an amount of connective tissue at a given location (e.g. a given tissue or organ, or part of a given tissue or organ) which is greater than the amount of connective tissue present at that location in the absence of fibrosis, e.g. under normal, non- pathological conditions. As used herein, “excess deposition of extracellular matrix components” refers to a level of deposition of one or more extracellular matrix components which is greater than the level of deposition in the absence of fibrosis, e.g. under normal, non-pathological conditions.

[0588] The cellular and molecular mechanisms of fibrosis are described in Wynn, J. Pathol. (2008) 214(2): 199- 210, and Wynn and Ramalingam, Nature Medicine (2012) 18:1028-1040, which are hereby incorporated by reference in their entirety.

[0589] Damage to tissues can result from various stimuli, including infections, autoimmune reactions, toxins, radiation and mechanical injury. Repair typically involves the replacement of injured cells by cells of the same type, and replacement of normal parenchymal tissue with connective tissue. Repair processes become pathologic when they are not controlled properly, resulting in excess deposition of ECM components in which normal parenchymal tissue is replaced with connective tissue. In diseases such as idiopathic pulmonary fibrosis, liver cirrhosis, cardiovascular fibrosis, systemic sclerosis and nephritis, extensive tissue remodelling and fibrosis can ultimately lead to organ failure and death.

[0590] The main cellular effectors of fibrosis are myofibroblasts. In response to tissue injury, damaged cells and leukocytes produce pro-fibroinflammafory factors such as TGFp, IL-13 and PDGF, which activate fibroblasts (and other myofibroblast precursor cells) to become aSMA-ex pressing myofibroblasts, and recruit myofibroblasts to the site of injury. Myofibroblasts produce large amounts of extracellular matrix components such as collagen and periostin for wound contracture and closure, and also produce proinflammatory cytokines such as IL-6, and tissue remodelling factors such as MMP2 and TIMP1 . Persistent / chronic infection and / or inflammation can result in the generation of too many myofibroblasts, and consequently the over-production of extracellular matrix, resulting in fibrosis. In many diseases and conditions characterised by fibrosis, a persistent inflammatory trigger is crucial to upregulation of production of growth factors, proteolytic enzymes, angiogenic factors and fibrogenic cytokines, which stimulate the deposition of connective tissue elements that progressively remodel and destroy normal tissue architecture.

[0591] Fibrosis can be triggered by pathological conditions, e.g. conditions, infections or disease states that lead to production of pro-fibrotic factors such as TGFpl . In some embodiments, fibrosis may be caused by physical injury / stimuli, chemical injury / stimuli or environmental injury / stimuli. Physical injury / stimuli may occur during surgery, e.g. iatrogenic causes. Chemical injury / stimuli may include drug induced fibrosis, e.g. following chronic administration of drugs such as bleomycin, cyclophosphamide, amiodarone, procainamide, penicillamine, gold and nitrofurantoin (Daba et al., Saudi Med J 2004 Jun; 25(6): 700-6). Environmental injury / stimuli may include exposure to asbestos fibres or silica.

[0592] Fibrosis can be of any tissue / organ of the body. In some embodiments, fibrosis is of the lung {e.g. bronchioles, alveoli), airways {e.g. nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi), heart, kidney, liver, skeletal muscle, blood vessels, eye, skin, pancreas, bowel, small intestine, large intestine, colon, joints, brain, or bone marrow. Fibrosis may also occur in multiple tissues / organs at once.

[0593] In some embodiments, fibrosis may be of an organ or tissue of the respiratory system, e.g. the lung {e.g. bronchioles, alveoli), or airways {e.g. nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi). In some embodiments, fibrosis may be of an organ or tissue of the cardiovascular system, e.g. the heart or blood vessels. In some embodiments, fibrosis may be of an organ or tissue of the gastrointestinal system, e.g. of the liver, bowel, small intestine, large intestine, colon, or pancreas. In some embodiments, fibrosis may be of the eye. In some embodiments, fibrosis may be of the skin. In some embodiments, fibrosis may be of an organ or tissue of the nervous system, e.g. the brain. In some embodiments, fibrosis may be of the bone marrow. In some embodiments, fibrosis may be of the joints. In some embodiments, fibrosis may be of an organ or tissue of the urinary system, e.g. the kidneys. In some embodiments, fibrosis may be of an organ or tissue of the musculoskeletal system, e.g. muscle tissue. In some embodiments, fibrosis may be of an organ or tissue of one or more organ systems. In some embodiments, fibrosis may be of the eye.

[0594] As used herein, a disease / condition which is ‘characterised by fibrosis’ is a disease / condition in which fibrosis is a symptom of the disease / condition. Diseases and conditions characterised by fibrosis include, but are not limited to:

[0595] Diseases / conditions affecting the respiratory system such as pulmonary fibrosis, fibrothorax, radiation-induced lung injury, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (I PF), cystic fibrosis, progressive massive fibrosis, scleroderma, obliterative bronchiolitis, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor stroma in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis and asthma;

[0596] Diseases / conditions affecting the liver such as chronic liver disease, liver fibrosis, bridging fibrosis, cirrhosis, non-alcoholic fatty liver disease (NAFLD), steatohepatitis, non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcoholic fatty liver (AFL), alcoholic hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomal liver disease and hepatocellular carcinoma (HCC); Diseases / conditions affecting the cardiovascular system such as hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), fibrosis of the atrium, atrial fibrillation, fibrosis of the ventricle, ventricular fibrillation, myocardial fibrosis, interstitial fibrosis, replacement fibrosis Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), atherosclerosis, arterial stiffness, chronic pulmonary hypertension, AIDS-associated pulmonary hypertension, heart failure with preserved ejection fraction (HFpEF), heart failure with reduced ejection fraction (HFrEF), varicose veins and cerebral infarcts;

[0597] Diseases / conditions affecting the kidneys such as tubulointerstitial fibrosis, glomerular fibrosis, renal fibrosis, nephritic syndrome, Alport's syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry's disease, diabetic nephropathy, chronic glomerulonephritis and nephritis associated with systemic lupus;

[0598] Diseases / conditions affecting the pancreas such as pancreatic fibrosis, cystic fibrosis, pancreatitis (e.g. chronic pancreatitis, acute pancreatitis);

[0599] Diseases / conditions affecting the nervous system such as gliosis, Alzheimer's disease and multiple sclerosis;

[0600] Diseases / conditions affecting the musculoskeletal system such as muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker’s muscular dystrophy (BMD) and fibrotic myopathy;

[0601] Diseases / conditions affecting the gastrointestinal system such as inflammatory bowel disease (IBD), Crohn’s disease, ulcerative colitis, microscopic colitis and primary sclerosing cholangitis (PSC);

[0602] Diseases / conditions affecting the skin such as scleroderma, nephrogenic systemic fibrosis, Dupuytren’s contracture, cutis keloid and diabetic wounds;

[0603] Diseases / conditions affecting the eye such as macular degeneration, Age-related Macular Degeneration (AMD), Geographic Atrophy (‘dry’ or non-exudative AMD), early AMD, early onset macular degeneration (EOMD), intermediate AMD, late / advanced AMD, ‘wet’ (neovascular or exudative) AMD, choroidal neovascularisation (CNV), retinal dystrophy, glaucoma (open-angle or closed-angle), neuromyelitis optica (neuromyelitis optica spectrum disorder (NMOSD)), diabetic macular edema (DME), diabetic retinopathy, proliferative diabetic retinopathy (PDR), polypoidal choroidal vasculopathy, proliferative vitreoretinopathy (PVR), macular edema, drusen formation, Grave's ophthalmopathy, corneal opacification, subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis {e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis and preretinal fibrosis;

[0604] Diseases / conditions affecting the joints such as arthrofibrosis, arthritis and adhesive capsulitis;

[0605] Diseases / conditions affecting multiple tissues / organ systems, including progressive systemic sclerosis (PSS), chronic graft versus host disease (GVHD); fibrotic pre-neoplastic and fibrotic neoplastic disease, and fibrosis induced by chemical or environmental insult {e.g., cancer chemotherapy, pesticides, radiation / cancer radiotherapy);

[0606] Cancers, such as hepatocellular carcinoma, gastric cancer, oesophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, and vulvar cancer;

[0607] Mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis and Peyronie’s disease. It will be appreciated that many of the diseases / conditions listed above are interrelated.

[0608] In some embodiments, the fibrosis is fibrosis of the eye ( / '.e. ocular fibrosis). In some embodiments, the fibrosis is selected from: subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis {e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis and preretinal fibrosis.

[0609] In some embodiments, the fibrosis is associated with one or more of: macular degeneration, Age-related Macular Degeneration (AMD), Geographic Atrophy (‘dry’ or non-exudative AMD), early AMD, early onset macular degeneration (EOMD), intermediate AMD, late / advanced AMD, ‘wet’ (neovascular or exudative) AMD, choroidal neovascularisation (CNV), retinal dystrophy, glaucoma (open-angle or closed-angle), neuromyelitis optica (neuromyelitis optica spectrum disorder (NMOSD)), diabetic macular edema (DME), diabetic retinopathy, proliferative diabetic retinopathy (PDR), polypoidal choroidal vasculopathy, proliferative vitreoretinopathy (PVR), macular edema, drusen formation, Grave's ophthalmopathy, corneal opacification, subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis {e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis and preretinal fibrosis.

[0610] As used herein “early AMD” refers to a stage of AMD characterised by the presence of medium-sized drusen, commonly having a width of up to -200 pm, within the Bruch’s membrane adjacent to the RPE layer. Subjects with early AMD typically do not present significant vision loss. As used herein “intermediate AMD” refers to a stage of AMD characterised by large drusen and / or pigment changes in the retina. Intermediate AMD may be accompanied by some vision loss. As used herein “late AMD” refers to a stage of AMD characterised by the presence of drusen and vision loss due to damage to the macula. In all stages of AMD, ‘reticular pseudodrusen’ (RPD) or ‘reticular drusen’ may be present, referring to the accumulation of extracellular material in the subretinal space between the neurosensory retina and RPE. “Late AMD” encompasses ‘dry’ and ‘wet’ AMD. In ‘dry’ AMD (also known as geographic atrophy), there is a gradual breakdown of the light-sensitive cells in the macula that convey visual information to the brain and of the supporting tissue beneath the macula. In ‘wet’ AMD (also known as choroidal neovascularization AMD (nAMD) and exudative AMD), abnormal blood vessels grow underneath and into the retina. These vessels can leak fluid and blood which can lead to swelling and damage of the macula and subsequent fibrosis / scar formation. The damage may be rapid and severe.

[0611] Fibrosis of the eye and fibrotic diseases of the eye are reviewed e.g. in Mallone et al., Int J Mol Sci. (2021 ) 22:11748 and Friedlander, J Clin Invest. (2007) 117(3):576-586 which are hereby incorporated by reference in their entirety. Neovascular AMD (nAMD) is reviewed e.g. in Pugazhendhi et al., Int J Mol Sci. (2021 ) 22(3) :1170, which is hereby incorporated by reference in its entirety.

[0612] In some embodiments, the fibrosis is fibrosis of the pancreas {i.e. pancreatic fibrosis). In some embodiments, the fibrosis is selected from: interlobular fibrosis, periductal fibrosis, diffuse interlobular fibrosis and diffuse intralobular fibrosis. In some embodiments, the pancreatic fibrosis is associated with pancreatitis {e.g. chronic pancreatitis, acute pancreatitis), cystic fibrosis or pancreatic cancer (e.g. pancreatic ductal adenocarcinoma). Fibrosis of the pancreas and fibrotic diseases of the pancreas are reviewed e.g. in Huang et al., Int J Mol Sci. (2021 ) 22(9): 4970 and Kloppel et al., Virchows Archiv (2004) 445:1 -8, which are hereby incorporated by reference in their entirety.

[0613] The methods of the present invention may be effective to reduce the development or progression of fibrosis. The methods may be effective to prevent progression of fibrosis, e.g. to prevent worsening of, or to slow the rate of development of, fibrosis. In some embodiments, the methods may lead to an improvement in the disease / condition, e.g. a reduction fibrosis or reduction in some correlate of the severity of fibrosis. In some embodiments, the methods may prevent development of a later stage of fibrosis.

[0614] It will be appreciated that diseases / conditions characterised by fibrosis may also exhibit further symptoms / pathologies. For example, a disease / condition characterised by fibrosis may further be characterised by inflammation and / or pathological angiogenesis. In some embodiments, the methods may reduce the development or progression of one or more of: fibrosis, inflammation and pathological angiogenesis. In some embodiments, the methods may be effective to prevent the progression {e.g. to prevent worsening, or the slow the rate of development) of one or more of: fibrosis, inflammation and pathological angiogenesis. In some embodiments, the methods may lead to an improvement in the disease / condition, e.g. a reduction in one or more of: fibrosis, inflammation and pathological angiogenesis; or a reduction in some correlate of the severity of one or more of: fibrosis, inflammation and pathological angiogenesis. In some embodiments, the methods may prevent development of a later stage of one or more of: fibrosis, inflammation and pathological angiogenesis. For example, the methods may reduce / prevent the development / progression of fibrosis and inflammation. The methods may reduce / prevent the development / progression of fibrosis and pathological angiogenesis. The methods may reduce / prevent the development / progression of inflammation and pathological angiogenesis. The methods may reduce / prevent the development / progression of fibrosis, inflammation and pathological angiogenesis.

[0615] Inflammation refers to the bodily response to cell ular / tissue injury, and is characterised by edema, erythema (redness), heat, pain, and loss of function (stiffness and immobility) resulting from local immune, vascular and inflammatory cell responses to infection or injury. The injury may result from e.g. of physical {e.g. mechanical) or chemical insult, trauma, infection, cancer or overactive / aberrant immune responses {e.g. autoimmune disease). Inflammation forms part of the innate immune response, and plays an important physiological role in wound healing and the control of infection, and contributes to the restoration of tissue homeostasis. Inflammation and its role in heath and disease is reviewed e.g. in Chen et al., Oncotarget (2018) 9(6): 7204-7218, which is hereby incorporated by reference in its entirety.

[0616] However, many diseases are associated with an overactive inflammatory response {i.e. excessive inflammation and / or aberrantly activated inflammation), and / or chronic (prolonged) inflammation. Herein, excessive and / or chronic inflammation may be referred to as ‘pathological inflammation’. Pathological inflammation may refer to inflammation which is implicated in ( / '.e. which positively contributes to) the pathology of a disease.

[0617] Inflammation to be treated / prevented in accordance with the present disclosure can be of any tissue / organ of the body. In some embodiments, the inflammation is of the lung {e.g. bronchioles, alveoli), airways {e.g. nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi), heart, kidney, liver, skeletal muscle, blood vessels, eye, skin, pancreas, bowel, small intestine, large intestine, colon, joints, brain, or bone marrow. Inflammation may also occur in multiple tissues / organs at once.

[0618] In some embodiments, inflammation may be of an organ or tissue of the respiratory system, e.g. the lung {e.g. bronchioles, alveoli), or airways {e.g. nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi). In some embodiments, inflammation may be of an organ or tissue of the cardiovascular system, e.g. the heart or blood vessels. In some embodiments, inflammation may be of an organ or tissue of the gastrointestinal system, e.g. of the liver, bowel, small intestine, large intestine, colon, or pancreas. In some embodiments, inflammation may be of the eye. In some embodiments, inflammation may be of the skin. In some embodiments, inflammation may be of an organ or tissue of the nervous system, e.g. the brain. In some embodiments, inflammation may be of the bone marrow. In some embodiments, inflammation may be of the joints. In some embodiments, inflammation may be of an organ or tissue of the urinary system, e.g. the kidneys. In some embodiments, inflammation may be of an organ or tissue of the musculoskeletal system, e.g. muscle tissue. In some embodiments, inflammation may be of the eye. In some embodiments, inflammation may be of an organ or tissue of one or more organ systems.

[0619] As used herein, a disease / condition which is ‘characterised by inflammation’ is a disease / condition in which inflammation is a symptom of the disease / condition. Diseases / conditions characterised by inflammation include, but are not limited to:

[0620] Diseases / conditions affecting the respiratory system, such as sinusitis, rhinitis, pharyngitis, laryngitis, tracheitis, bronchitis, bronchiolitis, pneumonitis, pleuritis and mediastinitis;

[0621] Diseases / conditions affecting the accessory digestive organs such as hepatitis, ascending cholangitis, cholecystitis, pancreatitis (e.g. acute pancreatitis) and peritonitis;

[0622] Diseases / conditions affecting the cardiovascular system such as carditis, endocarditis, myocarditis, cardiogenic shock, pericarditis, vasculitis, arteritis, phlebitis and capillaritis; Diseases / conditions affecting the urinary system such as nephritis, glomerulonephritis, pyelonephritis, ureteritis, cystitis and urethritis;

[0623] Diseases / conditions affecting the nervous system such as encephalitis, myelitis, meningitis, arachnoiditis and neuritis;

[0624] Diseases / conditions affecting the musculoskeletal system such as arthritis, dermatomyositis, soft tissue, myositis, synovitis / tenosynovitis, bursitis, enthesitis, fasciitis, capsulitis, epicondylitis, tendinitis, panniculitis, osteochondritis: osteitis / osteomyelitis, spondylitis, periostitis and chondritis;

[0625] Diseases / conditions affecting the oral cavity and throat such as stomatitis, gingivitis, gingivostomatitis, periodontitis, glossitis, tonsillitis, sialadenitis, parotitis, cheilitis, pulpitis and gnathitis; Diseases / conditions affecting the gastrointestinal system such as esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis, duodenitis, ileitis, caecitis, appendicitis, proctitis and Peutz- Jeghers syndrome;

[0626] Diseases / conditions affecting the skin such as dermatitis, folliculitis, cellulitis and hidradenitis; Diseases / conditions affecting the eye such as dacryoadenitis, scleritis, episcleritis, keratitis, retinitis, chorioretinitis, blepharitis, conjunctivitis and uveitis;

[0627] Diseases / conditions affecting the ear such as otitis externa, otitis media, labyrinthitis and mastoiditis;

[0628] Diseases / conditions of the reproductive system such as oophoritis, salpingitis, endometritis, endometriosis, parametritis, cervicitis, vaginitis, vulvitis, mastitis, orchitis, epididymitis, prostatitis, seminal vesiculitis, balanitis, posthitis, balanoposthitis, chorioamnionitis, funisitis and omphalitis;

[0629] Diseases / conditions of the endocrine system such as insulitis, hypophysitis, thyroiditis, parathyroiditis and adrenalitis;

[0630] Diseases / conditions of the lymphatic system such as lymphangitis and lymphadenitis;

[0631] Cancers, including inflammation-induced and inflammation-associated cancers, such as lung cancer {e.g. lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung cancer), prostate cancer, hematological malignancies {e.g. multiple myeloma), pancreatic cancer (e.g. pancreatic ductal adenocarcinoma), cervical cancer, stomach cancer, oesophageal cancer, head and neck cancer, colorectal cancer, colon cancer, liver cancer {e.g. hepatocellular carcinoma) and bile duct cancer.

[0632] In some embodiments, the inflammation may be acute inflammation. In some embodiments, the inflammation may be chronic inflammation.

[0633] Inflammation can promote angiogenesis {i.e. the growth and development of new blood vessels from existing vasculature), through multiple different pathways as described e.g. in Granger and Senchenkova, ‘Chapter 6: Angiogenesis’, in ‘Inflammation and the Microcirculation’, Morgan & Claypool Life Sciences; 2010. For example, inflammation can lead to hypoxic conditions in inflamed tissue, which in turn upregulates the expression of the potent angiogenic factor vascular endothelial growth factor (VEGF) which induces the growth of new blood vessels. Inflammatory cells such as macrophages, lymphocytes, mast cells and also fibroblasts produce angiogenic factors such as VEGF and FGF. Increased blood flow to inflamed tissue can stimulate angiogenesis through shear stresses on the endothelium of existing vessels, and extravasated plasma proteins, such as fibrinogen products, may also stimulate neovascularisation. In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterised by angiogenesis, {e.g. pathological angiogenesis). In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease / condition characterised by inflammation-induced angiogenesis.

[0634] As used herein, ‘pathological angiogenesis’ refers to angiogenesis {i.e. the growth of new blood vessels from an existing vascular plexus), wherein the angiogenesis contributes to the development and / or progression of a disease. In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease characterised by pathological angiogenesis.

[0635] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is characterised by one or more of fibrosis, inflammation and pathological angiogenesis. In some embodiments, the disease / condition to be treated / prevented is characterised by fibrosis and inflammation. In some embodiments, the disease / condition to be treated / prevented is characterised by fibrosis and pathological angiogenesis. In some embodiments, the disease / condition to be treated / prevented is characterised by inflammation and pathological angiogenesis. In some embodiments, the disease / condition to be treated / prevented is characterised by fibrosis, inflammation, and pathological angiogenesis.

[0636] Fibrosis, inflammation and pathogenic angiogenesis are hallmarks of AMD. This is reviewed in e.g. Helotera and Kaarniranta, Cells (2022) 11 (21 ):3453, which is hereby incorporated by reference in its entirety.

[0637] Administration of the antigen-binding molecules and compositions of the present disclosure is preferably in a ‘therapeutically-effective’ or ‘prophylactical ly-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.

[0638] In some embodiments, the disease / condition to be treated / prevented in accordance with the present disclosure is a disease characterised by tumorigenesis. As used herein, “tumorigenesis” refers to the formation or development of a tumour. Tumorigenesis may involve the transformation of a normal cell(s) into cancerous cell(s). In some embodiments, the disease characterised by tumorigenesis is a cancer.

[0639] In some embodiments, the disease or condition to be treated is a cancer. The cancer may be any cancer as described hereinabove.

[0640] As used herein, a ‘cancer’ may be or comprise any unwanted cell proliferation (or any disease manifesting itself by unwanted cell proliferation), neoplasm or tumor. The cancer may be benign or malignant. The cancer 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, biliary tract, 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, white blood cells.

[0641] Tumors to be treated 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.

[0642] In some embodiments the cancer is a cancer in which LRG1 is pathologically-implicated. That is, in some embodiments the cancer is a cancer which is caused or exacerbated by the expression of LRG1 , a cancer for which expression of LRG1 is a risk factor and / or a cancer for which expression of LRG1 is positively associated with onset, development, progression, severity or metastasis of the cancer. The cancer may be characterised by expression of LRG1 , e.g. the cancer may comprise cells {e.g. cells of tumor tissue) expressing LRG1 . Such cancers may be referred to as being positive for LRG1 . A cancer which is ‘positive’ for LRG1 may be a cancer comprising cells expressing LRG1 . A cancer which is ‘positive’ for LRG1 may overexpress LRG1 .

[0643] 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 LRG1 , for example by quantitative real-time PCR (qRT-PCR). Protein expression can be determined e.g. by antibody-based methods, for example by western blot, immunohistochemistry, immunocytochemistry, flow cytometry, or ELISA.

[0644] In some embodiments, the cancer to be treated is selected from: a cancer comprising cells expressing / overexpressing LRG1 , a solid tumor, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, liver cancer, hepatocellular carcinoma, skin cancer, melanoma, lung cancer, lung adenocarcinoma, squamous cell lung carcinoma, breast cancer, breast carcinoma, ductal carcinoma, colorectal cancer, colorectal carcinoma, colorectal adenocarcinoma, gastric cancer, gastric carcinoma, gastric adenocarcinoma, head and neck cancer, squamous cell carcinoma of the head and neck (SCCHN), ovarian cancer, ovarian carcinoma, ovarian serous adenocarcinoma, kidney cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, renal papillary cell carcinoma, cervical cancer, cervical squamous cell carcinoma, esophageal cancer, esophageal adenocarcinoma, cholangiocarcinoma, uterine cancer, uterine corpus endometrial carcinoma, thyroid cancer, thyroid carcinoma, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma and thymoma.

[0645] In some embodiments, the cancer to be treated is selected from: pancreatic cancer, liver cancer, skin cancer (e.g. melanoma), lung cancer, breast cancer, and colorectal cancer. In some embodiments, the cancer to be treated is selected from: pancreatic cancer, liver cancer, skin cancer (e.g. melanoma) and lung cancer. In some embodiments, the cancer to be treated is selected from: pancreatic cancer, skin cancer (e.g. melanoma), colorectal cancer, and lung cancer. In some embodiments, the cancer to be treated is pancreatic cancer (e.g. pancreatic ductal adenocarcinoma). In some embodiments, the cancer to be treated is skin cancer (e.g. melanoma). In some embodiments, the cancer to be treated colorectal cancer. In some embodiments, the cancer to be treated is lung cancer (e.g. non-small cell lung cancer). The treatment / prevention may be aimed at one or more of: delaying / preventing the onset / progression of symptoms of the cancer, reducing the severity of symptoms of the cancer, reducing the survival / growth / invasion / metastasis of cells of the cancer, reducing the number of cells of the cancer and / or increasing survival of the subject. In some embodiments, treatment with an antigen-binding molecule (e.g. an antigen-binding molecule as described herein) achieves one or more of the following: reducing the survival / growth / proliferation of cells of the cancer, reducing migration / invasion of cells of the cancer, and reducing epithelial to mesenchymal transition (EMT) of cells of the cancer.

[0646] In some embodiments, the disease or condition to be treated is pancreatitis, e.g. acute pancreatitis.

[0647] Acute pancreatitis is a common clinical condition, the incidence of which has been increasing over recent years. Mild acute pancreatitis may be self-limiting and not requiring any treatment, but up to 25% of patients suffer a severe attack and between 30 and 50% of these will die. Most cases are secondary to biliary disease or excess alcohol consumption. Acute pancreatitis is an inflammatory disorder, which develops a complex cascade of immunological events, which not only affect the pathogenesis but also the course of the disease. It will be appreciated that acute pancreatitis is a fibroinflammatory disease according to the present disclosure.

[0648] Acute pancreatitis and the role of inflammation in the disease is reviewed e.g. in Bhatia et al., (2005) 5:132-144, which is hereby incorporated by reference in its entirety.

[0649] The treatment may be aimed at one or more of: reducing the severity of the symptoms of acute pancreatitis, reducing inflammation in acute pancreatitis, and increasing pancreatic regeneration in acute pancreatitis.

[0650] In some embodiments, the disease or condition to be treated is arthritis, e.g. rheumatoid arthritis.

[0651] Rheumatoid arthritis (RA) is a chronic autoimmune and inflammatory condition often affecting symmetrical joints of the body. As the disease progresses, symptoms can spread from smaller joints to weight-bearing joints, such as ankles, knees, and hips. Over time, the joints may deform and eventually lose function. Besides the articular presentation, around 40% of RA patients also experience systemic manifestations affecting all aspects of patients’ organ systems, and these complications are often more fatal to people with RA than those without. The primary goal of RA therapy is to minimize disease activity and control joint damage. However, the disease symptoms persists in a substantial number of patients despite active treatment and a variety of side effects have been reported for existing antirheumatic drugs. Given the limitations of current treatment for RA, there is a need for more effective treatment for RA.

[0652] It will be appreciated that rheumatoid arthritis is a fibroinflammatory disease according to the present disclosure. Rheumatoid arthritis and the role of inflammation in rheumatoid arthritis are reviewed in e.g. Shrivastava & Pandey, Journal of Physiology and Biochemistry (2012) 69:335-347, which is hereby incorporated by reference in its entirety.

[0653] The treatment / prevention may be aimed at delaying / preventing the onset / progression of symptoms of rheumatoid arthritis, reducing the severity of symptoms of rheumatoid arthritis, and / or reducing inflammation in rheumatoid arthritis.

[0654] In some embodiments, the disease or condition to be treated is inflammatory bowel disease (e.g. Crohn’s disease (CD), ulcerative colitis (UC)), In some embodiments, the disease or condition to be treated is Crohn’s disease. In some embodiments, the disease or condition to be treated is ulcerative colitis.

[0655] The treatment / prevention may be aimed at delaying / preventing the onset / progression of symptoms of inflammatory bowel disease, reducing the severity of symptoms of inflammatory bowel disease, and / or reducing inflammation in inflammatory bowel disease.

[0656] In some embodiments, the disease or condition to be treated is associated with diabetes. In some embodiments, the disease or condition to be treated is associated with diabetic vascular complications. Diseases / conditions associated with diabetes include diabetic retinopathy, diabetic macular edema diabetic nephropathy and diabetic wound(s). In some embodiments, the disease or condition to be treated is selected from: diabetic retinopathy, diabetic macular edema, diabetic nephropathy and diabetic wound(s).ln some embodiments, the disease or condition to be treated is selected from: diabetic nephropathy and diabetic wound(s). In some embodiments, the disease or condition to be treated is diabetic nephropathy. In some embodiments, the disease or condition to be treated is a diabetic wound. In some embodiments, the disease or condition to be treated is diabetic retinopathy. In some embodiments, the disease or condition to be treated is a diabetic macular edema.

[0657] Diabetic nephropathy (DN) is a common renal disease associated with long-term diabetes mellitus. Over a lifetime, diabetic nephropathy occurs in approximately 30-35% of patients with type 1 and type 2 diabetes. It is one of the leading causes of end-stage renal disease (ESRD) globally. Chronic high blood sugar levels lead to structural and functional changes in the kidneys. This includes thickening of the glomerular basement membrane, mesangial expansion, and podocyte injury. Its early stage is often asymptomatic, with microalbuminuria as the first detectable sign. With the disease progression, persistent proteinuria (macroalbuminuria), hypertension, and declining glomerular filtration rate (GFR) start to show up. Significant reduction in GFR indicates the advanced stage of DR, which leads to chronic kidney disease (CKD) and eventually ESRD, requiring dialysis or kidney transplantation. With the exception of kidney dialysis and renal replacement therapy, few therapeutic strategies have been found to be effective in treating diabetes nephropathy. Early detection allows time for the intensive treatment of glycaemic control, blood pressure, and other cardiovascular risk factors.

[0658] Diabetic wounds are a serious complication of diabetes, primarily resulting from a combination of poor blood circulation, nerve damage (neuropathy), and elevated blood sugar levels that impair the body's ability to heal. These wounds most commonly manifest as diabetic foot ulcers (DFUs), which affect up to 25% of people with diabetes during their lifetime. Diabetic foot ulcers typically appear as open sores on the feet, especially in pressure-bearing areas like the heel or ball of the foot, and are often painless due to nerve damage, making early detection challenging. Other types of diabetic wounds include venous stasis ulcers, arterial (ischemic) ulcers, pressure injuries, and traumatic wounds — all of which are slow to heal and prone to infection.

[0659] Complications from diabetes also occur in the eye. Diabetic retinopathy (DR) is classically characterized by gradually progressing changes that occur in the microvasculature. These changes include alterations in the retinal permeability, macular edema, retinal ischemia and neovascularization. Diabetic macular edema (DME) is a major cause of vision impairment in diabetic individuals, characterized by fluid accumulation in the macula due to a breakdown of the blood-retinal barrier (BRB).

[0660] The treatment / prevention may be aimed at delaying / preventing the onset / progression of symptoms of a disease / condition associated with diabetes, reducing the severity of symptoms of a disease / condition associated with diabetes, reducing inflammation in a disease / condition associated with diabetes, and / or reducing fibrosis in a disease / condition associated with diabetes. The treatment / prevention may be aimed at delaying / preventing the onset / progression of diabetic nephropathy, reducing the severity of symptoms of diabetic nephropathy, reducing inflammation in diabetic nephropathy, and / or reducing fibrosis in diabetic nephropathy. The treatment / prevention may be aimed at delaying / preventing the onset / progression of a diabetic wound, reducing the severity of symptoms of a diabetic wound, reducing inflammation in a diabetic wound, and / or reducing fibrosis in a diabetic wound.

[0661] In some embodiments, the disease or condition to be treated is an ocular disease or condition described herein (e.g. macular degeneration, Age-related Macular Degeneration (AMD), Geographic Atrophy (‘dry’ or non-exudative AMD), early AMD, early onset macular degeneration (EOMD), intermediate AMD, late / advanced AMD, ‘wet’ (neovascular or exudative) AMD, choroidal neovascularisation (CNV), retinal dystrophy, glaucoma (open-angle or closed-angle), neuromyelitis optica (neuromyelitis optica spectrum disorder (NMOSD)), diabetic macular edema (DME), diabetic retinopathy, proliferative diabetic retinopathy (PDR), polypoidal choroidal vasculopathy, proliferative vitreoretinopathy (PVR), macular edema, drusen formation, Grave's ophthalmopathy, corneal opacification, subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis {e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis and preretinal fibrosis). In such embodiments, the antigen-binding molecule for use in the treatment may be EBC59, EBC60, EBC61 , EBC-1191 , EBC-1192, EBC-1195 or EBC1212. In such embodiments, the antigen-binding molecule for use in the treatment may be EBC59 or EBC1212.

[0662] In some embodiments, the disease or condition to be treated is cancer (e.g. pancreatic cancer). In such embodiments, the antigen-binding molecule for use in the treatment may be EBC61 .

[0663] In some embodiments, the disease or condition to be treated is diabetic nephropathy. In such embodiments, the antigen-binding molecule for use in the treatment may be EBC58, EBC59, EBC60 or EBC61.

[0664] In some embodiments, the disease or condition to be treated is arthritis (e.g. rheumatoid arthritis). In such embodiments, the antigen-binding molecule for use in the treatment may be EBC58.

[0665] In some embodiments, the disease or condition to be treated is inflammatory bowel disease. In such embodiments, the antigen-binding molecule for use in the treatment may be EBC58.

[0666] In some embodiments, the disease or condition to be treated is skin fibrosis. In such embodiments, the antigen-binding molecule for use in the treatment may be EBC58.

[0667] In some embodiments, the disease or condition is characterised by:

[0668] (i) angiogenesis, fibrosis and / or inflammation of the eye;

[0669] (ii) fibrosis and / or inflammation of the pancreas;

[0670] (iii) vascular abnormality, fibrosis and / or inflammation of the joints;

[0671] (iv) vascular abnormality, fibrosis and / or inflammation of the skin;

[0672] (v) fibrosis and / or inflammation of the bowel;

[0673] (vi) vascular abnormality, fibrosis and / or inflammation of the kidney;

[0674] (vii), vascular abnormality, tumorigenesis, fibrosis and / or inflammation of a cancer (e.g. melanoma, pancreatic cancer, colorectal cancer, and lung cancer); or

[0675] (viii) fibrosis and / or inflammation of the lungs.

[0676] As used herein, “vascular abnormality” refers to an irregularity or disorder in the structure or function of blood vessels. For example, vascular abnormality / vascular abnormalities can be found in tumours due to the abnormal growth of blood vessels.

[0677] Administration of the antigen-binding molecules and compositions of the present disclosure may be e.g. parenteral, systemic, topical, intracavitary, intravascular, intravenous, intra-arterial, intramuscular, intrathecal, intraocular, intraconjunctival, intratumoral, subcutaneous, intradermal, oral or transdermal. Administration may be by injection, infusion or ingestion. 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 affected by the 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 {i.e. intravenous / intra- arterial administration) by injection or infusion {e.g. via cannula), or may be administered intraocularly (e.g. by intraocular injection).

[0678] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure may further comprise administering another agent for the treatment / prevention of the relevant disease / condition. Administration of antigen-binding molecules and compositions described herein may be alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated. Simultaneous administration refers to administration with another therapeutic agent together, for example as a pharmaceutical composition containing both agents (combined preparation), or immediately after each other {e.g. within 1 , 4, 6, 8 or 12 hours) and optionally via the same route of administration {e.g. to the same tissue, artery, vein or other blood vessel). Sequential administration refers to administration of one agent followed after a given time interval by separate administration of another 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.

[0679] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure may further comprise administering an antiangiogenic therapy, e.g. a VEGF inhibitor. In some embodiments, therapeutic or prophylactic intervention according to the present disclosure may further comprise administering aflibercept (Eylea).

[0680] Multiple doses of the antigen-binding molecules and compositions may be provided. Multiple doses may be separated by a predetermined time interval, which may be selected to be one of 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1 , 2, 3, 4, 5, or 6 months. By way of example, doses may be given once every 7, 14, 21 or 28 days (plus or minus 3, 2, or 1 days).

[0681] Methods of detection

[0682] The present disclosure also provides the articles of the present disclosure for use in methods for detecting, localizing or imaging LRG1 , or cells expressing LRG1 .

[0683] The antigen-binding molecules described herein may be used in methods that involve detecting binding of the antigen-binding molecule to LRG1 . Such methods may involve detection of the bound complex of the antigen-binding molecule and LRG1 .

[0684] As such, a method is provided, comprising contacting a sample containing, or suspected to contain, LRG1 , and detecting the formation of a complex of the antigen-binding molecule and LRG1 . Also provided is a method comprising contacting a sample containing, or suspected to contain, a cell expressing LRG1 , and detecting the formation of a complex of the antigen-binding molecule and a cell expressing LRG1 .

[0685] Suitable method formats are well known in the art, including immunoassays such as sandwich assays, e.g. ELISA. The methods may involve labelling the antigen-binding molecule, or target(s), or both, with a detectable moiety, e.g. a fluorescent label, phosphorescent label, luminescent label, immuno-detectable label, radiolabel, chemical, nucleic acid or enzymatic label as described herein. Detection techniques are well known to those of skill in the art and can be selected to correspond with the labelling agent.

[0686] Methods comprising detecting LRG1 , or cells expressing LRG1 , include methods for diagnosing / prognosing a disease / condition described herein.

[0687] Methods of this kind may be performed in vitro on a patient sample, or following processing of a patient sample. Once the sample is collected, the patient is not required to be present for the in vitro method to be performed, and therefore the method may be one which is not practised on the human or animal body. In some embodiments, the method is performed in vivo.

[0688] Such methods may involve detecting or quantifying LRG1 and / or cells expressing LRG1 , e.g. in a patient sample. Where the method comprises quantifying the relevant factor, the method may further comprise comparing the determined amount against a standard or reference value as part of the diagnostic or prognostic evaluation. Other diagnostic / prognostic tests may be used in conjunction with those described herein to enhance the accuracy of the diagnosis or prognosis or to confirm a result obtained by using the tests described herein.

[0689] Detection in a sample may be used for the purpose of diagnosis of a disease / condition {e.g. a cancer), predisposition to a disease / condition, or for providing a prognosis (prognosticating) for a disease / condition, e.g. a disease / condition described herein. The diagnosis or prognosis may relate to an existing (previously diagnosed) disease / condition.

[0690] A sample may be taken from any tissue or bodily fluid. The sample may comprise or may be derived from: a quantity of blood; a quantity of serum derived from the individual’s blood which may comprise the fluid portion of the blood obtained after removal of the fibrin clot and blood cells; a tissue sample or biopsy; pleural fluid; cerebrospinal fluid (CSF); or cells isolated from said individual. In some embodiments, the sample may be obtained or derived from a tissue or tissues which are affected by the disease / condition {e.g. tissue or tissues in which symptoms of the disease manifest, or which are involved in the pathogenesis of the disease / condition).

[0691] A subject may be selected for diagnostic / prognostic evaluation based on the presence of symptoms indicative of a disease / condition described herein, or based on the subject being considered to be at risk of developing a disease / condition described herein. The present disclosure also provides methods for selecting / stratifying a subject for treatment with a LRG1 -targeted agent. In some embodiments a subject is selected for treatment / prevention in accordance with the methods of the present disclosure, or is identified as a subject which would benefit from such treatment / prevention, based on detection / quantification of LRG1 , or cells expressing LRG1 , e.g. in a sample obtained from the individual.

[0692] Subjects

[0693] A subject in accordance with the various aspects of the present disclosure may be any animal or human. Therapeutic and prophylactic applications may be in human or animals (veterinary use).

[0694] The subject to be administered with an article of the present disclosure {e.g. in accordance with therapeutic or prophylactic intervention) may be a subject in need of such intervention. The subject is preferably mammalian, more preferably human. The subject may be a non-human mammal, but is more preferably human. The subject may be male or female. The subject may be a patient.

[0695] A subject may have {e.g. may have been diagnosed with) a disease or condition described herein, may be suspected of having such a disease / condition, or may be at risk of developing / contracting such a disease / condition. In embodiments according to the present disclosure, a subject may be selected for treatment according to the methods based on characterisation for one or more markers of such a disease / condition.

[0696] In some embodiments, a subject may be selected for therapeutic or prophylactic intervention as described herein based on the detection of cells / tissue expressing LRG1 , or of cells / tissue overexpressing LRG1 , e.g. in a sample obtained from the subject.

[0697] Kits

[0698] In some aspects of the present disclosure a kit of parts is provided. In some embodiments, the kit may have at least one container having a predetermined quantity of an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell or composition described herein.

[0699] In some embodiments, the kit may comprise materials for producing an antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell or composition described herein.

[0700] The kit may provide the antigen-binding molecule, polypeptide, CAR, nucleic acid (or plurality thereof), expression vector (or plurality thereof), cell or composition together with instructions for administration to a patient in order to treat a specified disease / condition.

[0701] In some embodiments the kit may further comprise at least one container having a predetermined quantity of another therapeutic agent {e.g. as described herein). In such embodiments, the kit may also comprise a second medicament or pharmaceutical composition such that the two medicaments or pharmaceutical compositions may be administered simultaneously or separately such that they provide a combined treatment for the specific disease or condition.

[0702] 5 Kits according to the present disclosure may include instructions for use, e.g. in the form of an instruction booklet or leaflet. The instructions may include a protocol for performing any one or more of the methods described herein.

[0703] Sequence identity 0 As used herein, ‘sequence identity’ refers to the percent of nucleotides / amino acid residues in a subject sequence that are identical to nucleotides / amino acid residues in a reference sequence, after aligning the sequences and, if necessary, introducing gaps, to achieve the maximum percent sequence identity between the sequences. Pairwise and multiple sequence alignment for the purposes of determining percent sequence identity between two or more amino acid or nucleic acid sequences can be achieved in 5 various ways known to a person of skill in the art, for instance, using publicly available computer software such as ClustalOmega (Sbding, J. 2005, Bioinformatics 21 , 951 -960), T-coffee (Notredame et al. 2000, J. Mol. Biol. (2000) 302, 205-217), Kalign (Lassmann and Sonnhammer 2005, BMC Bioinformatics, 6(298)) and MAFFT (Katoh and Standley 2013, Molecular Biology and Evolution, 30(4) 772-780) software. When using such software, the default parameters, e.g. for gap penalty and extension penalty, are preferably 0 used.

[0704] Sequences

[0705] able A

[0706] 93

[0707]

[0708] Table D

[0709] The present disclosure includes the combination of the aspects and preferred features described except where such a combination is clearly impermissible or expressly avoided.

[0710] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0711] Aspects and embodiments of the present disclosure will now be illustrated, by way of example, with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference.

[0712] Throughout this specification, including the claims which follow, unless the context requires otherwise, the word ‘comprise,’ and variations such as ‘comprises’ and ‘comprising,’ will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0713] As used herein, an amino acid sequence or a region of a polypeptide which ‘corresponds’ to a specified reference amino acid sequence or region of a polypeptide has at least 60% (e.g. one of >60%, >65%, >70%, >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 the amino acid sequence / polypeptide / region. An amino acid sequence / region / position of a polypeptide / amino acid sequence which ‘corresponds’ to a specified reference amino acid sequence / region / position of a polypeptide / amino acid sequence can be identified by sequence alignment of the subject sequence to the reference sequence, e.g. using sequence alignment software such as ClustalOmega (Sbding, J. 2005, Bioinformatics 21 , 951 -960).

[0714] It must be noted that, as used in the specification and the appended claims, the singular forms ‘a,’ ‘an,’ and ‘the’ include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from ‘about’ one particular value, and / or to ‘about’ another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by the use of the antecedent ‘about,’ it will be understood that the particular value forms another embodiment.

[0715] Where a nucleic acid sequence is disclosed herein, the reverse complement thereof is also expressly contemplated.

[0716] Methods described herein may preferably be performed in vitro. The term ‘in vitro' is intended to encompass procedures performed with cells in culture whereas the term ‘in vivo’ is intended to encompass procedures with / on intact multi-cellular organisms.

[0717] Brief Description of the Figures

[0718] Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures.

[0719] Figure 1. Graph showing serum LRG1 levels are elevated in nAMD patients with baseline fibrosis and incident fibrosis. Luminex analysis of serum LRG1 levels in nAMD patients without subretinal fibrosis at the baseline and in 12 months (NF), without subretinal fibrosis at the baseline but developing subretinal fibrosis in 12 months (IF), with subretinal fibrosis at the baseline (BF). Data are expressed as mean ± S.E.M of n > 45. Statistical significance was determined by one-way ANOVA. ***P<0.001 , **P<0.01 .

[0720] Figure 2. Graph showing LRG1 levels are induced in the RPE / choroid compartment of the eye of C57BL / 6 mice subjected to laser-induced subretinal fibrosis. qRT-PCR analysis of LRG1 mRNA levels in the RPE / choroid fraction of the eye of C57BL / 6 mice 4, 7, 14, 21 and 35 days following laser treatment. Data are expressed as mean ± S.E.M of n =3.

[0721] Figure 3. Images and graph showing Lrg1 deletion leads to reduced subretinal fibrosis in vivo. Representative images (left) and quantification (right) of total collagen I positive area in wild-type and Lrg1 knockout mice 35 days following the laser treatment. All images are representative. Data are expressed as mean ± S.E.M of n =6. Statistical significance was determined by the student’s t-test. ***P, 0.001. Figure 4. Images showing LRG1 is required for corneal fibrosis. Representative H&E images of cross section of mouse eyes collected from wild-type and Lrg1 mice following the treatment with 1 M NaOH.

[0722] Figure 5. LRC, but not LRR, promotes CTGF expression in HRPC. Representative Western Blot shows an increase in basal and TGFp-induced CTGF expression in LRC treated-HRPCs

[0723] Figures 6A to 6F. Graphs and images showing that LRG1 promotes human RPE cells activation in vitro. (6A) MTS assay demonstrating increased cell survival when RPE is subjected to continuous rhLRGI treatment for 3 days. (6B) Representative IF staining and quantification of Ki67+ RPE following the treatment with rhLRGI . (6C) Representative IF staining of aSMA, Ki67 and DAPI of untreated and LRG1 -treated mouse foetal metatarsal explants. (6D) Representative image and quantification of scratch area as a marker of RPE migratory potential following rhLRGI treatment. (6E). Bar graph demonstrating the fold change in migrated RPE cells across the Transwell following the LRG1 or vehicle treatment. (6F) Representative images and quantification demonstrating the TGFp and TNFa-induced RPE cell contraction following the LRG1 or vehicle treatment. All images are representative. Data are expressed as average ± S.E.M of n=3. Statistical significance was determined by the student’s t-test. *P<0.05, **P<0.01.

[0724] Figures 7A to 7D. LRG1 promotes the expression of fibrotic molecules in human RPE cells cells. (7A) Representative Western Blot and densitometry quantification demonstrating a promoting effect of recombinant human LRG1 (rhLRGI ) on the expression of ECM proteins and the proteins involved in RPE activation. (7B) Quantification of gene transcripts FN, CTGF, TGFB and Coll a1 suggesting increased expression of ECM markers in rhLRGI -treated cells. (7C) Representative IF staining and quantification of fibronectin accumulation in human RPE cultures following rhLRGI treatment. (7D) Representative immunoblots and quantification of pSmad2 / 3 following the treatment of rhLRGI (20 pg / ml) and SB431542 ("10 pM). Data are expressed as average ± S.E.M of n=3. Statistical significance was determined by the student’s t-test. *P<0.05, **P<0.01 , ***P<0.001 .

[0725] Figure 8. Images showing Mouse species cross-reactivity of LRG1 monoclonal antibodies using mouse liver and retina extracts by immunoblots. Recombinant human LRG1 (rhLRGI ) is used as positive control.

[0726] Figure 9. Graphs showing binding kinetics of chimerised EBC-58, EBC-59, EBC-60 and MGZ against native LRG1 protein measured by BLI. Recombinant LRG1 protein was immobilised on ARG2 biosensors. Ligand-loaded sensors were incubated with different concentrations of analyte in BLI buffer (100, 50, 25, 12.5 nM of LRG1 antibodies). The concentration of each curve is orientated as top-down, with top concentration 100 nM presented at the top, followed by 2-fold serial dilutions concentrations down. Binding curves were fitted using a 1 :1 binding model, and fitted lines are shown in red. Figure 10. Peptide mapping of chimeric EBC-58, EBC-59, EBC-60, EBC-61 and MGZ antibodies by ELISA binding assay using biotinylated 15-mer overlapping peptides covering LRR6 region (peptide 1 - 3) and LRR8 and LRRCT regions (peptide 4 -19).

[0727] Figure 11. Graphs showing chimeric LRG1 blocking antibodies inhibit TGFp-induced fibrotic gene expression in RPE cells. RT-PCR analysis of fibrotic gene expression in RPE cells subjected to the treatment rhLRGI with or without the presence of LRG1 blocking antibodies EBC-59, EBC-60, and EBC- 61 (n=3 except for CTGF). Statistical significance was determined by One-Way ANOVA with *p < 0.05, **P< 0.01 , and *** P< 0.001 .

[0728] Figure 12. Images and graph showing that chimeric LRG1-blocking antibodies inhibit TGFp-induced RPE cell migration. Representative images (left) and quantification (right) of RPE cell migration over Transwell following the treatment with TGFp with or without the presence of LRG1 blocking antibodies EBC59, EBC60 and EBC61 . Data are expressed as average ± S.E.M of n=6. Statistical significance was determined by one way ANOVA. *P<0.05, **P<0.01 , ***P<0.001 .

[0729] Figure 13. Images and graph showing chimeric LRG1-blocking antibodies inhibit TGFp-induced fibronectin expression in RPE cells. Representative immunofluorescence images (left) and quantification (right) of fibronectin expression in RPE cells following the treatment with TGFp with or without the presence of LRG1 blocking antibodies EBC-59, EBC-60, and EBC-61 . Data are expressed as average ± S.E.M of n=3. Statistical significance was determined by one-way ANOVA. ***P<0.001 .

[0730] Figure 14. Images and graph showing chimeric LRG1 -blocking antibodies inhibit LRG1 -induced RPE cell proliferation. Representative immunofluorescence images (left) and quantification (right) of Ki67 positive RPE cells following the treatment with rhLRGI with or without the presence of LRG1 blocking antibodies EBC-59, EBC-60, and EB-C61 . Data are expressed as average ± S.E.M of n=3. Statistical significance was determined by one way ANOVA. *P<0.05, **P<0.01 ,***P<0.001 .

[0731] Figure 15. Images and graph showing chimeric LRG1 -blocking antibodies inhibit metatarsal angiogenesis and fibrosis. Representative immunofluorescence images (left) and quantification (right) of CD31 positive vessel outgrowth from metatarsal bones following the treatment with rhLRGI with or without the presence of LRG1 blocking antibodies EBC-59, EBC-60, and EBC-61 . Data are expressed as average ± S.E.M of n=6 (for CD31 staining) and n=3 (for CD31 and aSMA co-staining). Statistical significance was determined by one way ANOVA. *P<0.05, **P<0.01 ,***P<0.001 .

[0732] Figures 16A and 16B. Images and graphs showing chimeric LRG1 -blocking antibodies inhibit laser- induced CNV and subretinal fibrosis. (16A) Representative images (left) and quantification (right) of fundus fluorescein angiography following the treatment with LRG1 blocking antibodies EBC-59, EBC-60, and EBC- 61 . (16B) Representative immunofluorescence images (left) of aSMA and Collagen I stained RPE flatmount following the treatment with LRG1 blocking antibodies EBC-59, EBC-60, and EBC-61 . Quantification of total Collagen 1 positive subretinal fibrosis area (right). Data are expressed as average ± S.E.M of n=6. Statistical significance was determined by one-way ANOVA. *P<0.05, **P<0.01 , ***P<0.001 . Figure 17. Graphs showing binding kinetics of EBC-59 (parental chimeric mAb) and its humanised variants (EBC-1191 , EBC-1192, EBC-1196, EBC-1199, EBC-1200, EBC-1212) to human LRG1 protein as determined by BLI. Recombinant LRG1 protein was immobilised on ARG2 biosensors. Ligand-loaded sensors were incubated with different concentrations of analyte in BLI buffer (100, 50, 25, 12.5 nM of LRG1 antibodies). The concentration of each set of curves is orientated as top-down, with top concentration 100 nM presented at the top, followed by 2-fold serial dilutions concentrations down. Binding curves were fitted using a 1 :1 binding model, and fitted lines are shown in red.

[0733] Figure 18A and 18B: Images and graphs showing the inhibitory effect of humanised variants of EBC59 (EBC1191 , EBC1192, EBC1195, EBC1196, EBC1199, EBC1200, and EBC1212) on RPE cell proliferation. Representative immunofluorescence images (18A) and quantification (18B) of Ki67+ (proliferating RPE cells) to nuclei (total RPE cell count). Data are expressed as average ± S.E.M of n=3. Statistical significance was determined by one-way ANOVA. L: recombinant LRG1 ; n.s: not significant; *P<0.05, ****P<0.0001

[0734] Figure 19. Humanized LRG1 antibody inhibits laser-induced choroidal neovascularization (CNV) in mice in a dose-dependent manner. Representative FFA and IF images (top) and quantification (bottom) demonstrating a dose-dependent inhibition of laser-induced CNV in mice. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA. *P<0.05, **P<0.01 , ****P<0.0001

[0735] Figure 20. Representative FFA images demonstrating an immediate and prolonged anti-angiogenic effect of humanised LRG1 antibody EBC1212 either on its own or in combination with the standard of care Eylea, in comparison to Eylea mono-therapy in Kimba mice.

[0736] Figures 21 A to 21 B. LRG1 blocking antibodies demonstrated a comparative anti-angiogenic efficacy and an additional anti-fibrotic property to the standard of care VEGF inhibitor, Avastin. (21 A) HREC Matrigel tube formation assay demonstrated potent inhibitory efficacy of parental (EBC59) and humanized LRG1 blocking antibody (EBC1212), which is comparable to the standard of care Avastin. (21 B) Scratch assay showed that EBC59 and EBC1212 but not Avastin inhibit the migration of RPE cells. All images are representative. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA. *P<0.05, **P<0.01 , ***P<0.001 , ****P<0.0001 .

[0737] Figure 22. Representative IF images (top) and quantification (bottom) of Collagen I staining and psOCT demonstrating a dose-dependent inhibition of laser-induced subretinal fibrosis in mice by humanized LRG1 antibody EBC1212. All images are representative. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA. *P<0.05, **P<0.01 .

[0738] Figure 23. Representative IF images (top) and quantification (bottom) of Fn, aSMA and IB4 I staining demonstrating a comparable anti-angiogenic efficacy of EBC59 and EBC1212 to a comparator LRG1 blocking antibody EBC78 and the standard of care. However, EBC59 and EBC1212 offer an additional antifibrotic efficacy. EBC146 serves as a negative IgG control. All images are representative. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA. *P<0.05, **P<0.01 , ***P<0.001 , ****P<0.0001 .

[0739] Figure 24. Representative IF images (top) and quantification (bottom) of Fn, aSMA and IB4 I staining demonstrating an additive anti-angiogenic efficacy and an additional anti-fibrotic benefit of EBC1212 to the standard of care Eylea as a combination treatment with Eylea. All images are representative. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA, **P<0.01 , ***P<0.001 , ****P<0.0001 .

[0740] Figure 25. Representative IF images (top) and quantification (bottom) of Fn, aSMA and IB4 I staining demonstrating the ability of EBC1212 to slow down the progression of the laser-induced subretinal fibrosis. All images are representative. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA, *P<0.05 **P<0.01 , ***P<0.001 , ****P<0.0001.

[0741] Figure 26. Immunofluorescence staining images of mouse pancreatic tissue showing colocalization of LRG1 and CD31. For the staining, paraffin (6pm) or cryosections (8pm) were subjected to antigen retrieval using sodium citrate buffer before being stained with primary antibodies against LRG1 , CD31 and AN2 followed by staining with DAPI, Alexa 488, Alexa 594 and Alexa 657 secondary antibodies.

[0742] Figures 27A to 27C. Graphs and images showing an increase LRG1 expression in human patients with acute pancreatitis. (27A) Concentration of LRG1 in serum of patients with acute pancreatitis and a healthy control. (27B) Correlation between serum LRG1 concentration and inflammation marker C- reactive protein. (27C) Immunofluorescence staining for LRG1 in CD31 + vascular cells, Amy+ (amylase) acinar cells and MPO+ (myeloperoxidase) inflammatory cells in human acute pancreatis (Figure 27C).

[0743] Figure 28. Schematic showing the experimental protocol using cerulean to induce acute pancreatitis in mice.

[0744] Figures 29A and 29B. Graphs showing an increase in serum LRG1 concentration (29A) and pancreatic LRG1 mRNA expression (29B) in the caerulein-induced acute pancreatitis mouse model. mRNA quantity was determined by quantitative RT-PCR. The expression level of LRG1 was normalized to RPLPO, and relative gene expressions were calculated using standard 2AACT.

[0745] Figure 30A and 30B. Graphs and images showing an increase in expression of LRG1 protein in pancreatic tissue during cerulean-induced acute pancreatitis. (30A) Western Blot analysis of pancreatic LRG1 protein levels following the onset of cerulean-induced acute pancreatitis. (30B) Immunofluorescence staining for LRG1 , CD31 , amylase (AMY) and myeloperoxidase (MPO). Staining was performed 24 hours after caerulein treatment. Figure 31 A to 31 C. Graphs showing LRG1 expression during acute pancreatitis. (31 A) Pancreatic LRG1 expression as measured by qRT-PCR following bone marrow transplantation and caerulein induction of acute pancreatitis. Three donor / recipient combinations were evaluated: wild-type to wild-type, LrgT / _knock-out to wild-type, and wild-type to LrgTAknock-out. Acute pancreatitis was induced in recipient mice 5 weeks post-transplantation. (31 B) qRT-PCT analysis of LRG1 mRNA, and Western Blot analysis of LRG1 protein in isolated acinar cells following caerulein induction of acute pancreatitis.

[0746] Figures 32A and 32B. Images and graphs showing accelerated pancreatic recovery in the LrgT / _knockout compared to Wild-type. (32A) Immunofluorescence staining of proliferation marker Ki67 and AMY in acinar cells from wild-type and Lrg1 / - knock outs following caerulein induction of acute pancreatitis. (32B) Expression of cyclins CCNB, CCND1 and CCNE 'n the pancreas as determined by qRT-PCR in wild-type and LrgK knock out mice following caerulein induction of acute pancreatitis.

[0747] Figures 33A to 33G. Images and graphs showing that acinar cell function is regulated by LRG1 through AKT-mediated CCKAR expression. (33A and 33B) qRT-PCT analysis of pancreatic CCKAR mRNA expression, and Western Blot analysis of pancreatic CCKAR protein expression in wild-type and Lrgl1- knock out mice. (33C) Western blot analysis of TGFp associated non-canonical signalling proteins AKT, ALK5 and ERK in wild-type and Lrgl1- knock out mice. (33D) Western blot analysis of AKT and ALK5 in acinar cells in wild-type, Lrgl1- knock out and Lrgl1- knock out treated with recombinant LRG1 (Lrgl1-*- rhLRGI ). (33E) qRT-PCT analysis of CCKAR mRNA expression, and Western Blot analysis of CCKAR protein expression in isolated acinar cells of wild-type, Lrgl '- and Lrgl '- treated with rhLRGI . (33F) Western Blot analysis of AKT and ALK5 in Lrgl1- acinar cells following treatment with ALK5 / Smad2 / 3 inhibitor SB431542 and AKT inhibitor MK2206. (33G) qRT-PCT analysis of CCKAR mRNA expression, and Western Blot analysis of CCKAR protein in Lrgl1- acinar cells following treatment with inhibitors SB431542 and MK2206.

[0748] Figures 34A to 34H. Images and graphs showing that an LRG1 blocking antibody can alleviate the effects of acute pancreatitis. (34A) Schematic outlining the experimental protocol to test the effect of LRG1 blocking antibodies on acute pancreatitis induced by caerulein. (34B) Western blot analysis of TGFp associated signalling proteins AKT and ALK5 following caerulein induced acute pancreatitis and subsequent treatment with LRG1 blocking antibody or control IgG. (34C) qRT-PCR analysis of CCK1R mRNA, and Western Blot analysis of CCK1 R protein, following caerulein induced acute pancreatitis and subsequent treatment with LRG1 blocking antibody or control IgG. (34D) H&E staining and pathology evaluation of pancreatic tissue following caerulein induced acute pancreatitis and subsequent treatment with LRG1 blocking antibody or control IgG. (34E, 34F) qRT-PCR analysis of anti-inflammatory NFKBIA and 1110 mRNA (34E) or amylase (AMY2) and cell cycle markers (CCNB and CCNE) (34F) following caerulein induced acute pancreatitis and subsequent treatment with EBC-103 or control IgG. (34G) Immunofluorescence staining for proliferation marker Ki67 and amylase (AMY) following caerulein induced acute pancreatitis and subsequent treatment with LRG1 blocking antibody or control IgG. All images are representative. Data are expressed as average ± S.E.M of n>6. Statistical significance was determined by one-way ANOVA. *P<0.05, **P<0.01 , ****P<0.0001 Figure 35A to 35C. Graphs and images showing upregulation of LRG1 in PDAC cells. (35A; left panel) RNA sequencing expression data from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) project showing differential mRNA expression of LRG1 in PDAC compared to normal pancreas tissue. (35A; right panel) Data extracted from the Clinical Proteomic Tumour Analysis Consortium (CPTAC) database to show LRG1 protein abundance in normal versus human PDAC tumour tissues. (35B) Kaplan-Meier survival analyses of PDAC patients with differential expression of LRG1 obtained from the Human Protein Atlas database. (35C) Immunofluorescence staining of PDAC low- and high-grade tumour ductal cells for LRG1 and KRT19 to establish colocalization.

[0749] Figure 36A to 36H. Images and graphs showing LRG1 promotes tumour growth, epithelial-mesenchymal transition (EMT) and metastasis in-vivo. (36A) Schematic detailing experimental protocol to introduce PDAC cells transformed with a pCDH-LRG1 expression vector into a mouse model. (36B) Analysis of weight and volume of PDAC tumours expressing either a control or pCDH-LRG1 expression vector. (36C) Representative immunofluorescence images (left) and quantification (right) of Ki67 positive cells in PDAC tumours overexpressing LRG1 (pCDH-LRG1 ) and a vector control (pCDH). (36D) qRT-PCR analysis of cyclin Ccnd mRNA expression in PDAC tumours overexpressing LRG1 (pCDH-LRG1 ) and a vector control (pCDH). (36E) Semi-quantitative scoring of neoplastic cell and neoplastic nuclear pleomorphisms following hematoxylin-eosin staining and histopathological grading of in PDAC tumours overexpressing LRG1 (pCDH-LRG1 ) and a vector control (pCDH). In general, a score of 0 indicated that there were no abnormalities detected; 1 : minimal; 2: mild; 3: moderate; 4: marked; 5: severe. For pleomorphism, a semi- quantitative score of 1 -fold, 2-fold, or 3-fold change compared to normal cells was determined. (36F) Staining of PDAC tumour tissue overexpressing LRG1 (pCDH-LRG1 ) (right) or a vector control (pCDH) (left) for mesenchymal N-cadherin. (36G) qRT-PCR analysis of cadherin (CDH2) mRNA expression in PDAC tumours overexpressing LRG1 (pCDH-LRG1 ) or a vector control (pCDH). (36H) Analysis of liver weight and metastatic area of livers from mice having a PDAC tumour overexpressing LRG1 (pCDH- LRG1 ) or a vector control (pCDH).

[0750] Figure 37A to 37F. Images and graphs showing LRG1 promotes cell proliferation, migration and invasion in-vitro. (37A) Cell viability of human PDAC cells (PANC-1 ) transformed with either an empty control vector (pcDNA) or an LRG1 expressing vector (pLRG1 ) over 3 days as a measure of cell proliferation. (37B) Cell viability of PDAC cells (PANC-1 ) treated with siRNA (siLRGI ) to knock down LRG1 expression, or a control siRNA (siCtrl) over 3 days as a measure of cell proliferation. (37C and 37D) Representative images of cell migration and invasion assays of PDAC cells transformed with pLRG1 or pcDNA control (37C), and siLRGI or siCtrl control (37D). (37E) Western Blot analysis and quantification of proliferation marker cyclin D (CCND) protein and EMT marker N-Cadherin (CDH2) protein expression in PDAC cells (PANC-1 ) transformed with an empty control vector (pcDNA) or an LRG1 expressing vector (pLRG1 ). (37F) Western Blot analysis and quantification of proliferation marker cyclin D (CCND) protein and EMT marker N-Cadherin (CDH2) protein in PDAC cells (PANC-1 ) treated with siRNA (siLRGI ) to knock down LRG1 expression, or a control siRNA (siCtrl).

[0751] Figure 38A to 38E. Images and graphs showing LRG1 promotes tumorigenesis via EGFR / ErbB / AKT signalling. (38A) Dot blot profiler assay measuring the protein expression of EGFR ligand amphiregulin in mouse KPC cells overexpressing LRG1 (pCDH-LRG1 ) and control KPC cells (pCDH). (38B) qRT-PCR analysis of amphiregulin (Areg) mRNA expression in KPC cells (left) and tumour (right) overexpressing LRG1 (pCDH-LRG1 ) and control KPC cells (pCDH). (38C) Dot blot profiler assay measuring expression of EGFR, ErBb2, AKT and GSK signalling transducers in KPC cells overexpressing LRG1 (pCDH-LRG1 ) and control KPC cells (pCDH). (38D) Western Blot analysis of EGFR, ErBb2, PDK1 , AKT and GSK signalling protein expression in KPC cells overexpressing LRG1 (pCDH-LRG1 ) and control KPC cells (pCDH). (38E) Western Blot analysis of EGFR / ErbB / AKT associated signalling proteins in KPC cells overexpressing LRG1 following treatment with inhibitors Tucatinib (0.5pM) and MK2206 (10pM) or DMSO control. (38F) Cell viability assay results of mouse KPC cells overexpressing LRG1 and treated with Tucatinib or MK2206 or DMSO control as a measure of cell proliferation. (38G) Representative images and quantification of cell migration and invasion assays of KPC cells overexpressing LRG1 treated with Tucatinib, MK2206, or DMSO control.

[0752] Figure 39A to 39C. Graphs and images showing administration of LRG1 neutralizing antibody (EBC-61 ) inhibits tumour growth in vitro and in vivo. (39A) Cell viability assays of KPC cells overexpressing LRG1 (KPC pCDH-Lrg1 (also referred to herein as “KPC pLrgl )) following treatment with anti-LRG1 antibody EBC-61 or an isotype control antibody as a measure of cell proliferation. (39B) Representative images of cell migration assays measuring migration of KPC cells overexpressing LRG1 following treatment with EBC-61 or an isotype control antibody. Immunofluorescence staining (left) and quantitative analysis (right) of migrated or invaded of control IgG or LRG1 antibody-treated pCDH-LRG1 KPC cells, Scale bar: 100pm. (39C) Western blot analysis and quantification of CCND and CDH2 protein expression in KPC cells overexpressing LRG1 following treatment with EBC-61 or an lgG1 isotype control antibody. (39D) Representative images of KPC pLrgl pancreatic tumours in IgG- or EBC 61 - treated wild-type mice. (39E) Endpoint KPC pLrgl tumour volumes following treatment with IgG- or EBC 61 -antibodies. All images are representative. Data are presented as the mean ± s.e.m. Significance was determined by unpaired, two-tailed Student’s t-test of n > 3 mice or independent experiments; *: p< 0.05, **: p< 0.01 .

[0753] Figure 40A and 40B. LRG1 is highly expressed in rheumatoid arthritis (RA) patients. (40A; left panel) ELISA analysis of LRG1 expression in serum of rheumatoid arthritis (RA) and age-matched osteoarthritis (OA) patients (n=3). (40A; middle panel) Quantitative real-time PCR analysis of LRG1 in synovial tissues collected from RA and age-matched OA patients (n=3). (40A; right panel) ELISA analysis of LRG1 expression in synovial fluid of RA and age-matched OA patients (n=3). (40B) Western blot and quantitative real-time PCR (qRT-PCR) analysis of LRG1 expression in primary fibroblast-like synoviocytes (FLS) isolated from RA and age-matched OA patients (n=3). Data are expressed as mean ± SEM. Statistical significance was determined by two-tailed, unpaired student’s t-test. *P<0.05.

[0754] Figure 41 A to 411. LRG1 regulates the function of multiple cell types in the synovium. (41 A) Representative images of immunofluorescence staining of adherent junction protein VE-cadherin in EC subjected to the treatment with rhLRGI . Paracellular gaps and disrupted intercellular contacts are marked by arrows. DAPI is used to label the nucleus and F-Actin highlights the stress fibres. (41 B) Mile’s assay shows that Lrg1 - / - mice are resistant to VEGF-induced vessel permeability. (41 C) Representative images (left) and quantification (right) of Ki67 positive synovial endothelial cell (EC) in control and rhLRGI treatment conditions. (41 D) Representative images (left) and quantification (right) of the number of synovial ECs migrated across the Transwell in control and rhLRGI treatment conditions. (41E) Representative images (left) and quantification (right) of synovial EC tube formation in control and rhLRGI treatment conditions. (41F) Representative images (left) and quantification (right) of the number of neutrophils adhered to the synovial ECs in control and rhLRGI treatment conditions. (41G) Representative images (left) and quantification (right) of the number of neutrophils migrate across the synovial ECs in control and rhLRGI treatment conditions. (41 H) FLS proliferation as demonstrated by MTS assay of control and rhLRGI treatment conditions. (411) qRT-PCR analysis of the MMP9 gene expression in FLS in control and following the treatment of rhLRGI . All images are representative. Data are expressed as average ± SEM, n>3. Statistical significance was determined by two-way ANOVA or two-tailed, unpaired student’s t-test, *p<0.05, **p<0.01 , ***p<0.001

[0755] Figure 42A and 42B. Collagen-induced arthritis is alleviated in Lrg1- / - mice. Wild-type and Lrg1- / - mice were subjected to CIA. (42A) Representative images on day 53 post collagen treatment (left) and clinical scoring (right) showing macroscopic joint symptoms in wild-type or Lrg1- / - mice post collagen treatment. Arrow indicates swelling of affected limbs. Severity scores were based on the symptoms in WT (n=4) and Lrg1- / - mice (n=5). (42B) Representative H&E images of hind paw joints of wild-type and Lrg1- / - mice at day 60 post collagen treatment. Data are expressed as average ± SEM. Statistical significance was determined by two-tailed, unpaired student’s t-test, *p<0.05.

[0756] Figure 43A to 43E. LRG1 inhibition in Melanoma. (43A) Representative images of B16F10 tumours dissected from IgG- or anti-LRG1 antibody treated wild-type mice. (43B) B16F10 tumour volume over time in IgG- or anti-LRG1 antibody-treated wild-type mice. (43C) Endpoint tumour volume following treatment with IgG- or anti-LRG1 antibody. (43D) qRT-PCR analysis of Ki67 mRNA levels in tumours treated with IgG- or anti-LRG1 antibody. (43E) qRT-PCR analysis of N-Cadherin mRNA levels of in tumours treated with IgG- or anti-LRG1 antibody. Data are presented as mean ± s.e.m. Significance was determined by one-way ANOVA followed by Dunnett’s multiple comparison test n > 3 mice; *: p < 0.05, **: p < 0.01.

[0757] Figure 44A to 44F. LRG1 inhibition in CRC. (44A) Representative images of HT-29 tumours dissected from IgG- or EBC-treated Balb / c nude mice. (44B) HT-29 tumour volume over time in IgG- or antibody- treated Balb / c nude mice. (44C) Endpoint tumour volume following treatment with IgG- or antibodies. (44D) MTS assay demonstrating the cell viability of HT-29 CRC cells treated with control IgG or LRG1 antibodies, EBC 103, EBC 107 and EBC78 over 3 days. (44E) Day 3 endpoint MTS assay results demonstrating cell viability of HT-29 CRC cells treated with control IgG or LRG1 antibodies, EBC 103, EBC 107 and EBC78. (44F) Representative Western blot images showing increased E-cadherin (reduced epithelial -mesenchymal transition) and reduced Cyclin D1 (reduced cell proliferation) in EBC103 treated HT-29 cells compared to IgG- treated controls. Data are presented as mean ± s.e.m. Significance was determined by one-way ANOVA followed by Dunnett’s multiple comparison test n > 3 mice or independent repeats. Figure 45A to 45C. LRG1 inhibition in primary human glomerular mesangial cells. (45A) Representative Western blot and densitometry analysis of (45B) CTGF and (45C) FN1 protein levels following treatment with IgG- or EBC antibodies in the presence of high glucose and TGF-01 . All images are representative. Data are presented as the mean ± s.e.m. Significance was determined by one-way ANOVA followed by Dunnett’s multiple comparison test n > 3 independent repeats.

[0758] Figure 46. Metatarsal angiogenesis assay with anti-LRG1 antibodies. Representative immunofluorescence images of CD31 -stained fetal metatarsal bone explants of vascular density normalized to the explant size. Data are expressed as mean ± S.E.M., n=6 / group. Statistical significance was determined by one-way ANOVA, * p<0.05, ** p<0.01 , *** p<0.001 .

[0759] Figure 47. Metatarsal angiogenesis assay with anti-LRG1 antibodies. Quantitative analysis of vascular density normalized to the explant size. Data are expressed as mean ± S.E.M., n=6 / group. Statistical significance was determined by one-way ANOVA, * p<0.05, ** p<0.01 , *** p<0.001 .

[0760] Figure 48A to 48E. (48A) Schematic of DSS-induced acute colitis model and LRG1 antibody treatment scheme. (48B) Tabulated daily body weight loss and (48C) disease activity index (DAI). (48D) Representative endpoint gross necropsy images of whole intestinal tract and (48E) tabulated colon length at endpoint, day 10. Data are expressed as mean ± S.E.M., n=6 / group. Statistical significance was determined by one-way ANOVA, * p<0.05, ** p<0.01 , *** p<0.001 .

[0761] Figures 49A to 49B. (49A and B) Histological scoring of disease severity following LRG1 -neutralizing antibody treatment. (49A) Representative H&E staining of treatment groups. (49B) Tabulated histological scores evaluating epithelial morphology, immune cell infiltration and global architecture. Data are expressed as mean ± S.E.M., n=6 / group. Statistical significance was determined by one-way ANOVA, * p<0.05, ** p<0.01 , *** p<0.001.

[0762] Figure 50A to 50E. In vivo validation of LRG1 -blocking antibodies in CAIA mice model. (A) Timeline of CAIA induction, LPS booster and LRG1 -neutraliing antibody treatment scheme. (B) Daily body weight tracking and (C) combined clinical scores of all four limbs of experimental mice. (D) Clinical score for CAIA mice treated with LRG1 blocking antibodies and isotype control at day7 post-induction (Man- Whitney test). (E) Percentage reduction in grip strength of CAIA mice treated with LRG1 blocking antibodies and isotype control (one-way ANOVA). Data are expressed as mean ± S.E.M, n = 8 / group, statistical significance presented as *p < 0.05, “ p < 0.01 , p < 0.001 .

[0763] Figures 51 A to 51 D Representative histological staining. Hematoxylin and Eosin (H&E) staining of sagittal sections (51 A) and transverse sections (51 B) of the hind paws of CAIA mice treated with LRG1 - blocking antibodies or isotype control (n=8 / group). White stars indicate areas of synovial inflammation; black triangles indicate areas with bone inflammation and dotted lines indicate the regions of bone erosion. Representative Safranin O / Fast green staining of sagittal sections (51 C) and transverse sections (51 D) of the hind paws of CAIA mice treated with LRG1 -blocking antibodies or isotype control (n=8 / group). White block arrows indicate areas of destaining of superficial cartilage while black pointed arrows indicate areas with erosion of superficial cartilage.

[0764] Figures 52A to 52C. LRG1 inhibition in NSCLC. (52A) Representative images of LL / 2 tumours dissected from IgG- or EBC-treated wild-type mice. (52B) LL / 2 tumour volume over time in IgG- or EBC-treated wild-type mice. (52C) Endpoint tumour volume following treatment with IgG- or EBC antibodies. Data are presented as mean ± s.e.m. Significance was determined by one-way ANOVA followed by Dunnett’s multiple comparison test n > 3 mice; p < 0.01 , ***: p < 0.001 .

[0765] Figures 53A to 53M. LRG1 inhibition in AR mouse model of DN. (53A) qRT-PCR analysis of Lrg1 mRNA levels in the kidneys of wild-type and AR mice. (53B) Changes in urinary albumin excretion rates (UAE) in untreated or EBC-107 treated AR mice from 12-weeks to 18-weeks. (53C) Urinary albumin excretion rates (UAE) in untreated or EBC-107 treated AR mice at 18-weeks endpoint. (53D) Representative Masson’s Trichrome staining of glomerular and tubulo-interstitial regions in untreated and EBC-107 treated AR mouse kidney sections. (53E) Quantitative analysis of trichrome positive area over total kidney area in untreated and EBC-107 treated AR mice. (53F) Representative Western blot and densitometry analysis of (53G) COL1 and (53H) COL4 protein levels following treatment with EBC-107 compared to untreated counterparts. qRT-PCR analysis of (53I) Mmp2, (53J) Kimi, (53K) Ccl2, (53L) Ccl5, (53M) 116. Data are presented as mean ± s.e.m. Significance was determined by unpaired two-tailed Student’s t-test of n > 3 mice; *: p < 0.05, **: p < 0.01 .

[0766] Figure 54A and 54B. Representative experimental timeline of the mouse model of bleomycin-induced skin fibrosis and schematic illustrations of dorsal subdermal injection sites (A). Weekly measurements of skin-fold thickness (B). Data are expressed as mean ± S.E.M, n = 3 to 5 / group.

[0767] Figures 55A and 55B. Representative Mason’s Trichrome staining images of full-thickness skin sections, at 10X magnification and 40X magnification, with scale bar indicative of 200um and 400umm respectively (A). Representative immunoblots and densitometric analysis of classic ECM markers COL1 A1 , FN1 and aSMA in LRG1 -blocking antibodies treated skin. Data expressed as mean ± S.E.M, n = 3 to 5 / group.

[0768] Figures 56A and 56B. Schematic illustration of experimental timeline involving STZ-induced diabetes, 8mm full-thickness excisional wound, and subsequent delivery of LRG1 -blocking antibodies to promote chronic wound healing (A). Representative illustration indicating site of dorsal cutaneous wound creation and initial wound size before treatment.

[0769] Figures 57A and 57B. Representative wound images demonstrating time-dependent wound healing trajectory and physical changes of wound appearance (B). Quantitative analysis of wound closure over experimental period (C). Data are expressed as mean ± S.E.M, n = 3 to 5 / group. Examples

[0770] Example 1 : LRG1 level is closely associated with subretinal fibrosis in both disease animal model and humans

[0771] LRG1 was previously shown to be expressed at high levels in the vitreous of human patients with neovascular age-related macular degeneration (nAMD). To establish the association between LRG1 and subretinal fibrosis, serum LRG1 levels were analysed in human nAMD patients without subretinal fibrosis at the baseline and in 12 months (NF), without subretinal fibrosis at the baseline but develop subretinal fibrosis in 12 months (IF), and with subretinal fibrosis at the baseline (BF). Results showed that serum LRG1 levels are significantly higher in IF and BF patients compared to that in patients without fibrosis at all (Figure 1 ). This observation was further confirmed in an animal model of laser-induced choroidal neovascularization (CNV). In this model, subretinal fibrosis becomes apparent on day 35 post-laser injury. Results showed that the expression of LRG1 in the retinal pigmented epithelium (RPE) / choroidal fraction of the eye is significantly induced at day 35 post-laser (Figure 2). Together, these data demonstrate a close association between LRG1 and subretinal fibrosis in both diseased mice and humans.

[0772] Example 2: Lrg1 deletion leads to reduced subretinal and corneal fibrosis

[0773] Following the establishment of the association between elevated LRG1 levels and subretinal fibrosis in human patients and disease mice, the impact of Lrg1 deletion in subretinal fibrosis was investigated. Wild-type and Lrg1 knockout mice were subjected to laser treatment. RPE flatmount collected from wildtype and Lrg1 knockout mice 35 days after the laser treatment were subjected to immunofluorescence staining with a fibrotic marker collagen I. Results showed that the total area of collagen l-positive lesions is significantly reduced in Lrg1 knockout mice as compared to that in wild-type controls (Figure 3), demonstrating that LRG1 is required for subretinal fibrosis in vivo.

[0774] To further expand the cause-effect relationship between LRG1 and ocular fibrosis at the anterior of the eye such as corneal, the extent of the corneal fibrosis was examined in wild-type and Lrg1 mice following the exposure to 1 M NaOH. Results showed that there is a significant reduction in corneal fibrosis in the absence of LRG1 (Figure 4).

[0775] Example 3: LRG1 promotes the transformation of retinal vascular pericytes

[0776] Perivascular cells, such as pericytes contribute to ocular fibrosis. Once activated, pericytes will become proliferative, and migratory and start to secrete extracellular matrix (ECM) proteins. To understand LRG1 ’s role in pericyte activation, human retinal vascular pericytes (HRPC) were treated with recombinant human LRG1 (rhLRGI ). Western blot analysis showed that rhLRGI promotes the expression of ECM components, including connective tissue growth factor (CTGF), fibronectin, and alpha-smooth muscle actin (ASMA), as well as proteins involved in the transformation of pericytes, including N-Cadherin (N-Cad) and Plasminogen activator inhibitor-1 (PAI-1 ) (Figure 7A).

[0777] Immunofluorescence staining with a cell proliferation marker, Ki67, was used to determine proliferating pericytes following rhLRGI treatment. Results showed that rhLRGI significantly promotes HRPC proliferation (Figure 6C). A transwell assay was used to evaluate the impact of rhLRGI on HRPC migration. Results showed HRPC treated with rhLRGI are more motile (Figure 6E). Overall, the data show that LRG1 promotes HRPC activation. Example 4: The LRC region of LRG1 is responsible for its pro-fibrotic effect.

[0778] To further understand the role of LRG1 in pericyte activation and ocular fibrosis, truncated LRG1 proteins were generated containing the leucine-rich-repeat (LRR) or leucine-rich C terminal (LRRCT) domain of LRG1 . Western blot analysis showed that LRC, but not LRR, is able to promote basal and TGFpl- induced CTGF expression in HRPCs (Figure 5).

[0779] Example 5: LRG1 promotes transdifferentiation of human retinal pigment epithelium

[0780] Retinal pigment epithelial (RPE) cells maintain the health and functional integrity of both photoreceptors and the choroidal vasculature. During the development of subretinal fibrosis in nAMD, RPE lose their epithelial-like characteristics cells and subsequently undergo epithelial-mesenchymal transition (EMT) and differentiate into 9s, with increased migratory, proliferative and ECM synthesis capacity.

[0781] Assays were performed in which RPE cells were exposed to rhLRGI . Results show that rhLRGI significantly promotes RPE survival (Figure 6A), proliferation (Figure 6B), migration (Figure 6D), ECM gene transcripts (including fibronectin; FN, Connective tissue growth factor; CTGF, Transforming growth factor beta; TGFB and alpha-1 type I Collagen; COL1 a1 ) (Figure 7B), as well as fibronectin protein accumulation (Figure 7C). Western blot studies revealed that rhLRGI promotes transdifferentiation of RPE through activation of the TGFB-Smad2 / 3 signalling, and showed abolishment of Smad2 / 3 phosphorylation using SB431542, a selective inhibitor of the ALK5 / Smad2 / 3 signalling cascade (Figure 7D). This demonstrates a potential mode of action of LRG1 -mediated RPE activation.

[0782] Example 6: Identification of a panel of monoclonal chimeric antibodies with strong binding activities against distinct epitope of LRG1 and with cross-species reactivity

[0783] Having established a promoting role of LRG1 in HRPC activation, LRG1 blocking antibodies were generated using mouse immunisation followed by hybridoma and single B-cell cloning antibody discovery campaigns. More than 3,000 clones were screened and a panel of monoclonal antibodies against LRG1 was identified. Selected antibodies were further expressed as recombinant chimeric antibodies and characterised for their binding EC50 to LRG1 protein of different species as well as the LRC region (LRRCT) of LRG1 by ELISA assay. In total six chimeric anti-LRG1 antibodies showed good binding EC50 (sub-nM to double digit pM) to human and cynomolgus monkey (Macaca fascicularis) LRG1 (Table 1 ). Magacizumab (MGZ), a reported antibody against LRG-1 (WO2016 / 135462AI) shows similar binding EC50 to human LRG1 but demonstrated weaker binding activity towards monkey LRG1 and no binding activity against LRC region.

[0784] Table 1. Binding profiles of chimeric anti-LRG1 antibodies to full length of human and monkey LRG1 protein as well as the LRRCT region of human LRG1 .

[0785] The mouse species cross-reactivity was examined by Western blot in mouse retina and liver extracts. Immunoblot shows all antibodies detect LRG-1 protein band (~50kDa) in mouse liver extracts, with EBC- 59 showing the highest sensitivity in mouse retina and liver extracts (Figure 8). MGZ exhibited the weakest immunostaining against...

Claims

1. Claims:1 . An antigen-binding molecule, optionally isolated, which binds to LRG1 , wherein the antigen-binding molecule contacts the LRRCT region of LRG1 .

2. The antigen-binding molecule according to claim 1 , wherein the antigen-binding molecule contacts the region of LRG1 shown in SEQ ID NO:183, 190 or 191 .

3. The antigen-binding molecule according to claim 1 or claim 2, wherein the antigen-binding molecule is capable of inhibiting one or more functions of LRG1 .

4. The antigen-binding molecule according to any one of claims 1 to 3, wherein the antigen-binding molecule comprises:(a)(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(b)(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(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:32 HC-CDR3 having the amino acid sequence of SEQ ID NO:33; and(ii) a light chain variable (VL) region incorporating the following CDRs:LC-CDR1 having the amino acid sequence of SEQ ID NO:37 LC-CDR2 having the amino acid sequence of SEQ ID NO:26 LC-CDR3 having the amino acid sequence of SEQ ID NO:38; or(d)(i) a heavy chain variable (VH) region incorporating the following CDRs:HC-CDR1 having the amino acid sequence of SEQ ID NO:43 HC-CDR2 having the amino acid sequence of SEQ ID NO:44 HC-CDR3 having the amino acid sequence of SEQ ID NO:45; 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:48.

5. The antigen-binding molecule according to any one of claims 1 to 4, wherein the antigen-binding molecule comprises: a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:1 , 17, 31 , 42, 82, 103, 105, 107, 122, or 125; 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, 24, 36, 47, 86, 91 , 95, 98, 112, 114, 118, or 129.

6. The antigen-binding molecule according to any one of claims 1 to 5, wherein the antigen-binding molecule 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: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(ii) 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(iii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:31 ; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:36; 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:42; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:47; 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: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:86; or(vi) 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:91 ; 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: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:95; or(viii) 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:98; or(ix) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:86; or(x) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:91 ; or(xi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:95; 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:103; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:98; or(xiii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:86; 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:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:91 ; or(xv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:95; or(xvi) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NQ:105; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:98; or(xvii) 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:112; or(xviii) 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(xix) 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:118; 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:122; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:112; 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:122; 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(xxii) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:122; anda VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:118; 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:125; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:112; or(xxiv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:125; 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(xxv) a VH region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:125; and a VL region comprising an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO:118; or(xxvi) 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:129.

7. The antigen-binding molecule according to any one of claims 1 to 6, wherein the antigen-binding molecule binds to human LRG1 , mouse LRG1 , and / or cynomolgus monkey LRG1 .

8. The antigen-binding molecule according to any one of claims 1 to 7, wherein the antigen-binding molecule is a multispecific antigen-binding molecule.

9. The antigen-binding molecule according to any one of claims 1 to 8, wherein the antigen-binding molecule is conjugated to a drug moiety or a detectable moiety.

10. A chimeric antigen receptor (CAR) comprising an antigen-binding molecule according to any one of claims 1 to 8.11 . A nucleic acid, or a plurality of nucleic acids, optionally isolated, encoding the antigen-binding molecule according to any one of claims 1 to 8.

12. An expression vector, or a plurality of expression vectors, comprising a nucleic acid or a plurality of nucleic acids according to claim 11 .

13. A cell comprising an antigen-binding molecule according to any one of claims 1 to 8, a CAR according to claim 10, a nucleic acid or a plurality of nucleic acids according to claim 11 , or an expression vector or a plurality of expression vectors according to claim 12.

14. A method comprising culturing a cell according to claim 13 under conditions suitable for expression of an antigen-binding molecule or CAR by the cell.

15. A composition comprising the antigen-binding molecule according to any one of claims 1 to 9, a CAR according to claim 10, a nucleic acid or a plurality of nucleic acids according to claim 11 , an expression vector or a plurality of expression vectors according to claim 12, or a cell according to claim 13, and a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

16. An antigen-binding molecule according to any one of claims 1 to 9, a CAR according to claim 10, a nucleic acid or a plurality of nucleic acids according to claim 11 , an expression vector or a plurality of expression vectors according to claim 12, a cell according to claim 13, or composition according to claim 15, for use as a medicament.

17. An antigen-binding molecule according to any one of claims 1 to 9, a CAR according to claim 10, a nucleic acid or a plurality of nucleic acids according to claim 11 , an expression vector or a plurality of expression vectors according to claim 12, a cell according to claim 13, or composition according to claim 15, for use in the treatment or prevention of a disease or condition in which LRG1 is pathologically- implicated.

18. Use of an antigen-binding according to any one of claims 1 to 9, a CAR according to claim 10, a nucleic acid or a plurality of nucleic acids according to claim 11 , an expression vector or a plurality of expression vectors according to claim 12, a cell according to claim 13, or composition according to claim 15, in the manufacture of a medicament for use in the treatment or prevention of a disease or condition in which LRG1 is pathologically-implicated.

19. A method of treating or preventing a disease or condition in which LRG1 is pathologically-implicated, comprising administering to a subject a therapeutically- or prophylactically-effective amount of an antigenbinding according to any one of claims 1 to 9, a CAR according to claim 10, a nucleic acid or a plurality of nucleic acids according to claim 11 , an expression vector or a plurality of expression vectors according to claim 12, a cell according to claim 13, or composition according to claim 15.

20. An antigen-binding molecule, CAR, nucleic acid, expression vector, cell or composition for use according to claim 17, the use according to claim 18, or the method according to claim 19, wherein the disease or condition is characterised by one or more of: fibrosis, inflammation and pathological angiogenesis.21 . An antigen-binding molecule, CAR, nucleic acid, expression vector, cell or composition for use according to claim 17 or 20, the use according to claim 18 or 20, or the method according to claim 19 or20, wherein the disease or condition is characterised by: (i) fibrosis, inflammation and / or pathological angiogenesis of the eye; (ii) fibrosis, inflammation and / or pathological angiogenesis of the pancreas; (iii) fibrosis and / or inflammation of the joints; (iv) fibrosis and / or inflammation of the skin; (v) fibrosis and / or inflammation of the bowel; or (vi) fibrosis and / or inflammation of the kidney; (vii) fibrosis and / or inflammation of the lungs.

22. An antigen-binding molecule, CAR, nucleic acid, expression vector, cell or composition for use according to any one of claims 17, 20 or 21 , the use according to any one of claims 18, 20 or 21 , or the method according to any one of claims 19 to 21 , wherein the disease or condition is selected from: macular degeneration, Age-related Macular Degeneration (AMD), Geographic Atrophy (‘dry’ or nonexudative AMD), early AMD, early onset macular degeneration (EOMD), intermediate AMD, late / advanced AMD, ‘wet’ (neovascular or exudative) AMD, choroidal neovascularisation (CNV), retinal dystrophy, glaucoma (open-angle or closed-angle), neuromyelitis optica (neuromyelitis optica spectrum disorder (NMOSD)), diabetic macular edema (DME), diabetic retinopathy, proliferative diabetic retinopathy (PDR), polypoidal choroidal vasculopathy, proliferative vitreoretinopathy (PVR), macular edema, drusen formation, Grave's ophthalmopathy, corneal opacification, subretinal fibrosis, corneal fibrosis, epiretinal fibrosis, post-surgical fibrosis (e.g. of the posterior capsule following cataract surgery, or the bleb following trabeculectomy for glaucoma), conjunctival fibrosis, subconjunctival fibrosis, macular fibrosis preretinal fibrosis, interlobular fibrosis, periductal fibrosis, diffuse interlobular fibrosis, diffuse intralobular fibrosis, pancreatitis, cystic fibrosis, pancreatic cancer, skin fibrosis, kidney fibrosis, pulmonary fibrosis, rheumatoid arthritis, inflammatory bowel disease, diabetic nephropathy, or a diabetic wound.

23. An antigen-binding molecule, CAR, nucleic acid, expression vector, cell or composition for use according to any one of claims 17, 20 or 21 , the use according to any one of claims 18, 20 or 21 , or the method according to any one of claims 19 to 21 , wherein the disease or condition is cancer, optionally wherein the cancer is selected from: a cancer comprising cells expressing / overexpressing LRG1 , a solid tumor, a metastatic tumor, pancreatic cancer, pancreatic adenocarcinoma, pancreatic ductal adenocarcinoma, liver cancer, hepatocellular carcinoma, skin cancer, melanoma, lung cancer, lung adenocarcinoma, squamous cell lung carcinoma, non-small cell lung cancer, breast cancer, breast carcinoma, ductal carcinoma, colorectal cancer, colorectal carcinoma, colorectal adenocarcinoma, gastric cancer, gastric carcinoma, gastric adenocarcinoma, head and neck cancer, squamous cell carcinoma of the head and neck (SCCHN), ovarian cancer, ovarian carcinoma, ovarian serous adenocarcinoma, kidney cancer, renal cell carcinoma, renal clear cell carcinoma, renal cell adenocarcinoma, renal papillary cell carcinoma, cervical cancer, cervical squamous cell carcinoma, esophageal cancer, esophageal adenocarcinoma, cholangiocarcinoma, uterine cancer, uterine corpus endometrial carcinoma, thyroid cancer, thyroid carcinoma, pheochromocytoma, paraganglioma, bladder cancer, bladder urothelial carcinoma, prostate cancer, prostate adenocarcinoma, sarcoma and thymoma.

24. An in vitro complex, optionally isolated, comprising an antigen-binding molecule according to any one of claims 1 to 9 bound to LRG1 .

25. A method for detecting LRG1 in a sample, comprising contacting a sample containing, or suspected to contain, LRG1 with an antigen-binding molecule according to any one of claims 1 to 9, and detecting the formation of a complex of the antigen-binding molecule with LRG1 .

26. A method of selecting or stratifying a subject for treatment with an LRG1 -targeted agent, the method comprising contacting, in vitro, a sample from the subject with an antigen-binding molecule according to any one of claims 1 to 9 and detecting the formation of a complex of the antigen-binding molecule with LRG1.

27. Use of an antigen-binding molecule according to any one of claims 1 to 9 as an in vitro or in vivo diagnostic or prognostic agent.

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